INTRODUCTION

1. Technical Field

This invention relates to computational methods for designing ligands that bind to nuclear receptors, crystals of nuclear receptors, synthetic ligands of nuclear receptors and methods of using synthetic ligands.

2. Background

Nuclear receptors represent a superfamily of proteins that specifically bind a physiologically relevant small molecule, such as hormone or vitamin. As a result of a molecule binding to a nuclear receptor, the nuclear receptor changes the abilityof a cell to transcribe DNA, i.e. nuclear receptors modulate the transcription of DNA, although they may have transcription independent actions. Unlike integral membrane receptors and membrane associated receptors, the nuclear receptors reside in eitherthe cytoplasm or nucleus of eukaryotic cells. Thus, nuclear receptors comprise a class of intracellular, soluble ligand-regulated transcription factors.

Nuclear receptors include receptors for glucocorticoids (GRs), androgens (ARs), mineralocorticoids (MRs), progestins (PRs), estrogens (ERs), thyroid hormones (TRs), vitamin D (VDRs), retinoids (RARs and RXRs), peroxisomes (XPARs and PPARs) andicosanoids (IRs). The so called "orphan receptors" are also part of the nuclear receptor superfamily, as they are structurally homologous to the classic nuclear receptors, such as steroid and thyroid receptors. To date, ligands have not been identifiedwith orphan receptors but it is likely that small molecule ligands will be discovered in the near future for this class of transcription factors. Generally, nuclear receptors specifically bind physiologically relevant small molecules with high affinityand apparent Kd's are commonly in the 0.01-20 nM range, depending on the nuclear receptor/ligand pair.

Development of synthetic ligands that specifically bind to nuclear receptors has been largely guided by the trial and error method of drug design despite the importance of nuclear receptors in a myriad of physiological processes and medicalconditions such as hypertension, inflammation, hormone dependent cancers (e.g. breast and prostate cancer), modulation of reproductive organ function, hyperthyroidism, hypercholesterolemia and obesity. Previously, new ligands specific for nuclearreceptors were discovered in the absence of information on the three dimensional structure of a nuclear receptor with a bound ligand. Before the present invention, researchers were essentially discovering nuclear receptor ligands by probing in the darkand without the ability to visualize how the amino acids of a nuclear receptor held a ligand in its grasp.

Consequently, it would be advantageous to devise methods and compositions for reducing the time required to discover ligands to nuclear receptors, synthesize such compounds and administer such compounds to organisms to modulate physiologicalprocesses regulated by nuclear receptors.

SUMMARY OF THE INVENTION

The present invention provides for crystals of nuclear receptor ligand binding domains with a ligand bound to the ligand binding domain (LBD). The crystals of the present invention provide excellent atomic resolution of the amino acids thatinteract with nuclear receptor ligand, especially thyroid receptor ligands. The three dimensional model of a nuclear receptor LBD with a ligand bound reveals a previously unknown structure for nuclear receptors and shows that the ligand is bound in awater inaccessible binding cavity of the ligand binding domain of the nuclear receptor.

The present invention also provides for computational methods using three dimensional models of nuclear receptors that are based on crystals of nuclear receptor LBDs. Generally, the computational method of designing a nuclear receptor liganddetermines which amino acid or amino acids of a nuclear receptor LBD interact with a chemical moiety (at least one) of the ligand using a three dimensional model of a crystallized protein comprising a nuclear receptor LBD with a bound ligand, andselecting a chemical modification (at least one) of the chemical moiety to produce a second chemical moiety with a structure that either decreases or increases an interaction between the interacting amino acid and the second chemical moiety compared tothe interaction between the interacting amino acid and the corresponding chemical moiety on the natural hormone.

Also provided is a method of modulating the activity of a nuclear receptor. The method can be in vitro or in vivo. The method comprises administering in vitro or in vivo a sufficient amount of a compound of the following formula: ##STR1##

where the compound fits specially and preferentially into a nuclear hormone receptor LBD of interest. The method is exemplified by modulating the activity of a thyroid receptor (TR). For modulating TR activity, a compound of Formula I isemployed that fits spacially and preferentially into a TR ligand binding domain (TR LBD), including compounds specific for a TR LBD isoform of interest. Of particular interest are the TR LBD isoforms .alpha. (TR-.alpha.) and .beta. (TR-.beta.). Additional compounds of interest include derivatives of Formula I, such as those compounds having the biphenyl (.phi.-X-.phi.) or single phenyl (.phi.-X or X-.phi.) nucleus of Formula I and its corresponding substituent groups described herein. Compounds that are interactively designed using structural information gleaned from these compounds and which modulate nuclear hormone receptor activity also are of interest.

The present invention also includes a method for identifying a compound capable of selectively modulating the activity of a nuclear receptor. This aspect of the invention is exemplified by a method for identifying a compound capable ofselectively modulating the activity of a TR isoform. The method comprises modeling test compounds that fit spacially and preferentially into a TR LBD isoform of interest using an atomic structural model of a TR LBD isoform bound to a test compound,screening the test compounds in a biological assay for TR isoform activity characterized by binding of a test compound to a TR LBD isoform, and identifying a test compound that selectively modulates the activity of a TR isoform. The compounds may bethose of Formula I or derivatives thereof, including compounds having a biphenyl or single phenyl nucleus of Formula I.

Further included is a method for identifying agonist or antagonist ligands of a nuclear receptor using the atomic coordinates of a LBD in conjunction with a computerized modeling system. This aspect of the invention is exemplified by identifyinga TR agonist or antagonist ligand by providing the atomic coordinates of a TR LBD to a computerized modeling system, modeling ligands which fit spacially into the TR LBD, and identifying in a biological assay for TR activity a ligand which increases ordecreases TR activity. The compounds can be those of Formula I or derivatives thereof, including compounds having a biphenyl or single phenyl nucleus of Formula I.

Also provided is a method of identifying a compound that selectively modulates the activity of one type of nuclear receptor compared to other nuclear hormone receptors. The method is exemplified by modeling test compounds which fit spaciallyinto a TR LBD using an atomic structural model of a TR LBD, selecting a compound comprising conformationally constrained structural features that interact with conformationally constrained residues of a TR LBD, and identifying in a biological assay forTR activity a compound that selectively binds to a TR LBD compared to other nuclear receptors. The conformationally constrained features involved in receptor-selective ligand binding can be identified by comparing atomic models of receptor isoformsbound to the same and/or different ligands. The methods facilitate design and selection of compounds that have increased selectivity for a particular nuclear receptor. The compounds may be those of Formula I or derivatives thereof, including compoundshaving the biphenyl or single phenyl nucleus of Formula I.

Another aspect of the invention is a method for increasing the receptor selectivity of a compound for a particular type of nuclear receptor. This involves the chemical modification of a substituent group of a compound of Formula I to generatecompounds which have increased selectivity for one type of receptor. For example, chemical modification of a substituent group of the compound of Formula I can be used to introduce additional constraints into a compound that modulates TR activity toincrease its selectivity in vivo for TR-type receptors. Additional constraints also may be added for stability. The modified groups will preferably interact with a conformationally constrained structural feature of a TR LBD that is conserved among TRisoforms. A more preferred method comprises selecting compounds having conformationally constrained groups that interact with conformationally constrained residues of a TR LBD conserved among TR isoforms. The compounds can be those of Formula I orderivatives thereof, including compounds having the biphenyl or single phenyl nucleus of Formula I.

The invention finds use in the selection and characterization of peptide, peptidomimetic or synthetic compounds identified by the methods of the invention, particularly new lead compounds useful in treating disorders related to nuclearreceptor-based deficiencies, including TR-related disorders. For TR-related disorders, the compounds and methods of the invention can be used to modulate TR activity by administering to a mammal in need thereof a sufficient amount of compound of FormulaI or derivative thereof that fits spacially and preferentially into a TR LBD.

BRIEF DESCRIPTION OF THE DRAWINGS

The file of this patent contains at least one drawing executed in color. Copies of this patent with color drawings will be provided by the Patent and Trademark Office upon request and payment of necessary fee.

FIG. 1 is a diagram illustrating computational methods for designing ligands that interact with nuclear receptors of the nuclear receptor superfamily.

FIG. 2 is a schematic representation of nuclear receptor structures, indicating regions of homology within family members and functions of the various domains.

FIGS. 3A-3R shows the aligned amino acid sequences of the ligand binding domains of several members of the nuclear receptor superfamily SEQ ID NO: 1 rTR.alpha.; SEQ ID NO: 2 hTR.alpha.; SEQ ID NO: 3 hTR.beta.; SEQ ID NO: 4 hRAR.alpha.; SEQ ID NO:5 hRAR.gamma.; SEQ ID NO: 6 hRXR.alpha.; SEQ ID NO: 7 hRXR.beta.; SEQ ID NO: 8 hPPAR.alpha.; SEQ ID NO: 9 hPPAR.beta.; SEQ ID NO: 10 hPPAR.gamma.; SEQ ID NO: 11 hVDR; SEQ ID NO: 12 hER; SEQ ID NO: 13 hGR; SEQ ID NO: 14 hPR; SEQ ID NO: 15 hMR; and SEQ IDNO: 16 hAR.

FIG. 4 is a ribbon drawing of the rat TR-.alpha. LBD with secondary structure elements labelled. The ligand is depicted as a space-filling model. Alpha helices, coil conformations, and beta strands are also shown.

FIG. 5 shows two cross-sections of a space-filling model of rat TR-.alpha. exposing the ligand tightly packed within the receptor.

FIG. 6 is a schematic of the ligand binding cavity. Residues which interact with the ligand appear approximately at the site of interaction. Hydrogen bonds are shown as dashed lines between the bonding partners; distances for each bond arelisted. Non-bonded contacts are shown as radial spokes which face toward interacting atoms.

FIG. 7 is the distribution of crystallographic temperature factors in the refined rat TR-.alpha. LBD.

FIG. 8 is a ribbon drawing of the rat TR-.alpha. LBD showing the c-terminal activation domain to ligand. Residues which comprise the c-terminal activation domain (Pro393-Phe405) are depicted as a stick representation. Hydrophobic residues,particularly Phe401and Phe405 face inwards toward the ligand. Glu403 projects outward into the solvent.

FIG. 9 is an electrostatic potential surface of the rat TR-.alpha. LBD, calculated using GRAPH.

FIG. 10 is a diagram comparing agonists and antagonists for several nuclear receptors.

FIG. 11 is the synthetic scheme for preparation of TS1, TS2, TS3, TS4 and TS5.

FIG. 12 is the synthetic scheme for preparation of TS6 and TS7.

FIG. 13 is the synthetic scheme for preparation of TS8.

FIGS. 14A-14B is the synthetic scheme for preparation of TS10.

FIG. 15 depicts the chemical structures of several TR ligands.

FIG. 16 is a graph illustrating competition assays in which T.sub.3 and Triac compete with labeled T.sub.3 for binding to human TR-.alpha. or human TR-.beta..

FIGS. 17A-17B depicts a Scatchard analysis of labelled T.sub.3 binding to TR-.alpha. and TR-.beta..

FIG. 18 is a chart showing the effect of TS-10 on the transcriptional regulation of the DR4-ALP reporter gene in the presence or absence of T3 as assayed in TRAF.alpha.1 reporter cells.

FIG. 19 is a chart showing the effect of TS-10 on the transcriptional regulation of the DR4-ALP reporter gene in the presence or absence of T3 as assayed in TRAF.beta.1 reporter cells.

FIG. 20 is a chart showing the effect of TS-10 on the transcriptional regulation of the DR4-ALP reporter gene in the presence or absence of T3 as assayed in HepG2, a liver reporter cell line.

FIG. 21 is a partial ribbon drawing of TR-.alpha. LBD with T3 in the ligand binding cavity. Selected interacting amino acids are labelled, including Ile221, Ile222 and Ser260, Ala263, Ile299 and Leu 276.

FIG. 22 is a partial ribbon drawing of TR-.alpha. LBD with T3 and Dimit superimposed in the ligand binding cavity. Interactions with Ile221, Ile222, Ala260, Ile 299 and Leu276 are labelled.

FIG. 23 is a partial ribbon drawing of TR-.alpha. LBD with T3, illustrating the three Arginine residues (Arg228, Arg262 and Arg 266 (dark stick figures)) of the polar pocket, three water molecules HOH502, HOH503 and HOH504, with hydrogen bondsindicated by dotted lines.

FIG. 24 is a partial ribbon drawing of TR-.alpha. LBD with Triac, illustrating the three Arginine residues (dark stick figures) of the polar pocket, water molecules (HOH503, HOH504 and HOH600), with hydrogen bonds indicated by dotted lines.

FIG. 25 is a partial ribbon drawing of the TR-.alpha. LBD with T3 and Triac superimposed in the ligand binding cavity. The drawing shows several interacting amino acid residues in the polar pocket that remain unchanged whether T3 or Triacoccupies the ligand binding cavity: Arg262, Asn179, HOH503 and HOH504, and Ser277. Both Arg228 and Arg 266 occupy two different positions, depending on whether T3 or Triac is bound.

FIGS. 26A and 26B are stereochemical representations of the TR-.alpha. LBD with Dimit bound.

FIG. 27 is a partial ribbon drawing of TR-.beta. LBD with GC-1 in the ligand binding cavity. Amino acids Arg282, Arg316, Arg320, Asn 331 and His435 are labelled.

FIG. 28 is a partial ribbon drawing of TR-.beta. LBD with Triac in the ligand binding cavity. Amino acids Arg282, Arg316, Arg320, Asn331 and His435 are labelled.

FIG. 29 is a partial ribbon drawing of TR-.beta.LBD with GC-1 (Blue) overlayed with TR-.alpha. LBD with Dimit (Red) in the ligand binding cavities. Amino acids Arg228, Arg262, Arg266 and Ser277 (TR-.alpha. LBD), and Arg282, Arg316, Arg320 andAsn331 (TR-.beta. LBD) are labelled.

FIG. 30 is a partial ribbon drawing of TR-.beta. LBD with Triac (Blue) overlayed with TR-.alpha. LBD with Triac (Red) in the ligand binding cavities. Amino acids Arg228, Arg262, Arg266, Ser277 and His381 (TR-.alpha. LBD), and Arg282, Arg316,Arg320 and His435 (TR-.beta. LBD) are labelled.

FIG. 31 is a graph showing competition curves comparing wildtype TR-.alpha. and TR-.beta. to a variant TR-.beta. having a single amino acid substitution in the ligand binding domain.

FIG. 32 shows atomic numbering for thyronine-like ligands.

APPENDIX 1 is an appendix of references.

APPENDIX 2 is a chart of amino acids that interact with a TR ligand, for TR complexed with Dimit, Triac, IpBr2, T3 and GC-1.

APPENDIX 3 is a chart of atomic coordinates for the crystal of rat TR-.alpha. LBD complexed with Dimit.

APPENDIX 4 is a chart of atomic coordinates for the crystal of rat TR-.alpha. LBD complexed with Triac.

APPENDIX 5 is a chart of atomic coordinates for the crystal of rat TR-.alpha. LBD complexed with IpBr.sub.2.

APPENDIX 6 is a chart of atomic coordinates for the crystal of rat TR-.alpha. LBD complexed with T.sub.3.

APPENDIX 7 is a chart of atomic coordinates for the crystal of human TR-.beta. LBD complexed with Triac.

APPENDIX 8 is a chart of atomic coordinates for the crystal of human TR-.beta.-LBD complexed with GC-1.

DETAILED DESCRIPTION OF THE INVENTION

INTRODUCTION

The present invention provides new methods, particularly computational methods, and compositions for the generation of nuclear receptor synthetic ligands based on the three dimensional structure of nuclear receptors, particularly the thyroidreceptor (herein referred to as "TR"). Previously, the lack of three dimensional structural information about the ligand binding domain of a nuclear receptor thwarted the field of nuclear receptor drug discovery, especially the absence of threedimensional structural information relating to a nuclear receptor with a ligand bound.

Described herein for the first time are crystals and three dimensional structural information from a nuclear receptor's ligand binding domain (LBD) with a ligand bound. The structure of the TR LBD complexed with 3,5,3'-triiodothyronine(T.sub.3), 3,5-dibromo-3'-isopropylthyronine (IpBr.sub.2), 3,5-dimethyl-3'-isopropylthyronine (Dimit), and 3,5,3'-triiodothyroacetic acid (Triac), 3,5-dimethyl-4-(4'-hydroxy-3'isopropylbenzyl)-phenoxy acetic acid (GC1) are exemplified. Such crystalsoffer superior resolution at the atomic level and the ability to visualize the coordination of nuclear receptor ligands by amino acids that comprise the LBD. The present invention also provides computational methods for designing nuclear receptorsynthetic ligands using such crystal and three dimensional structural information to generate synthetic ligands that modulate the conformational changes of a nuclear receptor's LBD. Such synthetic ligands can be designed using the computational methodsdescribed herein and shown, in part, in FIG. 1. These computational methods are particularly useful in designing an antagonist or partial agonist to a nuclear receptor, wherein the antagonist or partial agonist has an extended moiety that prevents anyone of a number of ligand-induced molecular events that alter the receptor's influence on the regulation of gene expression, such as preventing the normal coordination of the activation domain observed for a naturally occurring ligand or other ligandsthat mimic the naturally occurring ligand, such as an agonist. As described herein, synthetic ligands of nuclear receptors will be useful in modulating nuclear receptor activity in a variety of medical conditions.

Of particular interest is use of such ligands in a method of modulating TR activity in a mammal by administering to a mammal in need thereof a sufficient amount of a compound of Formula I, ##STR2##

where the compound fits spatially and preferentially into a TR LBD. By "fits spacially" is intended that the three-dimensional structure of a compound is accommodated geometrically by a cavity or pocket of a TR LBD. By "TR LBD" is intended astructural segment or segments of thyroid hormone receptor polypeptide chain folded in such a way so as to give the proper geometry and amino acid residue configuration for ligand binding. This is the physical arrangement of protein atoms inthree-dimensional space forming a ligand binding pocket or cavity. By "fits spacially and preferentially" is intended that a compound possesses a three-dimensional structure and conformation for selectively interacting with a TR LBD. Compounds ofinterest also include derivatives of Formula I. By "derivatives of Formula I" is intended compounds that comprise at least a single phenyl scaffold (.phi.-X or X-.phi.) of the biphenyl scaffold (.phi.-X-.phi.) of Formula I which comprise thecorresponding substituents of Formula I described herein. Compounds that are interactively designed using structural information gleaned from these compounds and which modulate nuclear hormone receptor activity also are of interest. Preferred compoundsof Formula I and its derivatives that fit spacially and preferentially into a TR LBD comprise the following substituents:

(i) an R1-substituent comprising an anionic group that interacts with a side chain nitrogen atom of an arginine corresponding to a residue from the group Arg228, Arg262, and Arg266 of human TR-.alpha., and Arg282, Arg316 and Arg320 of humanTR-.beta., where the anionic group is about 1.7-4.0 .ANG. from the nitrogen atom;

(ii) an R2-substituent comprising a hydrophobic or hydrophilic group that fits spacially into the TR LBD;

(iii) an R3-substituent comprising a hydrophobic or hydrophilic group that interacts with a side chain atom of a serine, alanine and/or isoleucine corresponding to a residue from the group Ser260, Ala263 and Ile299 of human TR-.alpha., andSer314, Ala317 and Ile352 of human TR-.beta., where the hydrophobic or hydrophilic group is about 1.7-4.0 .ANG. from the side chain atom;

(iv) an R5-substituent comprising a hydrophobic or hydrophilic group that interacts with a side chain atom of a phenylalanine and/or isoleucine corresponding to a residue from the group Phe218, Ile221 and Ile222 of human TR-.alpha., and Phe272,Ile275 and Ile276 of human TR-.beta., where the hydrophobic or hydrophilic group is about 1.7-4.0 .ANG. from the side chain atom;

(v) an R6-substituent comprising a hydrophobic or hydrophilic group that fits spacially into the TR LBD;

(vi) an X-substituent comprising a hydrophobic or hydrophilic group that interacts with a side chain atom of a leucine corresponding to a residue from the group Leu276 and Leu292 of human TR-.alpha., and Leu 330 and Leu346 of human TR-.beta.,where the hydrophobic or hydrophilic group is about 1.7-4.0 .ANG. from the side chain atom;

(vii) an R2'-substituent comprising a hydrophobic or hydrophilic group that fits spacially into the TR LBD;

(viii) an R3'-substituent comprising a hydrophobic group that interacts with a side chain atom of a phenylalanine, glycine and/or methionine corresponding to a residue from the group Phe215, Gly290, and Met388 of human TR-.alpha., and Phe269,Gly344, Met442 of human TR-.beta., where the hydrophobic group is about 1.7-4.0 .ANG. from the side chain atom;

(ix) an R4'-substituent comprising an hydrogen bond donor or acceptor group that interacts with a side chain carbon or nitrogen atom of a histidine corresponding to residue His381 of human TR-.alpha., and His435 of human TR-.beta., where thehydrogen bond donor or acceptor group is about 1.7-4.0 .ANG. from the side chain atom;

(x) an R5'-substituent comprising a hydrophobic or hydrophilic group that fits spacially into the TR LBD;

(xi) and R6'-substituent comprising a hydrophobic or hydrophilic group that fits spacially into the TR LBD; and

where the compound is other than thyronine (T3), triiodothyronine (T4) or other thyronine-like compounds previously known and used in a TR treatment method, such as those referenced in Appendix I.

Examples of such substituents include the following:

where R.sub.1 is

--O--CH.sub.2 CO.sub.2 H, --NHCH.sub.2 CO.sub.2 H, --CO.sub.2 H, --CH.sub.2 CO.sub.2 H, --CH.sub.2 CH.sub.2 CO.sub.2 H, --CH.sub.2 CH.sub.2 CH.sub.2 CO.sub.2 H, --CH.sub.2 CH(NH.sub.2)CO.sub.2 H, --CH.sub.2 CH[NHCOCH.phi..sub.2 ]CO.sub.2 H,--CH.sub.2 CH[NHCO(CH.sub.2).sub.15 CH.sub.3 ]CO.sub.2 H, --CH.sub.2 CH[NH--FMOC]CO.sub.2 H, --CH.sub.2 CH[NH-tBOC]CO.sub.2 H, or a carboxylate connected to the ring with a 0 to 3 carbon linker, --PO.sub.3 H.sub.2, --CH.sub.2 PO.sub.3 H.sub.2, --CH.sub.2CH.sub.2 PO.sub.3 H.sub.2, --CH.sub.2 CHNH.sub.2 PO.sub.3 H.sub.2, --CH.sub.2 CH[NHCOCH.phi..sub.2 ]PO.sub.3 H.sub.2, --CH.sub.2 CH[NHCO(CH.sub.2).sub.15 CH.sub.3 ]PO.sub.3 H.sub.2, --CH.sub.2 CH[NH-FMOC]PO.sub.3 H.sub.2, --CH.sub.2 CH[NH-tBOC]PO.sub.3H.sub.2, or a phosphate or phosphonate connected to the ring with a 0 to 3 carbon linker, --SO.sub.3 H, --CH.sub.2 SO.sub.3 H, --CH.sub.2 CH.sub.2 SO.sub.3 H, --CH.sub.2 CHNH.sub.2 SO.sub.3 H, --CH.sub.2 CH[NHCOCH.phi..sub.2 ]SO.sub.3 H, --CH.sub.2CH[NHCO(CH.sub.2).sub.15 CH.sub.3 ]SO.sub.3 H, --CH.sub.2 CH[NH-FMOC]SO.sub.3 H, --CH.sub.2 CH[NH-tBOC]SO3H, or a sulfate or sulfite connected to the ring with a 0 to 3 carbon linker, or acts as the functional equivalent of CH.sub.2 CH(NH.sub.2)CO.sub.2H of T3 in the molecular recognition domain when bound to a TR, wherein R.sub.1 can be optionally substituted with an amine,

where R.sub.2 is

H, halogen, CF.sub.3, OH, NH.sub.2, SH, CH.sub.3, --Et, or acts as the functional equivalent of H in the molecular recognition domain when bound to a TR,

where R.sub.3 is

--H, halogen, --CF.sub.3, --OH, --NH.sub.2, --N.sub.3, --SH, --CH.sub.3, --Et, or acts as the functional equivalent of I in the molecular recognition domain when bound to a TR,

where R.sub.5 is

--H, halogen, --CF.sub.3, --OH, --NH.sub.2, --N.sub.3, --SH, --CH.sub.3, --Et, or acts as the functional equivalent of I in the molecular recognition domain when bound to a TR, and R.sub.3 can be identical to R.sub.5,

where R.sub.6 is

--H, halogen, --CF.sub.3, --OH, --NH.sub.2, --SH, --CH.sub.3, or acts as the functional equivalent of H in the molecular recognition domain when bound to a TR, and R.sub.2 can be identical to R.sub.6,

where R.sub.2 ' is

--H, halogen, --CF.sub.3, --OH, --NH.sub.2, --N.sub.3, --SH, --CH.sub.3, --Et, or acts as the functional equivalent of H in the molecular recognition domain when bound to a TR,

where R.sub.3 ' is any hydrophobic group, including

halogen, --CF.sub.3, --SH, alkyl, aryl, 5- or 6-membered heterocyclie, cyano, or acts as the functional equivalent of I in the molecular recognition domain when bound to a TR,

where R.sub.4 ' is

--H, halogen, --CF.sub.3, --OH, --NH.sub.2, NH.sub.3, --N(CH.sub.3).sub.3, carboxylate, phosphonate, phosphate or sulfate, --SH, --CH.sub.3, --Et, or akyl, aryl or 5- or 6-membered heterocyclic aromatic attached through urea or carbamate linkagesto O or N or S at the R.sub.4 ' position, or acts as the functional equivalent of OH in the molecular recognition domain when bound to a TR,

where R.sub.5 ' is

--H, --OH, --NH.sub.2, --N(CH.sub.3).sub.2 --SH --NH.sub.3, --N(CH.sub.3).sub.3, carboxylate, phosphonate, phosphate, sulfate, branched or straight chain alkyl having 1 to 9 carbons, substituted or unsubstituted aryl, wherein said substitutedaryl is substituted with halogen or 1 to 5 carbon alkyl and wherein said aryl is optionally connected to the ring by a --CH.sub.2 --, aromatic heterocycle having 5 to 6 atoms, wherein said heterocycle may be substituted with one or more groups selectedfrom --OH, --NH.sub.2, --SH, --NH.sub.3, --N(CH.sub.3).sub.3, carboxylate, phosphonate, phosphate or sulfate, heteroalkyl, arylalkyl, heteroaryl alkyl, polyaromatic, or polyheteroaromatic, wherein said R.sub.5 ' may be substituted with polar or chargedgroups,

where R.sub.6 ' is

--H, halogen, --CF.sub.3, --OH, --NH.sub.2, --SH, --CH.sub.3, --Et, or acts as the functional equivalent of H in the molecular recognition domain when bound to a TR,

where X is

O, S, SO.sub.2, NH, NR.sub.7, CH.sub.2, CHR.sub.7, CR.sub.7 R.sub.7, wherein R.sub.7 is alkyl, aryl or 5- or 6-membered heterocyclic aromatic,

and where the TR LBD ligand has an apparent Kd for binding TR LBD of 1 .mu.M or less.

Of particular interest are the class of compounds according to Formula I having the following substituents: where R.sub.1 is carboxylate, phosphonate, phosphate or sulfite and is connected to the ring with a 0 to 3 carbon linker, R.sub.2 is H,R.sub.3 is --I, --Br, or --CH.sub.3, R.sub.5 is --I, --Br, or --CH.sub.3, R.sub.6 is H, R.sub.2 ' is H, R.sub.3 ' is --I, --Br, --CH.sub.3, --iPr, -phenyl, benzyl, or 5- or 6-membered ring heterocycles, R.sub.4 ' is --OH, --NH.sub.2, and --SH, R.sub.5 'is --H, --OH, --NH.sub.2, --N(CH.sub.3).sub.2 --SH--NH.sub.3, --N(CH.sub.3).sub.3, carboxylate, phosphonate, phosphate, sulfate, branched or straight chain alkyl having 1 to 9 carbons, substituted or unsubstituted aryl, wherein said substituted aryl issubstituted with halogen or 1 to 5 carbon alkyl and wherein said aryl is optionally connected to the ring by a --CH.sub.2 --, aromatic heterocycle having 5 to 6 atoms, wherein said heterocycle may be substituted with one or more groups selected from--OH, --NH.sub.2, --SH, --NH.sub.3, --N(CH.sub.3).sub.3, carboxylate, phosphonate, phosphate or sulfate, heteroalkyl, arylalkyl, heteroaryl alkyl, polyaromatic, or polyheteroaromatic, wherein said R.sub.5 ' may be substituted with polar or chargedgroups, and R.sub.6 ' is H.

The present invention also includes a method for identifying a compound capable of selectively modulating the activity of a TR isoform. By "modulating" is intended increasing or decreasing activity of a TR. By "TR isoform" is intended TRproteins encoded by subtype and variant TR genes. This includes TR-.alpha. and TR-.beta. isoforms encoded by different genes (e.g., thra and thrb) and variants of the same genes (e.g., thrb1 and thrb2). The method comprises the steps of modeling testcompounds that fit spacially and preferentially into a TR LBD isoform of interest using an atomic structural model of a TR LBD isoform bound to a test compound, screening the test compounds in a biological assay for TR isoform activity characterized bybinding of a test compound to a TR LBD isoform, and identifying a test compound that selectively modulates the activity of a TR isoform. By "modeling" is intended quantitative and qualitative analysis of receptor-ligand structure/function based onthree-dimensional structural information and receptor-ligand interaction models. This includes conventional numeric-based molecular dynamic and energy minimization models, interactive computer graphic models, modified molecular mechanics models,distance geometry and other structure-based constraint models. Modeling is preferably performed using a computer and may be further optimized using known methods.

For selectively modulating activity of a TR isoform, such as TR-.alpha. or TR-.beta., a sufficient amount of a compound that fits spatially and preferentially into TR LBD isoform is provided in vitro or in vivo to achieve the desired end result. TR-.alpha. isoform selectivity can be accomplished with a compound comprising an anionic group that interacts with an oxygen or carbon of a serine residue corresponding to Ser277 of human TR-.alpha., where the anionic group is about 1.7-4.0 .ANG. fromthe side chain atom. TR-.beta. isoform selectivity can be accomplished with a compound comprising an anionic group that interacts with the side chain nitrogen of an asparagine corresponding to Asn331 of human TR-.beta., where the anionic group is about1.7-4.0 .ANG. from the side chain nitrogen atom.

The present invention further includes a method for identifying a TR agonist or antagonist ligand by providing the atomic coordinates of a TR LBD to a computerized modeling system, modeling ligands which fit spacially into the TR LBD, andidentifying in a biological assay for TR activity a ligand which increases or decreases the activity of the TR.

The invention also involves a method for increasing receptor selectivity of a compound of Formula I or derivatives thereof for a TR-type receptor versus other nuclear receptors by selecting a compound that interacts with conformationallyconstrained residues of a TR LBD that are conserved among TR isoforms. "Conformationally constrained" is intended to refer to the three-dimensional structure of a chemical or moiety thereof having certain rotations about its bonds fixed by various localgeometric and physical-chemical constraints. In designing and selecting compounds having increased specificity for TRs compared to other nuclear receptors, the following methods of the invention can be used. One method involves comparing atomic modelsof a first TR LBD isoform bound to a compound with a second TR LBD isoform bound to the same compound, identifying atoms of the TR LBD and compounds which interact, and designing or selecting a compound that interacts with TR LBD residues comprising aconformationally constrained structural feature that is conserved between the TR LBD isoforms. Another method relates to comparing a first TR LBD complexed with a first compound to a second TR LBD complexed with a second compound having one or moredifferent substituents compared to the first compound, identifying atoms of the TR LBD and compounds which interact, and designing or selecting compounds that interact with TR LBD residues comprising a conformationally constrained structural feature thatis conserved between the TR LBD isoforms. The methods also facilitate identification of structural and conformationally constrained interactions that are conserved between compounds that bind to a TR LBD. The methods are exemplified by comparing atomicmodels of a first TR LBD isoform complexed with a first compound of Formula I to a second TR LBD isoform complexed with the first compound, or a second compound of Formula I having different substituents than the first compound. For example, aTR-.alpha. LBD bound to a natural hormone such as T3 is compared to a TR-.beta. LBD bound to an organic thyronine-like compound such as GC-1. Conserved contacts are identified which are made between atoms of the different compounds and atoms of the TRLBDs, and the fiducial and adjustable components identified. Compounds selective for TR are identified in a biological assay for TR activity that assays for selective binding to a TR and/or TR LBD compared to other nuclear receptors. Conventionalassays for TR and other nuclear receptors may be conducted in parallel or serially, including those assays described herein. Automatable methods are preferred. The methods facilitate design and selection of compounds comprising cyclic carbon andsubstituent atoms that interact with a constrained side chain and/or main chain atom of a TR LBD residue.

In another aspect of the invention, the methods described herein are useful for selecting peptides, peptidomimetics or synthetic molecules that modulate TR activity. Methods of the invention also find use in characterizing structure/functionrelationships of natural and synthetic TR-ligands. Molecules of particular interest are new thyronine-like compounds other than T3, T4 and other thyronine-like compounds previously known and used for treating TR-related disorders. New compounds of theinvention include those which bind to a TR LBD isoform with greater affinity than T3 or T4 and those which exhibit isoform-specific binding affinity.

APPLICABILITY TO NUCLEAR RECEPTORS

The present invention, particularly the computational methods, can be used to design drugs for a variety of nuclear receptors, such as receptors for glucocorticoids (GRs), androgens (ARs), mineralocorticoids (MRs), progestins (PRs), estrogens(ERs), thyroid hormones (TRs), vitamin D (VDRs), retinoid (RARs and RXRs), icosanoid (IRs), and peroxisomes (XPARS and peroxisomal proliferators (PPAP)). The present invention can also be applied to the "orphan receptors," as they are structurallyhomologous in terms of modular domains and primary structure to classic nuclear receptors, such as steroid and thyroid receptors. The amino acid homologies of orphan receptors with other nuclear receptors ranges from very low (<15%) to in the rangeof 35% when compared to rat RAR.alpha. and human TR-.beta. receptors, for example. In addition, as is revealed by the X-ray crystallographic structure of the TR and structural analysis disclosed herein, the overall folding of liganded superfamilymembers is likely to be similar. Although ligands have not been identified with orphan receptors, once such ligands are identified one skilled in the art will be able to apply the present invention to the design and use of such ligands, as their overallstructural modular motif will be similar to other nuclear receptors described herein.

Modular Functional Domains of Nuclear Receptors

The present invention will usually be applicable to all nuclear receptors, as discussed herein, in part, to the patterns of nuclear receptor activation, structure and modulation that have emerged as a consequence of determining the threedimensional structures of nuclear receptors with different ligands bound, notably the three dimensional structures or crystallized protein structure of the ligand binding domains for TR-.alpha. and TR-.beta.. Proteins of the nuclear receptorsuperfamily display substantial regions of amino acid homology, as described herein and known in the art see FIG. 2. Members of this family display an overall structural motif of three modular domains (which is similar to the TR three modular domainmotif):

1) a variable amino-terminal domain;

2) a highly conserved DNA-binding domain (DBD); and

3) a less conserved carboxyl-terminal LBD. The modularity of this superfamily permits different domains of each protein to separately accomplish different functions, although the domains can influence each other. The separate function of adomain is usually preserved when a particular domain is isolated from the remainder of the protein. Using conventional protein chemistry techniques a modular domain can sometimes be separated from the parent protein. Using conventional molecularbiology techniques each domain can usually be separately expressed with its original function intact or chimerics of two different nuclear receptors can be constructed, wherein the chimerics retain the properties of the individual functional domains ofthe respective nuclear receptors from which the chimerics were generated.

FIG. 2 provides a schematic representation of family member structures, indicating regions of homology within family members and functions of the various domains.

Amino Terminal Domain

The amino terminal domain is the least conserved of the three domains and varies markedly in size among nuclear receptor superfamily members. For example, this domain contains 24 amino acids in the VDR and 603 amino acids in the MR. This domainis involved in transcriptional activation and in some cases its uniqueness may dictate selective receptor-DNA binding and activation of target genes by specific receptor isoforms. This domain can display synergistic and antagonistic interactions withthe domains of the LBD. For example, studies with mutated and/or deleted receptors show positive cooperativity of the amino and carboxy terminal domains. In some cases, deletion of either of these domains will abolish the receptor's transcriptionalactivation functions.

DNA-Binding Domain

The DBD is the most conserved structure in the nuclear receptor superfamily. It usually contains about 70 amino acids that fold into two zinc finger motifs, wherein a zinc ion coordinates four cysteines. DBDs contain two perpendicularlyoriented .alpha.-helixes that extend from the base of the first and second zinc fingers. The two zinc fingers function in concert along with non-zinc finger residues to direct nuclear receptors to specific target sites on DNA and to align receptorhomodimer or heterodimer interfaces. Various amino acids in DBD influence spacing between two half-sites (usually comprised of six nucleotides) for receptor dimer binding. For example, GR subfamily and ER homodimers bind to half-sites spaced by threenucleotides and oriented as palindromes. The optimal spacings facilitate cooperative interactions between DBDs, and D box residues are part of the dimerization interface. Other regions of the DBD facilitate DNA-protein and protein-protein interactionsrequired for RXR homodimerization and heterodimerization on direct repeat elements.

The LBD may influence the DNA binding of the DBD, and the influence can also be regulated by ligand binding. For example, TR ligand binding influences the degree to which a TR binds to DNA as a monomer or dimer. Such dimerization also dependson the spacing and orientation of the DNA half sites. The receptors also can interact with other proteins and function to regulate gene expression.

The nuclear receptor superfamily has been subdivided into two subfamilies: 1) GR (GR, AR, MR and PR) and 2) TR (TR, VDR, RAR, RXR, and most orphan receptors) on the basis of DBD structures, interactions with heat shock proteins (hsp), and abilityto form heterodimers. GR subgroup members are tightly bound by hsp in the absence of ligand, dimerize following ligand binding and dissociation of hsp, and show homology in the DNA half sites to which they bind. These half sites also tend to bearranged as palindromes. TR subgroup members tend to be bound to DNA or other chromatin molecules when unliganded, can bind to DNA as monomers and dimers, but tend to form heterodimers, and bind DNA elements with a variety of orientations and spacingsof the half sites, and also show homology with respect to the nucleotide sequences of the half sites. By this classification, ER does not belong to either subfamily, since it resembles the GR subfamily in hsp interactions, and the TR subfamily innuclear localization and DNA-binding properties.

Ligand Binding Domain

The LBD is the second most highly conserved domain in these receptors. Whereas integrity of several different LBD sub-domains is important for ligand binding, truncated molecules containing only the LBD retain normal ligand-binding activity. This domain also participates in other functions, including dimerization, nuclear translocation and transcriptional activation, as described herein. Importantly, this domain binds the ligand and undergoes ligand-induced conformational changes asdetailed herein.

Most members of the superfamily, including orphan receptors, possess at least two transcription activation subdomains, one of which is constitutive and resides in the amino terminal domain (AF-1), and the other of which (AF-2 (also referenced asTAU 4)) resides in the ligand-binding domain whose activity is regulated by binding of an agonist ligand. The function of AF-2 requires an activation domain (also called trsactivation domain) that is highly conserved among the receptor superfamily(approximately amino acids 1005 to 1022). Most LBDs contain an activation domain. Some mutations in this domain abolish AF-2 function, but leave ligand binding and other functions unaffected. Ligand binding allows the activation domain to serve as aninteraction site for essential co-activator proteins that function to stimulate (or in some cases, inhibit) transcription.

For example, Shibata, H., et al. (Recent Progress in Hormone Res. 52:141-164 (1997)) has; reviewed the role of co-activators and co-repressors in steroid/thyroid hormone receptor systems. Steroid receptor co-activator-one (SRC-1) appears to bea general co-activator for all AF-2 domain containing receptors tested. SRC-1 enhances transactivation of steroid hormone-dependent target genes. Other putative co-activators have been reported, including the SRC-1 related proteins, TIF-2 and GRIP-1,and other putative unrelated co-activators such as ARA-70, Trip 1, RIP-140, and TIF-1. In addition another co-activator CREB-binding protein (CBP) has been shown to enhance receptor-dependent target gene trascription. CBP and SRC-1 interact andsynergistically enhance transcriptional activation by the ER and PR. A ternary complex of CBP, SRC-1, and liganded receptors-may form to increase the rate of hormone-responsive gene transcription. Co-repressors, such as SMRT and N-CoR, for TR and RAR,have been identified that also contribute to the silencing function of unliganded TR. The unliganded TR and RAR have been shown to inhibit basal promoter activity; this silencing of target gene transcription by unliganded receptors is mediated by theseco-repressors. The collective data suggests that upon binding of agonist, the receptor changes its conformation in the ligand-binding domain that enables recruitment of co-activators, which allows the receptor to interact with the basal transcriptionalmachinery more efficiently and to activate transcription. In contrast, binding of antagonists induces a different conformational change in the receptor. Although some antagonist-bound receptors can dimerize and bind to their cognate DNA elements, theyfail to dislodge the associated co-repressors, which results in a nonproductive interaction with the basal transcriptional machinery. Similarly, the TR and RAR associate with co-repressors in the absence of ligand, thereby resulting in a negativeinteraction with the transcriptional machinery that silences target gene expression. In the case of mixed agonist/antagonists, such as 4-hydroxytamoxifen, activation of gene transcription may depend on the relative ratio of co-activators andco-repressors in the cell or cell-specific factors that determine the relative agonistic or antagonistic potential of different compounds. These co-activators and co-repressors appear to act as an accelerator and/or a brake that modulatestranscriptional regulation of hormone-responsive target gene expression.

The carboxy-terminal activation subdomain, as described herein is in close three dimensional proximity in the LBD to the ligand, so as to allow for ligands bound to the LBD to coordinate (or interact) with amino acid(s) in the activationsubdomain. As described herein, the LBD of a nuclear receptor can be expressed, crystallized, its three dimensional structure determined with a ligand bound (either using crystal data from the same receptor or a different receptor or a combinationthereof), and computational methods used to design ligands to its LBD, including ligands that contain an extension moiety that coordinates the activation domain of the nuclear receptor.

Once a computationally designed ligand (CDL) is synthesized as described herein and known in the art, it can be tested using assays to establish its activity as an agonist, partial agonist or antagonist, and affinity, as described herein. Aftersuch testing, the CDLs can be further refined by generating LBD crystals with a CDL bound to the LBD. The structure of the CDL can then be further refined using the chemical modification methods described herein for three dimensional models to improvethe activity or affinity of the CDL and make second generation CDLs with improved properties, such as that of a super agonist or antagonist described herein. Agonist and antagonist ligands also can be selected that modulate nuclear receptor responsivegene transcription through altering the interaction of co-activators and co-repressors with their cognate nuclear hormone receptor. For example, CDL agonists can be selected that block or dissociate the co-repressor from interaction with the receptor,and/or which promote binding or association of the co-activator. CDL antagonists can be selected that block co-activator interaction and/or promote co-repressor interaction with the target receptor. Selection can be done in binding assays that screenfor CDLs having the desired agonist or antagonist properties. Suitable assays for such screening are described herein and in Shibata, H., et al. (Recent Prog. Horm. Res. 52:141-164 (1997)); Tagami, T., et al. (Mol. Cell Biol. 17(5):2642-2648(1997)); Zhu, X G., et al. (J. Biol. Chem. 272(14):9048-9054 (1997)); Lin, B. C., et al. (Mol. Cell Biol. 17(10):6131-6138 (1997)); Kakizawa, T., et al. (J. Biol. Chem. 272(38):23799-23804 (1997)); and Chang, K. H., et al. (Proc. Natl. Acad. Sci. USA 94(17):9040-9045 (1997)), which references are incorporated herein in their entirety by reference.

NUCLEAR RECEPTOR ISOFORMS

The present invention also is applicable to generating new synthetic ligands to distinguish nuclear receptor isoforms. As described herein, CDLs can be generated that distinguish between binding isoforms, thereby allowing the generation ofeither tissue specific or function specific synthetic ligands. For instance, GR subfamily members have usually one receptor encoded by a single gene, although are exceptions. For example, there are two PR isoforms, A and B, translated from the samemRNA by alternate initiation from different AUG codons. There are two GR forms, one of which does not bind ligand. This method is especially applicable to the TR subfamily which usually has several receptors that are encoded by at least two (TR:.alpha., .beta.) or three (RAR, RXR, and PPAR: .alpha., .beta., .gamma.) genes or have alternate RNA splicing and such an example for TR is described herein.

NUCLEAR RECEPTOR CRYSTALS

The invention provides for crystals made from nuclear receptor ligand binding domains with the ligand bound to the receptor. As exemplified in the Examples, TRs are crystallized with a ligand bound to it. Crystals are made from purified nuclearreceptor LBDs that are usually expressed by a cell culture, such as E. coli. Preferably, different crystals (cc-crystals) for the same nuclear receptor are separately made using different ligands, such as a naturally occurring ligand and at least onebromo- or iodo- substituted synthetic ligand that acts as an analog or antagonist of the naturally occurring ligand. Such bromo- and iodo- substitutions act as heavy atom substitutions in nuclear receptor ligands and crystals of nuclear receptorproteins. This method has the advantage for phasing of the crystal in that it bypasses the need for obtaining traditional heavy metal derivatives. After the three dimensional structure is determined for the nuclear receptor LBD with its ligand bound,the three dimensional structure can be used in computational methods to design a synthetic ligand for the nuclear receptor and further activity structure relationships can be determined through routine testing using the assays described herein and knownin the art.

Expression and Purification of other Nuclear Receptor LBD Structures

High level expression of nuclear receptor LBDs can be obtained by the techniques described herein as well as others described in the literature. High level expression in E. coli of ligand binding domains of TR and other nuclear receptors,including members of the steroid/thyroid receptor superfamily, such as the receptors ER, AR, MR, PR, RAR, RXR and VDR can also be achieved. Yeast and other eukaryotic expression systems can be used with nuclear receptors that bind heat shock proteins asthese nuclear receptors are generally more difficult to express in bacteria, with the exception of ER, which can be expressed in bacteria. Representative nuclear receptors or their ligand binding domains have been cloned and sequenced: humanRAR-.alpha., human RAR-.gamma., human RXR-.alpha., human RXR-.beta., human PPAR-.alpha., human PPAR-.beta., human PPAR-.gamma., human VDR, human ER (as described in Seielstad et al., Molecular Endocrinology, vol 9:647-658 (1995), incorporated herein byreference), human GR, human PR, human MR, and human AR. The ligand binding domain of each of these nuclear receptors has been identified and is shown in FIGS. 3A-3R. Using the information in FIGS. 3A-3R in conjunction with the methods described hereinand known in the art, one of ordinary skill in the art could express and purify LBDs of any of the nuclear receptors, including those illustrated in FIGS. 3A-3R, bind it to an appropriate ligand, and crystallize the nuclear receptor's LBD with a boundligand.

FIGS. 3A-3R is an alignment of several members of the steroid/thyroid hormone receptor superfamily that indicates the amino acids to be included in a suitable expression vector.

Extracts of expressing cells are a suitable source of receptor for purification and preparation of crystals of the chosen receptor. To obtain such expression, a vector is constructed in a manner similar to that employed for expression of the ratTR alpha (Apriletti et al. Protein Expression and Purification, 6:363-370 (1995), herein incorporated by reference). The nucleotides encoding the amino acids encompassing the ligand binding domain of the receptor to be expressed, for example theestrogen receptor ligand binding domain (hER-LBD) (corresponding to R at position 725 to L at position 1025 as standardly aligned as shown in the FIG. 3), are inserted into an expression vector such as the one employed by Apriletti et al (1995). For thepurposes of obtaining material that will yield good crystals it is preferable to include at least the amino acids corresponding to human TR-.beta. positions 725 to 1025. Stretches of adjacent amino acid sequences may be included if more structuralinformation is desired. Thus, an expression vector for the human estrogen receptor can be made by inserting nucleotides encoding amino acids from position 700 to the c-terminus at position 1071. Such a vector gives high yield of receptor in E. colithat can bind hormone (Seielstad et al. Molecular Endocrinology 9:647-658 (1995)). However, the c-terminal region beyond position 1025 is subject to variable proteolysis and can advantageously be excluded from the construct, this technique of avoidingvariable proteolysis can also be applied to other nuclear receptors.

TR-.alpha. And TR-.beta. As Examples of Nuclear Receptor LBD Structure and Function TR Expression, Purification And Crystallization

As an example of nuclear receptor structure of the ligand binding domain the .alpha.- and .beta.-isoforms of TR are crystallized from proteins expressed from expression constructs, preferably constructs that can be expressed in E. coli. Otherexpression systems, such as yeast or other eukaryotic expression systems can be used. For the TR, the LBD can be expressed without any portion of the DBD or amino-terminal domain. Portions of the DBD or amino-terminus can be included if furtherstructural information with amino acids adjacent the LBD is desired. Generally, for the TR the LBD used for crystals will be less than 300 amino acids in length. Preferably, the TR LBD will be at least 150 amino acids in length, more preferably atleast 200 amino acids in length, and most preferably at least 250 amino acids in length. For example the LBD used for crystallization can comprise amino acids spanning from Met 122 to Val 410 of the rat TR-.alpha., Glu 202 to Asp 461 of the humanTR-.beta..

Typically TR LBDs are purified to homogeneity for crystallization. Purity of TR LBDs is measured with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), mass spectrometry (MS) and hydrophobic high performance liquidchromatography (HPLC). The purified TR for crystallization should be at least 97.5% pure or 97.5%, preferably at least 99.0% pure or 99.0% pure, more preferably at least 99.5% pure or 99.5% pure.

Initially purification of the unliganded receptor can be obtained by conventional techniques, such as hydrophobic interaction chromatography (HPLC), ion exchange chromatography (HPLC), and heparin affinity chromatography.

To achieve higher purification for improved crystals of nuclear receptors, especially the TR subfamily and TR, it will be desirable to ligand shift purify the nuclear receptor using a column that separates the receptor according to charge, suchas an ion exchange or hydrophobic interaction column, and then bind the eluted receptor with a ligand, especially an agonist. The ligand induces a change in the receptor's surface charge such that when re-chromatographed on the same column, the receptorthen elutes at the position of the liganded receptor are removed by the original column run with the unliganded receptor. Usually saturating concentrations of ligand are used in the column and the protein can be preincubated with the ligand prior topassing it over the column. The structural studies detailed herein indicate the general applicability of this technique for obtaining super-pure nuclear receptor LBDs for crystallization.

More recently developed methods involve engineering a "tag" such as with histidine placed on the end of the protein, such as on the amino terminus, and then using a nickle chelation column for purification, Janknecht R., Proc. Natl. Acad. Sci. USA, 88:8972-8976 (1991) incorporated by reference.

To determine the three dimensional structure of a TR LBD, or a LBD from another member of the nuclear receptor superfamily, it is desirable to co-crystalize the LBD with a corresponding LBD ligand. In the case of TR LBD, it is preferable toseparately co-crystalize it with ligands such as T3, IpBr and Dimit that differ in the heavy atoms which they contain. Other TR ligands such as those encompassed by Formula 1 described herein and known in the prior art, can also be used for thegeneration of co-crystals of TR LBD and TR ligands. Of the compounds encompassed by Formula 1 it is generally desirable to use at least one ligand that has at least one bromo- or iodo- substitution at the R.sub.3, R.sub.5, R.sub.3 ' or R.sub.5 'position, preferably such compounds will be have at least two such substitutions and more preferably at least 3 such substitutions. As described herein, such substitutions are advantageously used as heavy atoms to help solve the phase problem for thethree dimensional structure of the TR LBD and can be used as a generalized method of phasing using a halogen (e.g. I or Br) substituted ligand, especially for nuclear receptors.

Typically purified LBD, such as TR LBD, is equilibrated at a saturating concentration of ligand it a temperature that preserves the integrity of the protein. Ligand equilibration can be established between 2 and 37.degree. C., although thereceptor tends to be more stable in the 2-20.degree. C. range.

Preferably crystals are made with the hanging drop methods detailed herein. Regulated temperature control is desirable to improve crystal stability and quality. Temperatures between 4 and 25.degree. C. are generally used and it is oftenpreferable to test crystallization over a range of temperatures. In the case of TR it is preferable to use crystallization temperatures from 18 to 25.degree. C., more preferably 20 to 23.degree. C., and most preferably 22.degree. C.

Complexes of the TR-.alpha. LBD with a variety of agonists, including T.sub.3, IpBr.sub.2, Dimit, and Triac, are prepared with by methods described herein. For example, cocrystals of the rTR-.alpha. LBD, with ligand prebound, are prepared byvapor diffusion at ambient temperature from 15% 2-methyl-2,4-pentanediol (MPD). The crystals are radiation sensitive, and require freezing to measure complete diffraction data. On a rotating anode X-ray source, the crystals diffract to .about.3 .ANG.;synchrotron radiation extends the resolution limit significantly, to as high as 2.0 .ANG. for T.sub.3 cocrystals. The composition of the thyroid hormone, combined with the ability to prepare and cocrystallize the receptor complexed with a variety ofanalogs, permitted the unusual phasing strategy. This phasing strategy can be applied to the ligands of the nuclear receptors described therein by generating I and Br substitutions of such ligands. In this strategy, cocrystals of the TR LBD containingfour hormone analogs that differ at the 3,5, and 3' positions (T.sub.3, IpBr.sub.2, Dimit, and Triac) provided isomorphous derivatives. For this set of analogs, the halogen substituents (2Br and 3I atoms) function as heavy atoms, while the Dimitcocrystal (3 alkyl groups) acts as the parent. The initial 2.5 .ANG. multiple isomorphous replacement/anomalous scattering/density modified electron density map allowed the LBD to be traced from skeletons created in the molecular graphics program O5(Jones, T. A. et al., ACTA Cryst, 47:110-119 (1991), incorporated by reference herein). A model of the LBD was built in four fragments, Arg157-Gly184, Trp186-Gly197, Ser199-Pro205, and Val210-Phe405, and refined in XPLOR using positional refinement andsimulated annealing protocols. Missing residues were built with the aid of difference density. The final model was refined to R.sub.cryst =21.8% and R.sub.free =24.4% for data from 15.0 to 2.2 .ANG., see Table 6. The human TR-.beta. LBD model wasresolved by molecular replacement of the Tr-.alpha. LBD coordinates. The structure is based on E202 to D461 with a his-tag at the N-terminus. The final model was refined to R.sub.cryst =25.3% and R.sub.free =28.9% for data from 30.0 to 2.4 .ANG.+, seeTable 7.

This; phasing strategy can be applied to the ligands of the nuclear receptors described herein by generating I and Br substitutions of such ligands.

THREE DIMENSIONAL STRUCTURE OF TR LBD

Architecture of TR LBD

As an example of the three dimensional structure of a nuclear receptor, the folding of the TR-.alpha..sub.1 LBD is shown in FIG. 4. The TR-.alpha. LBD consists of a single structural domain packed in three layers, composed of twelve.alpha.-helices, H1-12, and four short .beta.-strands, S1-4, forming a mixed .beta.-sheet. The buried hormone and three antiparallel .alpha.-helices, H5-6, H9, and H10, form the central layer of the domain, as shown in FIG. 4. H1, H2, H3 and S1 formone face of the LBD, with the opposite face formed by H7, H8, H11, and H12. The first 35 amino acids of the N-terminus (Met122-Gln156) are not visible in the electron density maps. The three dimensional structure of the heterodimeric RXR:TR DNA-bindingdomains bound to DNA, amino acids Met 122-Gln151 of the TR DBD make extensive contacts with the minor groove of the DNA8. The five disordered amino acids (Arg152-Gln156), which reside between the last visible residue of the TR DBD and the first visibleresidue of ihe LBD likely represent the effective "hinge" linking the LBD and the DBD in the intact receptor.

The predominantly helical composition and the layered arrangement of secondary structure is identical to that of the unliganded hRXR.alpha., confirming the existence of a common nuclear receptor fold between two nuclear receptors.

The TR LBD is visible beginning at Arg157, and continues in an extended coil conformation to the start of H1. A turn of .alpha.-helix, H2, covers the hormone binding cavity, immediately followed by short .beta.-strand, S1, which forms the edgeof the mixed .beta.-sheet, parallel to S4, the outermost of the three antiparallel strands. The chain is mostly irregular until H3 begins, antiparallel to H1. H3 bends at Ile221 and IIe222, residues which contact the ligand. The chain turns almost90.degree. at the end of H3 to form an incomplete .alpha.-helix, H4. The first buried core helix, H5-6, follows, its axis altered by a kink near the ligand at Gly 253. The helix is composed of mostly hydrophobic sidechains interrupted by two strikingexceptions: Arg262 is solvent inaccessible and interacts with the ligand carboxylate (1-substituent), and Glu256 meets Arg329 from H9 and Arg375 from H11 in a polar invagination. H5-6 terminates in a short .beta.-strand, S2, of the four strand mixedsheet. S3 and S4 are joined through a left-handed turn, and further linked by a salt bridge between Lys284 and Asp272. Following S4, H7 and H8 form an L, stabilized by a salt bridge between Lys268 and Asp277. The turn between H7 and H8 adopts anunusual conformation, a result of interaction with ligand and its glycine rich sequence. H9 is the second core helix. antiparallel to the neighboring H5-6. Again, two buried polar sidechains are found, Glu315 and Gln320. Glu315 forms a buried saltbridge with His358 and Arg356. The oxygen of Gln320 forms a hydrogen bond with the buried sidechain of His 175. The chain then switches back again to form H10, also antiparallel to H9. H11 extends diagonally across the full length of the molecule. Immediately after H11, the chain forms a type II turn, at approximately 90.degree. to H11. The chain then turns again to form H 12, which packs loosely against H3 and H11 as part of the hormone or ligand binding cavity. The final five amino acids atthe C-terminus, Glu406-Val410, are disordered. The architecture of the TR-.beta., LBD is identical to that of the TR-.alpha. LBD, with two significant differences. An additional helix is present at the N-terminus (residues Glu202-Ile208), which ispart of the DBD, and packs antiparallel to H10. Following the helix is a two residue turn (Gly209-His210) continuing into an extended coil to he start of H1, as seen in the TR-.alpha. LBD. A further difference occurs in the irregular conformationadopted between H2 and H3. In the TR-.alpha. LBD, residue Gly197-Asp211 form a loop that packs against the receptor, contacting helices H7, H8, H11, and the loop between H11 and H12. In the TR-.beta. LBD, only the ends of the loop are ordered, withthe stretch Ala253-Lys263 disordered. In addition to these residues, the residues of the His-tag at the N-terminus, and the final residue at the C-terninus, Asp461, are disordered.

TR LBD's Ligand Binding Cavity as an Example of a Nuclear Receptor's Buried Ligand Cavity

The three dimensional structure of the TR LBD leads to the startling finding that ligand binding cavity of the LBD is solvent inaccessible when a T3 or its isostere is bound to the LBD. This surprising result leads to a new model of nuclearreceptor three dimensional structure and function, as further described herein, particularly in the sections elucidating the computational methods of ligand design and the application of such methods to designing nuclear receptor synthetic ligands thatcontain extended positions that prevent normal activation of the activation domain.

Dimit, the ligand bound to the receptor, is an isostere of T.sub.3 and a thyroid hormone agonist. Therefore the binding of Dimit should reflect that of T.sub.3, and the Dimit-bound receptor is expected to be the active conformation of TR. Theligand is buried within the receptor, providing the hydrophobic core for a subdomain of the protein, as shown in FIG. 5 a and b. H5-6 and H9 comprise the hydrophobic core for the rest of the receptor.

An extensive binding cavity is constructed from several structural elements. The cavity is enclosed from above by H5-6 (Met 256-Arg266), from below by H7 and H8 and the intervening loop (Leu287-Ile299), and along the sides by H2 (185-187), bythe turn between S3 and S4 (Leu276-Ser277), by H3 (Phe215-Arg228), by H11 (His381-Met388) and by H12 (Phe401-Phe405). The volume of the cavity defined by these elements, calculated by GRASP (Columbia University, USA) (600 .ANG.3), is essentially thevolume of the hormone (530 .ANG.). The change in volume can be exploited for ligand design as described herein. The remaining volume is occupied by water molecules surrounding the amino-propionic acid substituent. FIG. 6 depicts various contacts (orinteractions) between TR's LBD and the ligand.

The planes of the inner and outer (prime ring) rings of the ligand are rotated from planarity about 60.degree. with respect to each other, adopting the 3'-distal conformation (in which the 3' substituent of the outer ring projects down and awayfrom the inner ring). The amino-propionic acid and the outer phenolic ring assume the transoid conformation, each on opposite sides of the inner ring. The torsion angle .lambda..sub.1 for the amino-propionic acid is 300.degree..

The amino-propionic acid substituent is packed loosely in a polar pocket formed by side chains from H2, H4 and S3. The carboxylate group forms direct hydrogen bonds with the guanidium group of Arg228 and the amino N of Ser277. In addition,Arg262, Arg266 and Asn179 interact with the carboxylate through water-mediated hydrogen bonds. The three arginine residues create a significantly positive local electrostatic potential, which may stabilize the negative charge of the carboxylate. Nohydrogen bond is formed by the amino nitrogen. The interactions of the amino-propionic acid substituent are consistent with the fact that Triac, which lacks the amino nitrogen, has a binding affinity equal to that of T.sub.3, indicating that the aminonitrogen and longer aliphatic chain of T.sub.3 do not contribute greatly to binding affinity.

The biphenyl ether, in contrast, is found buried within the hydrophobic core. The inner ring packs in a hydrophobic pocket formed by H3, H5-6, and S3. Pockets for the 3- and 5-methyl substituents are not completely filled, as expected since thevan der waals radius of methyl substituent for Dimit is smaller than the iodine substituent provided by the thyroid hormone T.sub.3. Such pockets are typically 25 to 100 cubic angstroms (although smaller pocket for substitutes are contemplated in the 40to 80 cubic angstrom range) and could be filled more tightly with better fitting chemical substitutions, as described herein.

The outer ring packed tightly in a pocket formed by H3, H5-6, H7, H8, H11 and H12, and the loop between H7 and H8. The ether oxygen is found in a hydrophobic environment defined by Phe218, Leu287, Leu276, and Leu292. The absence of a hydrogenbond to the ether oxygen is consistent with its role in establishing the correct stereochemistry of the phenyl rings, as suggested by potent binding of hormone analogs with structurally similar linkages possessing reduced or negligible hydrogen bondingcapability. The 3'-isopropyl substituent contacts Gly290 and 291. The presence of glycine at this position in the pocket can explain the observed relationship between activity and the size of 3'-substituenls. Activity is highest for 3'-isopropyl, anddecreases with added bulk. The only hydrogen bond in the hydrophobic cavity is formed between the phenolic hydroxyl and His381 N.epsilon.2. The conformation of His381 is stabilized by packing contacts provided by Phe405, and Met256.

The presence of a 5' substituent larger than hydrogen affects the binding affinity for hormone. The more abundant thyroid hormone, 3,5,3',5'-tetraiodo-L-thyronine (T.sub.4), contains an iodine at this position, and binds the receptor with 2% ofthe affinity of T.sub.3. The structure suggests that discrimination against T.sub.4 is accomplished through the combination of steric conflict by Met256 and possibly the constraints imposed by the geometry of the hydrogen bond from His381 to thephenolic hydroxyl. The 5' position is a preferred location for introducing a chemical modification of C-H at the 5' of T3 or and TR agonist, as described herein, that produces an extension from the prime ring and results in the creation of an antagonistor partial agonist.

Deletion and antibody competition studies suggest the involvement of residues Pro162 to Val202 in ligand binding. The region does not directly contact hormone in the bound structure, although H2 packs against residues forming the polar pocketthat interacts with the amino-propionic acid group. One role for H2, then, is to stabilize these residues in the bound state, H2, with .beta.-strands S3 and S4, might also represent a prevalent entry point for ligand, since the amino-propionic acid ofthe ligand is oriented toward this region. Studies of receptor binding to T.sub.3 affinity matrices demonstrate that only a linkage to the amino-propionic acid is tolerated, suggesting that steric hindrance present in other linkages prevent binding. Furthermore, the crystallographic temperature factors suggest the coil and .beta.-strand region is most flexible part of the domain FIG. 7. Participation of this region, part of the hinge domain between the DBD and LBD, in binding hormone may providestructural means for ligand binding to influence DNA binding, since parts of the Hinge domain contact DNA.

TR LBD Transcriptional Activation Helix as an Example of a Nuclear Receptor Activation Domain

In addition to the startling finding that the ligand binding cavity is solvent inaccessible when loaded with a ligand, the activation helix of TR LBD presents a surface to the ligand cavity for interaction between at least one amino acid and thebound ligand. The C-terminal 17 amino acids of the TR, referred to as the activation helix or AF-2 (an example of an LBD activation domain), are implicated in mediating hormone-dependent transcriptional activation. Although, mutations of key residueswithin the domain decrease ligand-dependent activation it was unclear until the present invention whether such mutations directly affected ligand coordination. Although some mutations of this domain have been noted to reduce or abolish ligand binding,other mutations in more distant sites of the LBD have a similar effect.

Activation domains among nuclear receptors display an analogous three dimensional relationship to the binding cavity, which is a region of the LBD that binds the molecular recognition domain of a ligand, i.e. the activation domain presents aportion of itself to the binding cavity (but necessarily the molecular recognition domain of the ligand). Many nuclear receptors are expected to have such domains, including the retinoid receptors, RAR and RXR, the glucocorticoid receptor GR, and theestrogen receptor ER. Based upon the TR's sequence, the domain is proposed to adopt an amphipathic helical structure. .beta.-sheet or mixed secondary structures, could be present as activation domains in less related nuclear receptors.

Within the activation domain, the highly conserved motif .PHI..PHI.XE.PHI..PHI., where .PHI. represents a hydrophobic residue, is proposed to mediate interactions between the receptors and transcriptional coactivators. Several proteins havebeen identified which bind the TR in a hormone-dependent fashion. One of these, Trip1, is related to a putative yeast coactivator Sug1, and also interacts with both the C-terminal activation domain and a subset of the basal transcriptional machinery,suggesting a role in transactivation by the TR. Other proteins, such as RIP140, SRC1, (Onate, S. A. et. al., Science 270:1354-1357 (1995)) and TF-1 (see also Ledouarim, B., et. al., EMBO J. 14:2020-2033 (1995)), and GRIP-1 (Heery, E., et al., Nature387:733-736 (1997)) also interact with other nuclear receptors in a ligand dependent manner through the C-terminal domain. Binding of these proteins can be modulated using the TR ligands described herein especially those TR ligands with extensions thatsterically hinder the interaction between the highly conserved motif and other proteins.

The C-terminal activation domain of the TR forms an amphipathic helix, H12, which nestles loosely against the receptor to form part of the hormone binding cavity. The helix packs with the hydrophobic residues facing inward towards the hormonebinding cavity, and the charged residues, including the highly-conserved glutamate, extending into the solvent, as shown in FIG. 8. The activation helix of TR LBD presents Phe 401 to the ligand binding cavity and permits direct coordination with thehormone i.e. such amino acids interact with the ligand forming a van der waals contact with the plane of the outer phenyl ring. Phe 405 also interacts with His 381, perhaps stabilizing its hydrogen bonding conformation, i.e. a favorable hydrogen bondinteraction. Participation of Phe 401 and Phe 405 in binding hormone (explains how mutation of these residues decreases hormone binding affinity. Furthermore, the impact of these mutations on activation likely derives from a role in stabilizing thedomain in the bound structure through increased hydrogen bond interaction of dipole interactions. Glu 403 extends into the solvent, emphasizing its critical role in transactivation. In its observed conformation, presented on the surface as an orderedresidue, against a background of predominantly hydrophobic surface, Glu 403 is available to interact with activator proteins described herein, as shown in FIG. 9. The other charged residues, Glu 405 and Asp 406 are disordered, as the helix frays at Phe405.

Two other sequences in the TR, .tau.2 and .tau.3, activate transcription when expressed as fusion proteins with a DNA-binding domain. The sequences, discovered in the TRB, correspond to TR-.alpha. residues Pro158-Ile168 in H1 (.tau.2), andGly290-Leu3 19 in H8 and H9 (.tau.3). Unlike the C-terminal activation domain, .tau.2 and .tau.3 do not appear to represent modular structural units in the rat TR-.alpha. LBD, nor present a surface for protein-protein interactions: the criticalaspartate/glutamate residues of .tau.3 are located on two separate helices, and do not form a single surface; the charged residues of .tau.2 are engaged in ion pair interactions with residues of the LBD. Thus, .tau.2 and .tau.3 may not function asactivation domains in the context of the entire receptor.

Computational Methods for Designing a Nuclear Receptor LBD LIGAND

The elucidation of the three dimensional structure of a nuclear receptor ligand binding domain provides an important and useful approach for designing ligands to nuclear receptors using the computational methods described herein. By inspectingthe FIGURES it can be determined that the nuclear receptor ligand is bound in a water inaccessible binding cavity in the LBD and that chemical moieties can be added to selected positions on the ligand. Such chemical modifications, usually extensions,can fill up the binding cavity represented in the FIGURES for a tighter fit (or less water) or can be used to disrupt or make contacts with amino acids not in contact with the ligand before the chemical modification was introduced or represented in afigure of the three dimensional model of the LBD. Ligands that interact with nuclear superfamily members can act as agonists, antagonists and partial agonists based on what ligand-induced conformational changes take place.

Agonists induce changes in receptors that place them in an active conformation that allows them to influence transcription, either positively or negatively. There may be several different ligand-induced changes in the receptor's conformation.

Antagonists, bind to receptors, but fail to induce conformational changes that alter the receptor's transcriptional regulatory properties or physiologically relevant conformations. Binding of an antagonist can also block the binding andtherefore the actions of an agonist.

Partial agonists bind to receptors and induce only part of the changes in the receptors that are induced by agonists. The differences can be qualitative or quantitative. Thus, a partial agonist may induce some of the conformation changesinduced by agonists, but not others, or it may only induce certain changes to a limited extent.

Ligand-induced Conformational Changes

As described herein, the unliganded receptor is in a configuration that is either inactive, has some activity or has repressor activity. Binding of agonist ligands induces conformational changes in the receptor such that the receptor becomesmore active, either to stimulate or repress the expression of genes. The receptors may also have non-genomic actions. Some of the known types of changes and/or the sequelae of these are listed herein.

Heat Shock Protein Binding

For many of the nuclear receptors ligand binding induces a dissociation of heat shock proteins such that the receptors can form dimers in most cases, after which the receptors bind to DNA and regulate transcription.

Nuclear receptors usually have heat shock protein binding domains that present a region for binding to the LBD and can be modulated by the binding of a ligand to the LBD. Consequently, an extended chemical moiety (or more) from the ligand thatstabilizes the binding or contact of the heat shock protein binding domain with the LBD can be designed using the computational methods described herein to produce a partial agonist or antagonist. Typically such extended chemical moieties will extendpast and away from the molecular recognition domain on the ligand and usually past the buried binding cavity of the ligand.

Dimerization and Heterodimerization

With the receptors that are associated with the hsp in the absence of the ligand, dissociation of the hsp results in dimerization of the receptors. Dimerization is due to receptor domains in both the DBD and the LBD. Although the main stimulusfor dimerization is dissociation of the hsp, the ligand-induced conformational changes in the receptors may have an additional facilitative influence. With the receptors that are not associated with hsp in the absence of the ligand, particularly withthe TR, ligand binding can affect the pattern of dimerization/heterodimerization. The influence depends on the DNA binding site context, and may also depend on the promoter context with respect to other proteins that may interact with the receptors. Acommon pattern is to discourage monomer formation, with a resulting preference for heterodimer formation over dimer formation on DNA.

Nuclear receptor LBDs usually have dimerization domains that present a region for binding to another nuclear receptor and can be modulated by the binding of a ligand to the LBD. Consequently, an extended chemical moiety (or more) from the ligandthat disrupts the binding or contact of the dimerization domain can be designed using the computational methods described herein to produce a partial agonist or antagonist. Typically such extended chemical moieties will extend past and away from themolecular recognition domain on the ligand and usually past the buried binding cavity of the ligand.

DNA Binding

In nuclear receptors that bind to hsp, the ligand-induced dissociation of hsp with consequent dimer formation allows, and therefore, promotes DNA binding. With receptors that are not associated (as in the absence of ligand), ligand binding tendsto stimulate DNA binding of heterodimers and dimers, and to discourage monomer binding to DNA. However, ligand binding to TR, for example, tends to decrease dimer binding on certain DNA elements and has minimal to no effect on increasing heterodimerbinding. With DNA containing only a single half site, the ligand tends to stimulate the receptor's binding to DNA. The effects are modest and depend on the nature of the DNA site and probably on the presence of other proteins that may interact with thereceptors. Nuclear receptors usually have DBDs that present a region for binding to DNA and this binding can be modulated by the binding of a ligand to the LBD. Consequently, an extended chemical moiety (or more) from the ligand that disrupts thebinding or contact of the DBD can be designed using the computational methods described herein to produce a partial agonist or antagonist. Typically such extended chemical moieties will extend past and away from the molecular recognition domain on theligand and usually past the buried binding cavity of the ligand.

Repressor Binding

Receptors that are not associated with hsp in the absence of ligand frequently act as transcriptional repressors in the absence of the ligand. This appears to be due, in part, to transcriptional repressor proteins that bind to the LBD of thereceptors. Agonist binding induces a dissociation of these proteins from the receptors. This relieves the inhibition of transcription and allows the transcriptional transactivation functions of the receptors to become manifest.

Transcriptional Transactivation Functions

Ligand binding induces transcriptional activation functions in two basic ways. The first is through dissociation of the hsp from receptors. This dissociation, with consequent dimerization of the receptors and their binding to DNA or otherproteins in the nuclear chromatin allows transcriptional regulatory properties of the receptors to be manifest. This may be especially true of such functions on the amino terminus of the receptors.

The second way is to alter the receptor to interact with other proteins involved in transcription. These could be proteins that interact directly or indirectly with elements of the proximal promoter or proteins of the proximal promoter. Alternatively, the interactions could be through other transcription factors that themselves interact directly or indirectly with proteins of the proximal promoter. Several different proteins have been described that bind to the receptors in aligand-dependent manner. In addition, it is possible that in some cases, the ligand-induced conformational changes do not affect the binding of other proteins to the receptor, but do affect their abilities to regulate transcription.

Nuclear receptors or nuclear receptor LBDs usually have activation domains modulated in part by a co-activator/co-repressor system that coordinately functions to present a region for binding to DNA, and can be modulated by the binding of a ligandto the LBD. Consequently, an extended chemical moiety (or more) from the ligand that disrupts the binding or contact of the activation domain with co-activator and/or co-repressor can be designed using the computational methods described herein toproduce a partial agonist or antagonist. For instance, an agonist can be designed and/or selected which (1) blocks binding and/or dissociates co-repressor, and/or (2) promotes binding and/or association of a co-activator. An antagonist can be designedwhich (1) promotes binding and/or association of co-repressor, and/or (2) promotes binding and/or association of co-activator. Ratios of agonists and antagonists may be used to modulate transcription of the gene of interest. Selection can beaccomplished in binding assays that screen for ligands having the desired agonist or antagonist properties, including such ligands which induce conformational changes as described below. Suitable assays for such screening are described herein and inShibata, H., et al. (Recent Prog. Horm. Res. 52:141-164 (1997)); Tagami, T., et al. (Mol. Cell Biol. 17(5):2642-2648 (1997)); Zhu, X G., et al. (J. Biol. Chem. 272(14):9048-9054 (1997)); Lin, B. C., et al. (Mol. Cell Biol. 17(10):6131-6138 (1997));Kakizawa, T., et al. (J. Biol. Chem. 272(38):23799-23804 (1997)); and Chang, K. H., et al. (Proc. Natl. Acad. Sci. USA 94(17):9040-9045 (1997)). Typically such extended chemical moieties will extend past and away from the molecular recognitiondomain on the ligand and usually past the buried binding cavity of the ligand and in the direction of the activation domain, which is often a helix as seen in the three dimensional model shown in the FIGURES in two dimensions on paper or moreconveniently on a computer screen.

Ligand-Induced Conformational Change

Plasma proteins bind hormones without undergoing a conformational change through a static binding pocket formed between monomers or domains. For example, the tetrameric thyroid-binding plasma protein transthyretin forms a solvent-accessiblehormone-binding channel at the oligomer interface. The structure of the protein is unchanged upon binding hormone with respect to the appearance of a buried binding cavity with a ligand bound.

However, the structural role for a ligand bound to a nuclear receptor LBD, like rat TR-.alpha. LBD, predicts that the receptor would differ in the bound and unbound states. In the absence of hormone, the receptor would possess a cavity at itscore, uncharacteristic of a globular protein. A ligand (e.g. hormone) completes the hydrophobic core of the active receptor after it binds to the nuclear receptor. Ligand binding by the receptor is a dynamic process, which regulates receptor functionby inducing an altered conformation.

An exact description of the hormone-induced conformational changes requires comparison of the structures of the liganded and the unliganded TR. The structure of the unliganded human RXR.alpha. may substitute as a model for the unliganded TR. The rat TR-.alpha. LBD and human RXR.alpha. LBDs adopt a similar fold, and it is likely that the structural similarity extends to the conformational changes after ligand binding.

There are three major differences between the two structures, which indeed appear to be the result of ligand binding. First, the bound rat TR-.alpha. LBD structure is more compact, with the hormone tightly packed within the hydrophobic core ofthe receptor. By contrast, the unliganded human RXR.alpha. LBD contains several internal hydrophobic cavities. The presence of such cavities is unusual in folded proteins, and is likely a reflection of the unliganded state of the receptor. Two ofthese cavities were proposed as possible binding sites for 9-cis retinoic acid, though these multiple sites only partly overlap with the single buried binding cavity observed in the liganded rat TR-.alpha. LBD.

The second difference involves H11 in the rat TR-.alpha. LBD, which contributes part of the hormone binding cavity. H11, continuous in the rat TR-.alpha. LBD, is broken at Cys 432 in the RXR, forming a loop between H10 and H11 in thehRXR.alpha.. This residue corresponds to His381 in the TR, which provides a hydrogen bond to the outer ring hydroxyl of the ligand. Furthermore, the hormone binding cavity occupied by ligand in the rat TR-.alpha. LBD is interrupted in the hRXR.alpha. by the same loop, forming an isolated hydrophobic pocket in the RXR with H6 and H7. In the bound rat TR-.alpha. LBD, the corresponding helices H7 and H8 are contiguous with the binding pocket, and enclose the hormone binding cavity from below.

The third difference between the two receptors is the position of the C-terminal activation domain. While the C-terminal activation domain forms .alpha.-helices in both receptors, the domain in the rat TR-.alpha. LBD follows a proline-richturn, and lies against the receptor to contribute part of the binding cavity. In contrast, the activation domain in the unliganded hRXR.alpha., is part of a longer helix which projects into the solvent.

These differences lead to a model for an alternate conformation of the TR LBD assumed in the absence of ligand. In the unliganded TR, the subdomain of the receptor surrounding the hormone binding cavity is loosely packed, with the binding cavityoccluded by a partly unstructured H11 providing a partial core for the receptor.

Upon binding hormone, residues which form a coil in the unbound receptor engage the ligand, and continues H11. The ordering of H11 could unblock the hydrophobic cavity, allowing H7 and H8 to interact with hormone. The extended hydrophobiccavity then collapses around the hormone, generating the compact bound structure.

It is possible to predict ligand-induced conformational changes in the C-terminal activation domain that rely, in part, on an extended structure in the unliganded TR that repacks upon ligand binding. The ligand-induced conformation change can besubtle since the amino acid sequence of the rat TR-.alpha. in the turn (393-PTELFPP-399) significantly reduces the propensity of the peptide chain of the rat TR-.alpha. to form an .alpha.-helix and therefore repacking can be accomplished with a minorchange in volume.

After the ligand-induced conformational change occurs, it is likely that the conformation of the C-terminal activation domain in the bound structure changes packing compared to the unbound form of the receptor. Binding of the ligand improves thestability of the activation domain. The activation domain packs loosely even in the bound structure, as measured by the distribution of packing interactions for the entire LBD. The packing density for the activation domain, defined as the number ofatoms within 4.5 .ANG., is 1.5 standard deviations below the mean. For comparison, another surface helix, H1, is 0.5 standard deviations below the mean and the most poorly packed part of the structure, the irregular coil from residues Ile196-Asp206, is2.0 standard deviations below the mean. Moreover, the majority of packing contacts for the C-terminal domain in the bound receptor are provided either by residues which interact with ligand, such as His381, or by the ligand itself. The conformation ofthese residues can be expected to be different in the bound and unbound receptors, and by extension the conformation of C-terminal activation domain which relies upon these interactions. Without the stabilization provided by a bound ligand, it is likelythat the C-terminal activation domain is disordered prior to hormone binding.

The interrelation of ligand-induced conformational changes is evident as described herein. For example, His381 from H11 and Phe405 from H12 interact in the bound structure to provide a specific hydrogen bond to the phenolic hydroxyl. Theligand-induced changes which affect H11 and H12 are reinforcing, and lead to the formation of the compact, bound state.

Comparison of the TR-.alpha. and TR-.beta. LBD structures shows similar packing of the helices when complexed with the ligand Triac.

COMPUTATIONAL METHODS USING THREE DIMENSIONAL MODELS AND EXTENSIONS OF LIGANDS

The three-dimensional structure of the liganded TR receptor is unprecedented, and will greatly aid in the development of new nuclear receptor synthetic ligands, such as thyroid receptor antagonists and improved agonists, especially those thatbind selectively to one of the two TR isoforms (.alpha. or .beta.). In addition, this receptor superfamily is overall well suited to modern methods including three-dimensional structure elucidation and combinatorial chemistry such as those disclosed inEP 335 628, U.S. Pat. No. 5,463,564, which are incorporated herein by reference. Structure determination using X-ray crystallography is possible because of the solubility properties of the receptors. Computer programs that use crystallography datawhen practicing the present invention will enable the rational design of ligand to these receptors. Programs such as RASMOL can be used with the atomic coordinates from crystals generated by practicing the invention or used to practice the invention bygenerating three dimensional models and/or determining the structures involved in ligand binding. Computer programs such as INSIGHT and GRASP allow for further manipulation and the ability to introduce new structures. In addition, high throughputbinding and bioactivity assays can be devised using purified recombinant protein and modern reporter gene transcription assays described herein and known in the art in order to refine the activity of a CDL.

Generally the computational method of designing a nuclear receptor synthetic ligand comprises two steps:

1) determining which amino acid or amino acids of a nuclear receptor LBD interacts with a first chemical moiety (at least one) of the ligand using a three dimensional model of a crystallized protein comprising a nuclear receptor LBD with a boundligand, and

2) selecting a chemical modification (at least one) of the first chemical moiety to produce a second chemical moiety with a structure to either decrease or increase an interaction between the interacting amino acid and the second chemical moietycompared to the interaction between the interacting amino acid and the first chemical moiety.

As shown herein, interacting amino acids form contacts with the ligand and the center of the atoms of the interacting amino acids are usually 2 to 4 angstroms away from the center of the atoms of the ligand. Generally these distances aredetermined by computer as discussed herein and in McRee 1993, however distances can be determined manually once the three dimensional model is made. Examples of interacting amino acids are described in Appendix 2. See also Wagner et al., Nature378(6558):670-697 (1995) for stereochemical figures of three dimensional models. More commonly, the atoms of the ligand and the atoms of interacting amino acids are 3 to 4 angstroms apart. The invention can be practiced by repeating steps 1 and 2 torefine the fit of the ligand to the LBD and to determine a better ligand, such as an agonist. As shown in the FIGURES the three dimensional model of TR can be represented in two dimensions to determine which amino acids contact the ligand and to selecta position on the ligand for chemical modification and changing the interaction with a particular amino acid compared to that before chemical modification. Structural comparison of LBD isoforms complexed with the same or similar ligand permitidentification of fiducial and adjustable amino acids that can be exploited in designing isoform-specific ligands through chemical modification. "Fiducial" refers to amino acids that form rigid features of the ligand binding cavity. "Adjustable" refersto amino acids that form less rigid features of the ligand binding cavity. The chemical modification may be made using a computer, manually using a two dimensional representation of the three dimensional model or by chemically synthesizing the ligand. The three dimensional model may be made using Appendix 2 and the FIGURES. As an additional step, the three dimensional model may be made using atomic coordinates of nuclear receptor LBDs from crystallized protein as known in the art, see McRee 1993referenced herein.

The ligand can also interact with distant amino acids after chemical modification of the ligand to create a new ligand. Distant amino acids are generally not in contact with the ligand before chemical modification. A chemical modification canchange the structure of the ligand to make as new ligand that interacts with a distant amino acid usually at least 4.5 angstroms away from the ligand. Often distant amino acids will not line the surface of the binding cavity for the ligand, as they aretoo far away from the ligand to be part of a pocket or surface of the binding cavity.

The interaction between an atom of a LBD amino acid and an atom of an LBD ligand can be made by any force or attraction described in nature. Usually the interaction between the atom of the amino acid and the ligand will be the result of ahydrogen bonding interaction, charge interaction, hydrophobic interaction, van der waals interaction or dipole interaction. In the case of the hydrophobic interaction it is recognized that this is not a per se interaction between the amino acid andligand, but rather the usual result, in part, of the repulsion of water or other hydrophilic group from a hydrophobic surface. Reduction or enhancement of the interaction of the LBD and a ligand can be measured by standard binding procedures,calculating or testing binding energies, computationally or using thermodynamic or kinetic methods as known in the art.

Chemical modifications will often enhance or reduce interactions of an atom of a LBD amino acid and an atom of an LBD ligand. Steric hinderance will be a common means of changing the interaction of the LBD binding cavity with the activationdomain. Chemical modifications are preferably introduced at C--H, C-- and C--OH position in ligands, where the carbon is part of the ligand structure which remains the same after modification is complete. In the case of C--H, C-- could have 1, 2 or 3hydrogens, but usually only one hydrogen will be replaced. The H or OH are removed after modification is complete and replaced with the desired chemical moiety.

Because the thyroid receptor is a member of the larger superfamily of hormone-binding nuclear receptors, the rules for agonist and antagonist development will be recognized by one skilled in the art as useful in designing ligands to the entiresuperfamily. Examining the structures of known agonists and antagonists of the estrogen and androgen receptors supports the generality of antagonist mechanism of action as shown in FIG. 10.

The overall folding of the receptor based on a comparison of the reported structure of the unliganded RXR and with amino acid sequences of other superfamily members reveals that the overall folding of receptors of the superfamily is similar. Thus, it is predicted from the structure that there is a general pattern of folding of the nuclear receptor around the agonist or antagonist ligand.

The three dimensional structure of a nuclear receptor with a ligand bound leads to the nonobvious observation that a nuclear receptor folds around agonist ligands, as the binding cavity fits the agonist, especially the agonist's molecularrecognition domain, and antagonists commonly have chemical structures that extend beyond the ligand, especially the agonist, and would prohibit folding of the receptor around the ligand to form a buried binding cavity or other groups that have the sameeffect. The location of the extension could affect the folding in various ways as indicated by the structure. Such extensions on antagonists are shown in FIG. 10 for various receptors and compared to the corresponding agonist.

For example, an extension towards the carboxy-terminal activation helix affects the packing/folding of this helix into the body of the receptor. This in turn can affect the ability of this portion of the nuclear receptor to interact with otherproteins or other portions of the receptor, including transcriptional transactivation functions on the opposite end of the linear receptor, or the receptor's amino terminus that may interact directly or indirectly with the carboxy-terminaltransactivation domain (including helix 12). Extensions in this direction can also affect the packing of helix 11 of TR (or its analogous helix in nuclear receptors) into the body of the receptor and selectively affect dimerization andheterodimerization of receptors. An extension pointing towards helix 1 can affect the relationship of the DNA binding domain and hinge regions of the receptors with the ligand binding domain and selectively or in addition affect the receptors' bindingto DNA and/or interactions of receptors with proteins that interact with this region of the receptor. Other extensions towards helix 11 can be made to affect the packing of this helix and helices 1 and 10 and thereby homo- and hetero-dimerization. Suchchemical modifications can be assessed using the computational methods described herein. It is also possible that, in some cases, extensions may protrude through the receptor that is otherwise completely or incompletely folded around the ligand. Suchprotruding extensions could present a steric blockade to interactions with co-activators or other proteins.

The three dimensional structure with the ligand buried in the binding cavity immediately offers a simple description of a nuclear receptor that has a binding cavity that contains hinges and a lid, composed of one or more structural elements, thatmove to accommodate and surround the ligand. The ligand to TR can be modified on specific sites with specific classes of chemical groups that will serve to leave the lid and hinge region in open, partially open or closed states to achieve partialagonist or antagonist functions. In these states, the biological response of the TR is different and so the structure can be used to design particular compounds with desired effects.

Knowledge of the three-dimensional structure of the TR-T.sub.3 complex leads to a general model for agonist and antagonist design. An important novel feature of the structural data is the fact that the T.sub.3 ligand is completely buried withinthe central hydrophobic core of the protein. Other ligand-receptor complexes belonging to the nuclear receptor superfamily will have a similarly buried ligand binding site and therefore this model will be useful for agonist/antagonist design for theentire superfamily.

When design of an antagonist is desired, one needs either to preserve the important binding contacts of natural hormone agonist while incorporating an "extension group" that interferes with the normal operation of the ligand-receptor complex orto generate the requisite binding affinity through the interactions of the extensions with receptor domains.

The model applied to antagonist design and described herein is called the "Extension Model." Antagonist compounds for nuclear receptors should contain the same or similar groups that facilitate high-affinity binding to the receptor, and inaddition, such compounds should contain a side chain which may be large and/or polar. This side chain could be an actual extension, giving it bulk, or it could be a side group with a charge function that differs from the agonist ligand. For example,substitution of a CH.sub.3 for CH.sub.2 OH at the 21-position, and alteration at the 11-position from an OH group to a keto group of cortisol generates glucocorticoid antagonist activity (Robsseau, G. G., et. al., J. Mol. Biol. 67:99-115 (1972)). However, in most cases effective antagonists have more bulky extensions. Thus, the antiglucocorticoid (and antiprogestin) RU486 contains a bulky side group at the 11-position (Horwitz, K. B. Endocrine Rev. 13:146-163 (1992)). The antagonist compoundwill then bind within the buried ligand binding site of the receptor with reasonably high affinity (100 nM), but the extension function will prevent the receptor-ligand complex from adopting the necessary conformation needed for transcription factorfunction. The antagonism (which could be in an agonist or antagonist) may manifest itself at the molecular level in a number of ways, including by preventing receptor homo/heterodimer formation at the HRE, by preventing coactivator binding to receptormonomers, homodimers or homo/heterodimers, or by a combination of these effects which otherwise prevent transcription of hormone responsive genes mediated by ligand-induced effects on the HRE. There are several antagonist compounds for nuclear receptorsin the prior art (see also Horwitz, K. B., Endocrine Rev. 13:146-163 (1992), Raunnaud J. P. et. al., J. Steroid Biochem. 25:811-833 (1986), Keiel S., et. al., Mol. Cell. Biol. 14:287-298 (1994) whose antagonist function can be explained by theextension hypothesis. These compounds are shown in FIG. 10 along with their agonist counterparts. Each of these antagonists contains a large extension group attached to an agonist or agonist analogue core structure. Importantly, these antagonistcompounds, were discovered by chance and not designed with a structure-function hypothesis such as the extension principle.

One method of design of a thyroid antagonist using the extension hypothesis is provided below as a teaching example. The three-dimensional structure of the TR-.alpha. Dimit complex combined with structure-activity data published in the priorart, especially those reference herein, can be used to establish the following ligand-receptor interactions which are most critical for high-affinity ligand binding. A physical picture of these interactions is shown in FIG. 6. The figure describes theisolated essential contacts for ligand binding. Because the ligand is buried in the center of the receptor, the structural spacing between these isolated interactions is also important. Thus, our present knowledge of this system dictates that, for thisexample, a newly designed ligand for the receptor must contain a thyronine structural skeleton, or two substituted aryl groups joined by a one-atom spacer.

The general structure for an antagonist designed by the extension hypothesis is exemplified in the following general description of the substituents of a TR antagonist (referring to Formula 1): R1 can have anionic groups such as a carboxylate,phosphonate, phosphate, sulfate or sulfite and is connected to the ring with a 0 to 3 atom linker, comprising) one or more C, O, N, S atoms, and preferably a 2 carbon linker. Such R1 can be optionally substituted with an amine (e.g. --NH2). R3 and R5are small hydrophobic groups such as --Br, --I, or --CH3. R3 and R5 can be the same substituents or different. R.sub.3 ' can be a hydrophobic group that may be larger than those of R3 and R5, such as --I, --CH3, -isopropyl, -phenyl, -benzyl, 5 and 6ring heterocycles. R.sub.4 ' is a group that can participate in a hydrogen bond as either a donor or acceptor. Such groups include --OH, --NH.sub.2, and --SH. R.sub.5 ' is an important extension group that makes this compound an antagonist. R.sub.5 'can be a long chain alkyl (e.g. 1 to 9 carbons, straight chain or branched), aryl (benzyl, phenyl and substituted benzyl and phenyl rings (e.g with halogen, alkyl (1 and 5 carbons) and optionally connected to the ring by a --CH2--), heterocycle (e.g. 5or 6 atoms, preferably 5 carbons and 1 nitrogen, or five carbons), which can optionally include polar (e.g. --OH, --NH.sub.2, and --SH), cationic (e.g. --NH3, N(CH)3), or anionic (carboxylate, phosphonate, phosphate or sulfate) groups. R.sub.5 ' canalso be a polar (e.g. --OH, --NH.sub.2, and --SH), cationic (e.g. --NH3, --N(CH3)3), and anionic (carboxylate, phosphonate, phosphate or sulfate) groups. X is the spacer group that appropriately positions the two aromatic rings. This group is usually aone-atom spacer, such as O, S, SO, SO2, NH, NZ where Z is an alkyl, CH2, CHOH, CO, C(CH3)OH, and C(CH3)(CH3). X also may be NR.sub.7, CHR.sub.7, CR.sub.7, R.sub.7, where R.sub.7, is an alkyl, aryl or 5- or 6-membered heterocyclic aromatic. R2, R6, R2'and R6' can be --F, and --Cl and are preferably H.

A TR ligand can also be described as a substituted phenylated 3,5 diiodo tyrosine with substituted R5' and R3' groups. R5' can be a long chain alkyl (e.g. 4 to 9 carbons, straight chain or branched), aryl (benzyl, phenyl and substituted benzyland phenyl rings (e.g with halogen, alkyl (1 and 5 carbons) and optionally connected to the ring by a --CH2--), heterocycle (e.g. 5 or 6 atoms, preferably 5 carbons and 1 nitrogen, or five carbons), which can optionally include polar (e.g. --OH,--NH.sub.2, and --SH), cationic (e.g. --NH3, N(CH)3), or anionic (carboxylate, phosphonate, phosphate or sulfate) groups. R5' can also be a polar (e.g. --OH, --NH.sub.2, and --SH), cationic (e.g. --NH3, N(CH)3), and anionic (carboxylate, phosphonate,phosphate or sulfate) groups. R3' can be -IsoPr, halogen, --CH3, alkyl (1 to 6 carbons) or aryl (benzyl, phenyl and substituted benzyl and phenyl rings (e.g with halogen, alkyl (1 and 5 carbons) and optionally connected to the ring by a --CH2--),heterocycle (e.g. 5 or 6 atoms, preferably 5 carbons and 1 nitrogen, or five carbons), which can optionally include polar (e.g. --OH, --NH.sub.2, and --SH), cationic (e.g. --NH3, N(CH)3), or anionic (carboxylate, phosphonate, phosphate or sulfate)groups.

A TR antagonist can also be a modified T.sub.3 agonist (having a biphenyl structure) wherein R.sub.5 ' is alkyl, aryl, 5- or 6-membered heterocyclic aromatic, heteroalkyl, heteroaryl, arylalkyl, heteroaryl alkyl, polyaromatic, polyheteroaromatic,polar or charged groups, wherein said R.sub.5 ' may be substituted with polar or charged groups. The R5' groups are defined, as described herein.

Using these methods the ligands of this example preferably have the following properties:

1. The compounds should bind to the TR with high affinity (for example 100 nM).

2. The compounds should bind the receptor in the same basic orientation as the natural hormone.

3. The extension group R5' should project toward the activation helix (C-terminal helix) of the receptor.

4. The appropriate substituent at R5' should perturb the activation helix from its optimal local structure needed for mediating transcription.

Antagonists may also be designed with multiple extensions in order to block more than one aspect of the folding at any time.

TR ligands (e.g. super agonists) can be designed (and synthesized) to enhance the interaction of at least one amino acid with at least one chemical moiety on the ligand's molecular recognition domain. One method is to enhance the charge andpolar interactions by replacing the carboxylate of T.sub.3 (R1 position) with phosphonate, phosphate, sulfate or sulfite. This enhances the interaction with Arg 262, Arg 266 and Arg 228. The interaction of at least one amino acid with at least onechemical moiety on the ligand's molecular recognition domain can also be enhanced by increasing the size of R1 group to fill the space occupied by water when Dimit is bound (referring to R1). Preferably the group has a complementary charge andhydrophobicity to the binding cavity.

Another way of improving the interaction of at least one amino acid with at least one chemical moiety on the ligand's molecular recognition domain is to restrict the conformation of the dihedral angle between the two phenyl rings of the thyronineligand in solution. In solution the planes of two phenyl rings are orthogonal where the dihedral angle is 90.degree.. In the TR Dimit structure, the dihedral angle is close to 60.degree.. A TR ligand design that fixes the angle between the two phenylrings will lead to tighter binding. Such a ligand may be made by connecting the R6' and the R5 positions of a thyronine or a substituted thyronine-like biphenyl. The size of the cyclic connection can fix the angle between the two phenyl rings. Referring specifically to Formula 1, the following cyclic modifications are preferred: 1) R.sub.5 is connected to R.sub.6 ', 2) R.sub.3 is connected to R.sub.2 ' or 3) R.sub.5 is connected to R.sub.6 ' and R.sub.3 is connected to R2'. The connectionscan be made by an alkyl or heteroalkyl chain having between 1 to 6 atoms and preferably from 2 to 4 carbon atoms or other atoms. Any position of the heteroalkyl chain can be N, O, P or S. The S and P heteroatoms along said heteroalkyl chain are in anyof their possible oxidative states. The N heteroatom or any carbon along the alkyl or heteroalkyl chain may have one or more Z substituents, wherein Z is alkyl, heteroalkyl, aryl, heteroaryl, 5- or 6-membered heterocyclic aromatic. These compounds canbe claimed with the proviso that Formula 1 does not include any prior art compound as of the priority filing date of this application.

The interaction of at least one amino acid with at least one chemical moiety on the ligand's molecular recognition domain can also be enhanced by selecting a chemical modification that fills the unfilled space between a TR ligand and the LBD inthe area of the bridging oxygen (such as in T3, Triac or Dimit). Thus, a slighter larger moiety that replaces the ether oxygen can enhance binding. Such a linker may be a mono- or geminal-disubstituted carbon group. A group approximately the same sizeas oxygen but with greater hydrophobicity is preferred as well as small, hydrophobic groups for the disubstituted carbon.

Compounds of Formula I or derivatives thereof that modulate TR activity also may be designed and selected to interact with a conformationally constrained structural feature of a TR LBD that is conserved among TR LBD isoforms to increaseTR-specific selectivity. Conserved structural features of a TR LBD include residues found in equivalent positions of TR LBD isoforms which interact with a conserved structural feature of a compound comprising the biphenyl scaffold (.phi.-X-.phi.) or asingle phenyl scaffold (.phi.-X or X-.phi.) of Formula I. Conformationally constrained structural features of a TR LBD include residues that have their natural flexible conformations fixed by various geometric and physical-chemical constraints, such aslocal backbone, local side chain, and topological constraints. These types of constraints are exploited to restrict positioning of atoms involved in receptor-ligand recognition and binding. For example, comparison of atomic models of TR LBD isoformsbound to thyronine and thyronine-like ligands reveal that certain residues which contact the ligands are restricted to particular topological shapes and angles of rotation about bonds. These include Met259, Leu276, Leu292, His381, Gly290, Ile221, andPhe401 of TR-.alpha.. The corresponding positions in TR-.beta. include Met313, Leu330, Leu346, His435, Gly344, Ile275 and Phe455, respectively.

Selectivity imparted by conformationally constrained features of both the receptor and compound are of particular interest. For example, compounds of Formula I comprising constrained cyclic carbons and substituent groups that interact with aconstrained feature of a TR LBD can be exploited to further increase binding specificity while reducing the potential for cross-over interaction with other receptors. These include hydrophobic and/or hydrophilic contacts between constrained residues ofa TR LBD and atomic groups of the following constituents of the compound in reference to Formula I: (i) the biphenyl rings; (ii) the R3-substituent; (iii) the R3'-substituent; and (iv) the R4'-substituent.

For example, contacts to the phenyl moiety comprising the R1, R2, R3, R5 and R6 substituents, i.e., the ring proximal to the polar pocket (the "inner ring"), include a cycle carbon atom that interacts with an atom of a hydrophobic residue of a TRLBD, such as a carbon and oxygen atom of Met259 and a carbon atom of Leu276 of TR-.alpha., or Met313 and Leu330 of TR-.beta., where the cycle carbon is about 3.0 to 4.0A from the atom of the hydrophobic group. For example, comparison of TR-.alpha. complexed with T3 and TR-.beta. complexed with GC-1 reveals the following conserved inner ring contacts:

Ligand TR LBD T3/Atom TR-.alpha. Residue Atom Distance C11 Met259 C 3.95 C11 Met259 O 3.59 C11 Met259 CB 3.77 C7 Leu276 CD2 3.80 C9 Leu276 CD2 3.70 GC1/Atom TR-.beta. Residue Atom Distance C11 Met313 C 3.85 C11 Met313 O 3.41 C11 Met313CB 3.79 C7 Leu330 CD2 3.56 C9 Leu330 CD2 3.63

Contacts to the phenyl moiety comprising the R2', R3', R4', R5' and R6' substituents, i.e., the ring distal to the polar pocket (the "outer ring"), include a cyclic carbon atom that interacts with an atom of a hydrophobic residue of a TR LBD,such as a carbon atom of Leu292 of TR-.alpha., or Leu346 of TR-.beta., where the cyclic carbon atom is about 3.0 to 4.0A from the atom of the hydrophobic residue. For example, comparison of TR-.alpha. complexed with T3 and TR-.beta. complexed withGC-1 reveals the following conserved outer ring contacts:

Ligand TR LBD T3/Atom TR-.alpha. Residue Atom Distance C6 Leu292 CD2 3.58 C8 Leu292 CD2 3.50 GC1/Atom TR-.beta. Residue Atom Distance C6 Leu346 CD2 3.77 C8 Leu346 CD2 3.80

Contacts to the R3-substituent include an atom that interacts with a carbon atom of a hydrophobic residue of a TR LBD, such as Ile221 of TR-.alpha., or Ile275 of TR-.beta., where the R3-substituent atom is about 3.0 to 4.0A from the carbon atomof the hydrophobic residue. For example, comparison of TR-.alpha. complexed with T3 and TR-.beta. complexed with GC-1 reveals the following conserved R3-substituent contacts:

Ligand TR LBD T3/Atom TR-.alpha. Residue Atom Distance I1 Ile221 CG1 4.01 GC1/Atom TR-.beta. Residue Atom Distance C19 Ile275 CG1 3.98

Contacts to the R3'-substituent include an atom that interacts with an atom of a hydrophobic or hydrophilic residue of a TR LBD, such as an oxygen atom of Gly290 of TR-.alpha., or Gly344 of TR-.beta., where the R3'-substituent atom is about 3.0to 4.0A from the atom of the hydrophobic or hydrophilic residue. For example, comparison of TR-.alpha. complexed with T3 and TR-.beta. complexed with GC-1 reveals the following conserved R4'-substituent, phenolic hydroxyl contacts:

Ligand TR LBD T3/Atom TR-.alpha. Residue Atom Distance I2 Gly290 O 3.50 GC1/Atom TR-.beta. Residue Atom Distance C18 Gly344 O 3.60

Contacts to the R4'-substituent comprising a phenolic hydroxyl include carbon and oxygen atoms that interact with a hydrophobic or hydrophilic residue of a TR LBD, such as a carbon and nitrogen atom of His381 of TR-.alpha., or His435 ofTR-.beta., where the R4'-substituent atom is about 2.0 to 4.0A from an atom of the hydrophobic or hydrophilic residue. For example, comparison of TR-.alpha. complexed with T3 and TR-.beta. complexed with GC-1 reveals the following conservedR4'-substituent, phenolic hydroxyl contacts:

Ligand TR LBD T3/Atom TR-.alpha. Residue Atom Distance C10 His381 CD2 3.97 O1 His381 CD2 3.39 O1 His381 CE1 3.82 C8 His381 NE2 3.47 C10 His381 NE2 3.55 O1 His381 NE2 2.70 GC1/Atom TR-.beta. Residue Atom Distance C10 His435 CD2 3.89 O1His435 CD2 3.64 O1 His435 CE1 3.79 C8 His435 NE2 3.44 C10 His435 NE2 3.33 O1 His435 NE2 2.77

Contacts to the R4'-substituent also may include an atom that interacts with a carbon atom of a hydrophobic residue of a TR LBD, such as Phe401 of TR-.alpha., or Phe455 of TR-.beta., for defining agonist activity, i.e., proper presentation ofhelix-12 (H12) of the TR LBD following ligand binding. The R4'-substituent atom is about 3.0 to 4.0A from the carbon atom of the hydrophobic group. For example, comparison of TR-.alpha. complexed with T3 and TR-.beta. complexed with GC-1 reveals thefollowing conserved R4'-substituent contacts:

Ligand TR LBD T3/Atom TR-.alpha. Residue Atom Distance O1 Phe401 CE1 3.52 O1 Phe401 CZ 3.32 GC1/Atom TR-.beta. Residue Atom Distance O1 Phe455 CE1 3.40 O1 Phe455 CZ 3.22

Comparison of atomic models of TR LBD isoforms complexed with the same and/or different ligands therefore facilitates the identification of new compounds that fit spacially and preferentially into a TR LBD. Modeling, comparison of TR-ligandoverlays, and comparison of TR LBD isoforms also permit identification of conformationally conserved structural features of TR LBD/ligand contacts. Exploiting conformational constraints of the LBD-ligand interaction identified by such methods thereforeimproves the design and identification of new compounds having increased selectivity for binding a particular type of nuclear receptor, such as TR.

TR-.alpha. AND TR-.beta. SELECTIVITY FOR THE THYROID HORMONE RECEPTOR

Using the method described herein ligands can be designed that selectively bind to the alpha more than the beta TR or vice versa. The X-ray crystallographic structure of the rat TR-.alpha. LBD provides insight into design of such ligands.

The three dimensional structure reveals that the major difference between the TR-.alpha. and TR.beta. in the ligand binding cavity resides in amino acid Ser 277 (with the side group --CH2OH) in the rat TR-.alpha. and whose correspondingresidue is 331, asparagine (with the side group --CH2CONH2), in the human TR-.beta.. The side chain in human TR-.beta. is larger, charged and has a different hydrogen bonding potential, which would allow the synthesis of compound; that discriminatebetween this difference. The Ser277 (Asn331 in TR-.beta.) forms part of the polar pocket of the TR LBD, indicating that for TR-.alpha. versus TR-.beta. discrimination, ligands can be designed to contain chemical modification of the R1-substituent withreference to Formula I that exploit this difference.

For example, in the complex of TR-.alpha. with Triac, Ser277 does not participate in ligand binding. The absence of a role for Ser277 (Asn331 in beta) is consistent with the equal affinity of Triac for the alpha and beta isoforms, andindirectly supports the contention that alpha/beta selectivity resides in the amino acid substitution Ser277 to Asn331 and its interaction with Arg228. The effect of the amino acid substitution is further evident when the interactions of Asn331 andArg282 in the structures of the TR-.beta. LBD complexed with GC-1 or Triac are compared with those of Ser277 and Arg228 in the TR-.alpha. LBD. In the complex with GC-1, Asn331 forms a hydrogen bond to Arg282, which in turn forms a hydrogen bond withthe carboxylate of GC-1, a pattern that resembles the interactions of Ser277 and Arg228 in the complexes of the TR-.alpha. LBD complexed with T.sub.3 or Triac. However, in the complex of TR-.beta. with Triac, Arg282 rotates away from Asn331 and theligand, instead forming hydrogen bonds to residues Thr287 and Asp291 of H3. Therefore, differences exist between the two isoforms in the conformation of the polar pocket, depending on the nature of the ligand R1-substituent, indicating that certainsubstituents may interact preferentially with the conformation of a given isoform.

Comparing overlays of various ligands bound to the TR-.alpha. versus TR-.beta. LBDs shows the positioning of the ligand to be very similar. Surprisingly, comparison of the volume and area for the TR-.alpha. and TR-.beta. LBDs bound by thesame or different ligands unexpectedly shows that the cubic space or volume available for accommodating ligand binding by the TR-.beta. LBD (645.+-.28.28 .ANG..sup.3) is larger and more flexable than that of the TR-.alpha. LBD (596.25.+-.7.97.ANG..sup.3) (Table 1). The volume of the ligand binding cavity for TR-.alpha. varies over a narrow range of about 8+, with a maximum difference of about 16+. In contrast, the volume of the ligand binding cavity for TR-.beta. differs by nearly 40+between the complexes with GC-1 and Triac. There also is a difference in the volume of the ligand binding cavity when comparing the same ligand bound to TR-.alpha. and TR-.beta.. For example, TR-.alpha. and TR-.beta. complexed with Triac differ inLBD volume by about 36 .ANG..sup.3. Comparison of TR-.alpha. and TR-.beta. bound to Dimit and GC-1, respectively, which ligands have similar volume/area and superpositioned architecture, show that the difference in LBD volume is about 75 .ANG..sup.3. These differences are attributed primarily to variable movement and interaction of side chain groups with ligand substituents of the phenyl moiety (.phi.) of the biphenyl scaffold (.phi.-X-.phi.) located proximal to the polar pocket, e.g.,R1-substituents in reference to Formula I. In contrast, the volume available in the hydrophobic pocket for both the TR-.alpha. and TR-.beta. LBDs is substantially the same. For example, binding of Triac to the TR-.beta. LBD displaces the side chainof Arg 282 providing approximately 60 .ANG..sup.3 in the polar pocket cavity, exposing the polar pocket to bulk solvent exchange. For GC1 bound to the TR-.beta. LBD, approximately 14 .ANG..sup.3 is due to side chain motion of Met310, and approximately44 .ANG..sup.3 is due to side chain motion of Arg320, the combination of which increases the size of the polar pocket in the TR-.beta. LBD. This extra pliability also may explain the absence of ordered water in the polar pocket of TR-.beta. LBD boundto Triac or GC-1, which is in contrast to the ordered water found in the polar pocket of TR-.alpha. LBD bound to Dimit, IpBr2 or T3.

TABLE 1* rTR-.alpha. hTR-.beta. Dimit Triac IpBr2 T3 GC-1 Triac TR LBD 590/456 589/440 601/474 605/472 665/575 625/474 (vol .ANG..sup.3 / area .ANG..sup.2) Ligand 303/314 333/326 326/330 355/346 294/310 333/326 (vol .ANG..sup.3 / area.ANG..sup.2) Complemen- 0.65 0.68 0.66 0.71 0.61 0.67 tarity *TR LBD volume and area are reported in Angstroms measured by GRASP. Complementarity is determined as defined in Lawrence et al., J. Mol. Biol. 234: 946-950 (1993).

Residue Ser277 in TR-.alpha. and the corresponding residue Asn331 of TR-.beta. also contribute to the volumetric differences observed in the polar pockets of these two TR isoforms. And substitution of the Asn331 of hTR-.beta. with serine hasthe affect of modifying ligand binding affinity of TR-.beta. so that it resembles that of TR-.alpha. (See Example 5). Taken together, differences in hydrogen bonding of atoms of the side chain group of. Ser277 in TR-.alpha. and Asp331 in TR-.beta. extending from the equivalent backbone position in these TR LBDs and the more restricted polar pocket of the TR-.alpha. LBD further supports the concept of designing TR LBD isoform-specific ligands having substituents that fit spacially andpreferentially into the polar pocket of either the TR-.alpha. or TR-.beta. LBDs. Exploitation of this difference provides an additional means for computational design of isoform-specific TR agonists and antagonists.

In terms of ligand design, these differences mean that for .beta.-selective ligands, some or all of the following differences should be exploited:

1. The presence of a larger side chain asparagine.

2. The ability of the carbonyl group on the side chain to provide a strong hydrogen bond acceptor.

3. The ability of the amido group on the side chain to provide a two hydrogen bond donors.

4. Adjustment of polarity to reorganize the trapped water in the T3 pocket.

5. Greater size and flexibility of the polar pocket.

In terms of pharmaceutical design, these differences mean that for .alpha.-selective ligands, some or all of the following differences should be exploited:

1. The presence of a smaller side group.

2. The ability of the hydroxyl on the --CH.sub.2 OH side group carbonyl group on the side chain to provide a weak hydrogen donor.

3. Adjustment of polarity to reorganize the trapped water in the T3 pocket.

4. Smaller size and limited flexibility of the polar pocket.

In both cases these differences can be exploited in a number of ways. For example, they can also be used with a software set for construction of novel organic molecules such as LUDI from Biosym-MSI. An example of designing TR-.beta. selectiveligands is increasing the polarity of a ligand substituent located in the polar pocket of a TR LBD through addition of one or more ligand groups having a formal negative charge and/or negative dipole charge that interacts with a formal positive chargeand/or positive dipole charge of a group in the polar pocket of the LBD. This exploits preferential interactions, such as with the additional positive charge contributed by Asn 331 in TR-.beta.. Another example of a TR-.beta. selective ligand is onethat comprises one or more groups which fit spacially into the TR-.beta. LBD polar pocket. This exploits spacial differences between TR LBD isoforms, such as the larger and more flexible polar pocket of TR-.beta..

METHODS OF TREATMENT

The compounds of Formula 1 can be useful in medical treatments and exhibit biological activity which can be demonstrated in the following tests:

(i) the induction of mitochondrial .alpha.-glycerophosphate dehydrogenase (GPDH:EC 1.1.99.5). This assay is particularly useful since in certain species e.g. rats it is induced specifically by thyroid hormones and thyromimetics in aclose-related manner in responsive tissues e.g. liver, kidney and the heart (Westerfield, W. W., Richert, D. A. and Ruegamer, W. R., Endocrinology (1965) 77:802). The assay allows direct measurement in rates of a thyroid hormone-like effect of compoundsand in particular allows measurement of the direct thyroid hormone-like effect on the heart. Other measurements included parameters such as heart rate and cardiac enzymes including Ca.sup.++ ATPase, Na.sup.++ /K.sup.+ ATPase, myosin isoforms andspecific liver enzymes;

(ii) the elevation of basal metabolic rate as measured by the increase in whole body oxygen consumption (see e.g., Barker et al., Ann. N. Y. Acad. Sci., (1960) 86:545-562);

(iii) the stimulation of the rate of beating of atria isolated from animals previously dosed with thyromimetrics (see e.g., Stephan et al., Biochem. Pharmacol. (1992) 13:1969-1974; Yokoyama et al., J. Med. Chem., (1995) 38:695-707);

(iv) the change in total plasma cholesterol levels as determined using a cholesterol oxidase kit (for example, the Merck CHOD iodine colorimetric kit. see also, Stephan et al. (1992));

(v) the measurement of LDL (low density lipoprotein) and HDL (high density lipoprotein) cholesterol in lipoprotein fractions separated by ultracentrifugation; and p (vi) the change in total plasma triglyceride levels as determined using enzymaticcolor tests, for example the Merck System GPO-PAP method.

The compounds of Formula 1 can be found to exhibit selective thyromimetic activity in these tests,

(a) by increasing the metabolic rate of test animals, and raising hepatic GPDH levels at doses which do not significantly modify cardiac GPDH levels.

(b) by lowering plasma cholesterol and triglyceride levels, and the ratio of LDL to HDL cholesterol at doses which do not significantly modify cardiac GPDH levels.

The compounds of Formula 1 may therefore be used in therapy, in the treatment of conditions which can be alleviated by compounds which selectively mimic the effects of thyroid hormones in certain tissues whilst having little or no directthyromimetic effect on the heart. For example, compounds of Formula 1 which raise hepatic GPDH levels and metabolic rate at doses which do not significantly modify cardiac GPDH levels are indicated in the treatment of obesity.

Agonists of Formula 1 will lower total plasma cholesterol, the ratio of LDL-cholesterol to HDL-cholesterol and triglyceride levels at doses which do not significantly modify cardiac GPDH levels are indicated for use as general antihyperlipidaemic(antihyperlipoproteinaemic) agents i.e. in the treatment of patients having elevated plasma lipid (cholesterol and triglyceride) levels. In addition, in view of this effect on plasma cholesterol and triglyceride, they are also indicated for use asspecific anti-hypercholesterolemic and anti-hypertriglyceridaemic agents.

Patients having elevated plasma lipid levels are considered at risk of developing coronary heart disease or other manifestations of atherosclerosis as a result of their high plasma cholesterol and/or triglyceride concentrations. Further, sinceLDL-cholesterol is believed to be the lipoprotein which induces atherosclerosis, and HDL-cholesterol believed to transport cholesterol from blood vessel walls to the liver and to prevent the build up of atherosclerotic plaque, anti-hyperlipidemic agentswhich lower the ratio of LDL-cholesterol to HDL cholesterol are indicated as anti-atherosclerotic agents, herein incorporated by reference U.S. Pat. Nos. 4,826,876 and 5,466,861.

The present invention also provides a method of producing selective thyromimetic activity in certain tissues except the heart which comprises administering to an animal in need thereof an effective amount to produce said activity of a compound ofFormula 1 or a pharmaceutically acceptable salt thereof.

The present invention also relates to a method of lowering plasma lipid levels and a method of lowering the ratio of LDL-cholesterol to HDL-cholesterol levels by suitably administering a compound of this invention or a pharmaceutically acceptablesale thereof.

In addition, compounds of Formula 1 may be indicated in thyroid hormone replacement therapy in patients with compromised cardiac function.

In therapeutic use the compounds of the present invention are usually administered in a standard pharmaceutical composition.

The present invention therefore provides in a further aspect pharmaceutical compositions comprising a compound of Formula 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Such compositions include thosesuitable for oral, parenteral or rectal administration.

PHARMACEUTICAL COMPOSITIONS

Compounds of Formula 1 and their pharmaceutically acceptable salts which are active when given orally can be formulated as liquids for example syrups, suspensions or emulsions, tablets, capsules and lozenges.

A liquid composition will generally consist of a suspension or solution of the compound or pharmaceutically acceptable salt in a suitable liquid carrier(s), for example ethanol, glycerine, sorbitol, non-aqueous solvent such as polyethyleneglycol, oils or water, with a suspending agent, preservative, surfactant, wetting agent, flavoring or coloring agent. Alternatively, a liquid formulation can be prepared from a reconstitutable powder.

For example a powder containing active compound, suspending agent, sucrose and a sweetener can be reconstituted with water to form a suspension; and a syrup can be prepared from a powder containing active ingredient, sucrose and a sweetener.

A composition in the form of a tablet can be prepared using any suitable pharmaceutical carrier(s) routinely used for preparing solid compositions. Examples of such carriers include magnesium stearate, starch, lactose, sucrose, microcrystallinecellulose and binders, for example polyvinylpyrrolidone. The tablet can also be provided with a color film coating, or color included as part of the carrier(s). In addition, active compound can be formulated in a controlled release dosage form as atablet comprising a hydrophilic or hydrophobic matrix.

A composition in the form of a capsule can be prepared using routine encapsulation procedures, for example by incorporation of active compound and excipients into a hard gelatin capsule. Alternatively, a semi-solid matrix of active compound andhigh molecular weight polyethylene glycol can be prepared and filled into a hard gelatin capsule; or a solution of active compound in polyethylene glycol or a suspension in edible oil, for example liquid paraffin or fractionated coconut oil can beprepared and filled into a soft gelatin capsule. Compound of Formula 1 and their pharmaceutically acceptable salts which are active when given parenterally can be formulated for intramuscular or intravenous administration.

A typical composition for intramuscular administration will consist of a suspension or solution of active ingredient in an oil, for example arachis oil or sesame oil. A typical composition for intravenous administration will consist of a sterileisotonic aqueous solution containing, for example active ingredient, dextrose, sodium chloride, a co-solvent, for example polyethylene glycol and, optionally, a chelating agent, for example ethylenediamine tetracetic acid and an anti-oxidant, forexample, sodium metabisulphite. Alternatively, the solution can be freeze dried and then reconstituted with a suitable solvent just prior to administration.

Compounds of structure (1) and their pharmaceutically acceptable salts which are active on rectal administration can be formulated as suppositories. A typical suppository formulation will generally consist of active ingredient with a bindingand/or lubricating agent such as a gelatin or cocoa butter or other low melting vegetable or synthetic wax or fat.

Compounds of Formula 1 and their pharmaceutically acceptable salts which are active on topical administration can be formulated as transdermal compositions. Such compositions include, for example, a backing, active compound reservoir, a controlmembrane, liner and contact adhesive.

The typical daily dose of a compound of Formula 1 varies according to individual needs, the condition to be treated and with the route of administration. Suitable doses are in the general range of from 0.001 to 10 mg/kg bodyweight of therecipient per day.

Within this general dosage range, doses can be chosen at which the compounds of Formula 1 lower plasma cholesterol levels and raise metabolic rate with little or no direct effect on the heart. In general, but not exclusively, such doses will bein the range of from lower doses (0.001 to 0.5 mg/kg) to higher doses (0.5 to 10 mg/kg).

In addition, within the general dose range, doses can be chosen at which the compounds of Formula 1 lower plasma cholesterol levels and have little or no effect on the heart without raising metabolic rate. In general, but not exclusively, suchdoses will be in the range of from 0.001 to 0.5 mg/kg.

It is to be understood that the 2 sub ranges noted above are not mutually exclusive and that the particular activity encountered at a particular dose will depend on the nature of the compound of Formula 1 used.

Preferably, the compound of Formula 1 is in unit dosage form, for example, a tablet or a capsule so that the patient may self-administer a single dose. In general, unit doses contain in the range of from 0.05-100 mg of a compound of Formula 1. Preferred unit doses contain from 0.05 to 10 mg of a compound of Formula 1.

The active ingredient may be administered from 1 to 6 times a day. Thus daily doses are in general in the range of from 0.05 to 600 mg per day. Preferably, daily doses are in the range of from 0.05 to 100 mg per day. Most preferably from 0.05to 5 mg per day.

EXAMPLES

EXAMPLE 1--SYNTHESIS OF TR LIGANDS

Many TR ligands are known in the art, including T4 (thyroxine), T3, T2 and TS-9. See Jorgensen, Thyroid Hormones and Analogs, in Hormonal Proteins and Peptides, Thyroid Hormones 107-204 (Choh Hao Li ed., 1978), incorporated by reference herein.

The syntheses of several TR ligands are described below.

Synthesis of TS1, TS2, TS3, TS4, TS5

TS1, TS2, TS3, TS4 and TS5 and analogs thereof can all be prepared by simple acylation of the nitrogen atom of any thyronine analog, including T3 (3,5,3'-triiodo-L-thyronine), T4 (thyroxine) and 3,5-diiodothyronine. TS1 and TS2 are synthesizedby reacting T3 with Ph.sub.2 CHCO.sub.2 NHS (N-hydroxy succinimide-2,2-diphenylacetate) and C.sub.16 H.sub.33 CO.sub.2 NHS, respectively. TS3 is synthesized by reacting T3 with FMOC-Cl (fluorenylmethyloxycarbonylchloride). TS4 is synthesized byreacting T3 with tBOC.sub.2 O (tBOC anhydride or di-t-butyldicarbonate). TS5, which differs from TS1-4 by having a --H instead of an --I at the R.sub.3.sup.1 position, is synthesized by reacting 3,5-diiodothyronine with tBOC.sub.2 O. The generalreaction scheme for TS1, TS2, TS3, TS4 and TS5 is depicted in FIG. 11. It should be noted that in the reaction scheme, both TS5 and its precursor both have a hydrogen rather than an iodine at the R.sub.3.sup.1 position.

Synthesis of TS6 and TS7

TS6 is synthesized by reacting TS5 with paranitrophenylisocyanate. TS7 is synthesized by reacting TS6 with TFA (trifluoroacetic acid), which cleaves the tBOC group. These reactions are simple organic synthesis reactions that can be performed byanyone of ordinary skill in the art. The synthetic scheme for TS6 and TS7 is diagrammed in FIG. 12.

Synthesis of TS8

TS8 is synthesized by reacting TS5 with Ph.sub.2 CHNH.sub.2 (diphenylmethylamine) in the presence of triethylamine and any amide forming condensing reagent, such as TBTU (hydroxybenztriazoleuronium tetrafluoroborate) or HBTU(hydroxybenztriazoleuronium hexafluorophosphate). The synthesis scheme for TS8 is depicted in FIG. 13.

SYNTHESIS OF 3,5-DIIODO-3'ISOPROPYLTHYRONINE DERIVATIVES

For designing a class of antagonists, it is important to have a hydrophobic group at the 3' position as well as an extension at the 5' position. Preferred hydrophobic groups at the 3' position include: methyl, benzyl, phenyl, iodo, andheterocyclic structures. The synthesis of a 3,5-diiodo-3'-isopropyl-5'-substituted thyronine is described below. The example provided describes the specific steps for synthesizing the TS10 compound, but this general reaction scheme can be used by oneof ordinary skill in the art to synthesize any number of 3,5, -diiodo-3'-isopropyl-5'-substituted thyronine derivatives, which are characterized by having an extension at the 5' position. Additional compounds of this class can be synthesized using knownorganic synthesis techniques.

The synthesis of TS10 is described below and is depicted in FIG. 14. Numbers used in the reaction scheme for TS10 indicating the reaction product for each step are in parentheses.

2-Formyl-6-isopropylanisole (1): 2-formyl-6-isopropylanisole (10.0 g, 61 mmol), as made by Casiraghi, et al. JCS Perkin I, 1862 (1980) (incorporated by reference), is added dropwise to a suspension of sodium hydride (3.7 g, 153 mmol) in 50 mL THFand 50 mL of DMF in a round bottom flask. The addition generates an exothermic reaction and formation of a gray solid. Methyl iodide (26.0 g, 183 mmol) is then added dropwise and the reaction mixture is stirred at room temperature for 5 hours. Thereaction mixture is quenched with 20 mL of water, then poured into 500 mL of water, and is extracted with ether (2.times.300 mL). The ether layers are combined, washed with water (5.times.1000 mL), dried over magnesium sulfate and concentrated in vacuoto provide 10.2 g (94%) of the title compound, with the following .sup.1 H NMR (CDCl.sub.3) properties: d 10.30 (s, 1H), 7.63 (d, 1H, J=3 Hz), 7.50 (d, 1H, J=3 Hz), 7.13 (t, 1H, J=3 Hz), 3.81 (s, 3H), 3.31 (heptet, 1H, J=7.5 Hz), 1.19 (d, 6H, J=7.5 Hz).

2-(2-Hydroxynonyl)-6-isopropylanisole (not shown in scheme): Octylmagnesium chloride (8.4 mL, 16.9 mmol, 2.0 M) is added dropwise to a solution of 1 (1.5 g, 8.4 mmol) in 10 mL THF at -78.degree. C. The reaction mixture is stirred for 2 hourswith warming to room temperature. The reaction mixture is diluted with 50 mL ether and poured into 50 mL water. The ether layer is washed with brine (1.times.50 mL), dried over sodium sulfate, and concentrated in vacuo. Flash chromatography (silicagel, 10% ether/hexane.fwdarw.15% ether/hexane) provides 734 mg (30%) of the title compound with the following .sup.1 H NMR (CDCl.sub.3) properties: d 7.33-7.10 (m, 3H), 5.00 (br. s, 1H), 3.81 (s, 3H), 3.33 (heptet, 1H, J=7 Hz) 1.90-1.19 (m, 14H), 0.86(t, 3H, J=6.5 Hz); HRMS (EI), found: 292.2404; calc'd: 292.2402.

2-nonyl-6-isopropylanisole (2): Compound 2 (663 mg, 2.3 mmol) is dissolved in solution 5 mL ethanol and 5 mL acetic acid, and a spatula tip of palladium on carbon catalyst is added. The reaction mixture is then charged with hydrogen gas (using asimple balloon and needle) and the mixture is stirred at room temperature overnight. The next day, the reaction mixture is poured into ether (100 mL) and the ether layer is extracted with saturated sodium bicarbonate (3.times.100 mL). The ether layeris dried over sodium sulfate and concentrated in vacuo to provide 581 mg (91%) of (2) with the following .sup.1 H NMR (CDCl.sub.3) properties: d 7.14-7.00 (m, 3H), 3.75 (s, 3H), 3.36 (heptet, 1H, J=6.8 Hz), 2.63 (t, 2H, J=7.5 Hz), 1.68-1.15 (m, 14H),0.86 (t, 3H, J=5.5 Hz); HRMS (EI), mass found: 276.2459; calculated: 276.2453.

Thyronine adduct (4): Fuming nitric acid (0.071 mL) is added to 0.184 mL acetic anhydride chilled to -5.degree. C. Iodine (66 mg) is added to this mixture followed by trifluoroacetic acid (0.124 mL). This mixture is stirred for 1 hour withwarming to room temperature, at which point all of the iodine is dissolved. The reaction mixture was then concentrated in vacuo to provide an oily semi-solid material. The residue was dissolved in 0.7 mL of acetic anhydride and cooled to -20.degree. C. A solution of anisole (2) (581 mg, 2.1 mmol) in 1.2 mL acetic anhydride and 0.58 mL TFA is added dropwise. The reaction mixture is stirred at -20.degree. for 1 hour, then stirred overnight with warming to room temperature. The reaction mixture ispartitioned between water and methylene chloride. The methylene chloride layer is dried over sodium sulfate and concentrated in vacuo to provide the iodonium salt (3) as an oil. This material is not purified or characterized, and is directly introducedinto the coupling reaction.

N-Trifluoroacetyl-3,5-diiodotyrosine methyl ester (552 mg, 1.0 mmol) prepared according to the procedure of N. Lewis and P. Wallbank, Synthesis 1103 (1987) (incorporated by reference) and all of the crude iodonium salt (3) from above is dissolvedin 5 mL of anhydrous methanol. Diazabicyclo[5.4.0]undecane (DBU) (183 mg, 1.2 mmol) and a spatula tip of copper-bronze are added and the resulting mixture is stirred at room temperature overnight. The next day, the reaction mixture is filtered, and thefiltrate is concentrated in vacuo. The crude residue is purified by flash chromatography (silica gel, 10% ethyl acetate/hexane) to provide 30 mg (4%) of the protected thyronine adduct (4).

Deprotected thyronine (TS10): The protected thyronine 4 (30 mg, 0.04 mmol) is dissolved in a mixture of 2.25 mL acetic acid and 2.25 mL 49% hydrobromic acid. The reaction mixture is heated to reflux for 5 hours. The reaction mixture is cooledto room temperature, and the solvents are removed in vacuo. Water is added to triturate the oily residue into a gray solid. This solid material is filtered, washed with water, and dried over P.sub.2 O.sub.5 in vacuo to provide 24 mg (81%) of the tidecompound, TS10, with the following .sup.1 H NMR (CDCl.sub.3) properties: d 7.57 (s, 1H), 6.86 (s, 1H), 6.45 (s, 1H), 6.34 (s, 1H), 4.81 (m, 1H), 3.86 (s, 3H), 3.71 (s, 3H), 3.33-3.05 (m, 3H), 2.58-2.47 (m, 2H), 1.62-0.76 (m, 23H); MS (LSIMS): M.sup.+=817.0.

As mentioned above, this reaction scheme can be modified by one of ordinary skill in the art to synthesize a class of compounds characterized by 3,5-diiodo-3' isopropylthyronine derivatives, wherein (1) the 3' isopropyl group can be replaced witha hydrophobic group, including methyl, benzyl, phenyl, iodo, and heterocyclic structures, and (2) a wide variety of chemical structures can be incorporated at the 5' position, including alkyl groups, planar aryl, heterocyclic groups, or polar and/orcharged groups.

The aldehyde (1) in the above reaction scheme is a versatile synthetic intermediate which allows for the attachment of a variety of chemical moieties to the 5' position of the final thyronine derivative. In addition, a variety of chemicalreactions can be used to attach the chemical moieties. These reactions are well known in the art and include organometallic additions to the aldehyde (including Grignard reagents, organolithiums, etc.), reductive amination reactions of the aldehyde witha primary or secondary amine, and Wittig olefination reactions with a phosphorous ylid or stabilized phosphonate anion. Other possibilities include reduction of the aldehyde to a benzyl alcohol allowing for etherification reactions at the 5' position. As mentioned above, these methods allow for a wide variety of chemical structures to be incorporated at the 5' position of the final thyronine derivative, including alkyl groups, planar aryl, heterocyclic groups or polar and/or charged groups.

Synthesis of 3,5-dibromo-4-(3',5'-diisopropyl-4'-hydroxyphenoxy)benzoic acid (Compound 11). ##STR3##

(a) A mixture of 2,6-diisopropyl phenol (20 g, 0.11 mol), potassium carbonate (62 g, 0.45 mol), acetone (160 ml) and methyl iodide (28 ml, 0.45 mole) is refluxed for three days. The reaction mixture is filtered through celite, evaporated,dissolved in ether, washed twice with 1M sodium hydroxide, dried over magnesium sulphate and concentrated to afford 15.1 g (0.08 mol, 70%) of 2,6-diisopropyl anisole as a slightly yellow oil.

(b) Fuming nitric acid (12.4 ml, 265 mmol) is added dropwise to 31.4 ml of acetic anhydride which is cooled in a dry ice/carbon tetrachloride bath. Iodine 11.3 g, 44.4 mmol) is added in one portion followed by dropwise addition oftrifluoroacetic acid (20.5 ml, 266 mmole). The reaction mixture is stirred at room temperature until all the iodine is dissolved. Nitrogen oxides are removed by flushing nitrogen into the vessel. The reaction mixture is concentrated, the residue isdissolved in 126 ml of acetic anhydride and is cooled in a dry ice/carbon tetrachloride bath. To the stirred solution 2,6-diisopropylanisole (51 g, 266 mmol) in 150 ml of acetic anhydride and 22.6 ml of trifluoroacetic acid is added dropwise. Thereaction mixture is left to stand at room temperature over night and then is concentrated. The residue is taken up in 150 ml of methanol and treated with 150 ml of 10% aqueous sodium bisulfite solution and 1 liter of 2M sodium borotetrafluoridesolution. After the precipitate aggregates, petroleum ether is added and the supernatant is decanted. The precipitate is triturated with petroleum ether, filtered, washed with petroleum ether and dried at room temperature in vacuo. This affords 34 g(57 mmol, 65%) of bis(3,5-diisopropyl-4-methoxyphenyl)iodonium tetrafluoroborate as a white solid.

(c) To a stirred solution of 3,5-dibromo4-hydroxybenzoic acid (12 g, 40.5 mmol) in 250 ml of methanol, thionyl chloride (3 ml) is added dropwise. The reaction mixture is refluxed for five days, water is added and the precipitated product isfiltered off. The residue is dissolved in ethyl acetate. From the aqueous phase, methanol is removed by concentration. The aqueous phase is then saturated with sodium chloride, and extracted with ethyl acetate. The combined organic phases are driedover magnesium sulphate, filtered and concentrated. This gives 12.5 g (40.5 mmol, 100%) of 3,5-dibromo-4-hydroxymethyl benzoate as a white crystalline solid.

(d) The products obtained in steps b and c are reacted with each other according to the following protocol. To bis(3,5-diisopropyl-4-methoxyphenyl)iodonium tetrafluoroborate (2.86 g, 4.8 mmole) and copper bronze (0.42 g, 6.4 mmole) in 7 ml. ofdichloromethane at 0.degree. C. is added dropwise a solution of 3,5-dibromo-4-hydroxymethyl benzoate (1.0 g, 3.2 mmole) and triethylamine (0.36 g, 3.5 mmole) in 5 ml of dichloromethane. The reaction mixture is stirred in the dark for eight days andthen is filtered through celite. The filtrate is concentrated and the residue is purified by column chromatography (silica gel, 97:3 petroleum ether/ethyl acetate) to give 0.62 g (1.2 mmole, 39%) of3,5-dibromo-4-(3',5'-diisopropyl-4'-methoxyphenoxy)methyl benzoate as a solid.

(e) The product from step d (0.2 g, 0.4 mmole) is dissolved in 2 ml. dichloromethane, is put under nitrogen and is cooled at -40.degree. C. To the stirred solution is added 1M BBr.sub.3 (1.2 ml, 1.2 mmole) dropwise. The reaction mixture isallowed to reach room temperature and then is left over night. It is cooled to 0.degree. C. and then hydrolyzed with water. Dichloromethane is removed by concentration and the aqueous phase is extracted with ethyl acetate. The organic phase is washedwith 1M hydrochloric acid and brine. Then it is dried over magnesium sulphate, filtered and concentrated. The residue is chromatographed (silica, 96:3.6:0.4 dichloromethane/methanol/acetic acid) producing 93 mg (0.2 mmole, 51%) of3,5-dibromo-4-(3',5'-diisopropyl-4'-hydroxyphenoxy)benzoic acid as a white solid. .sup.1 H nmr (CDCl.sub.3) .delta. 1.23 (d, 12H, methyl), 3.11 (m, 2H, CH), 6.50 (s, 2H, 2,6-H) 8.33 (s, 2H, 2',6'-H).

Synthesis of addition ligands are described in U.S. Ser. No. 08/877,792, filed Jun. 18, 1997 which is herein incorporated in its entirety by reference.

TABLE 2 and FIG. 15 depict the structures of several TR ligands in reference to Formula I.

TABLE 2 Cmpd R.sub.3 R.sub.4 R.sub.5 R'.sub.3 R'.sub.4 R'.sub.5 R.sub.1 *T.sub.3 --I --O-- --I --I --OH --H --CH.sub.2 CH(NH.sub.2)CO.sub.2 H *T.sub.4 --I --O-- --I --I --OH --I --CH.sub.2 CH(NH.sub.2)CO.sub.2 H TS1 --I --O-- --I --I --OH--H --CH.sub.2 CH[NHCOCH.phi..sub.2 ]CO.sub.2 H TS2 --I --O-- --I --I --OH --H --CH.sub.2 CH[NHCO(CH.sub.2).sub.15 CH.sub.3 ]CO.sub.2 H TS3 --I --O-- --I --I --OH --H --CH.sub.2 CH[NH-FMOC]CO.sub.2 H TS4 --I --O-- --I --I --OH --H --CH.sub.2 CH[NH-tBOC]CO.sub.2 H TS5 --I --O-- --I --H --OH --H --CH.sub.2 CH[NH-tBOC]CO.sub.2 H TS6 --I --O-- --I --H --OC(O)NH.dbd..0..sub.p NO.sub.2 --H --CH.sub.2 CH[NH-tBOC]CO.sub.2 H TS7 --I --O-- --I --I --OC(O)NH.dbd.NH.0.NO.sub.2 --H --CH.sub.2CH(NH.sub.2)CO.sub.2 H TS8 --I --O-- --I --H --NH--CH.0..0. --H --CH.sub.2 CH[NH-tBOC]CO.sub.2 H TS9 --I --O-- --I -IsoPr --OH --H --CH.sub.2 CH(NH.sub.2)CO.sub.2 H TS10 --I --O-- --I -IsoPr --OH --(CH).sub.8 -- --CH.sub.2 CH(NH.sub.2)CO.sub.2 H CH.sub.3 *Prior Art Compound --.0.: phenyl --.0.OpNO.sub.2 : para nitro phenyl

EXAMPLE 2--RECEPTOR BINDING ASSAYS OF TR LIGANDS

To test the ability of synthesized TR ligands to bind to a thyroid receptor (TR), the binding affinity of a TR ligand for TR is assayed using TR's prepared from rat liver nuclei and 125.sub.I T.sub.3 as described in J. D. Apriletti, J. B. Baxter,and T. N. Lavin, J. Biol. Chem., 263: 9409-9417 (1988). The apparent Kd's are calculated using the method described by Apriletti (1995) and Apriletti (1988). The apparent Kd's are presented in TABLE 3. The apparent Kd's (App.Kd) are determined in thepresence of the sample to be assayed, 1 nM [.sup.125 I]T.sub.3, and 50 .mu.g/ml core histones, in buffer E (400 mM KCl, 200 mM potassium phosphate, pH 8.0, 0.5 mM EDTA, 1 mM MgCl.sub.2, 10% glycerol, 1 mM DTT) in a volume of 0.21 ml. After incubationovernight at 4.degree. C., 0.2 ml of the incubation mixture is loaded onto a Quick-Sep Sephadex G-25 column (2.7.times.0.9 cm, 1.7 ml bed volume) equilibrated with buffer E. The excluded peak of protein-bound [.sup.125 I]T.sub.3 is eluted with 1 ml ofbuffer E, collected in a test tube, and counted. Specific T.sub.3 binding is calculated by subtracting nonspecific binding from total binding.

TABLE 3 Coactivation Assay Compound App.Kd (nM) RIP-140 EC.sub.50 (M) T.sub.3 0.06 + .sup. 10.sup.-10 T.sub.4 2 + 10.sup.-9 TS1 4 + 10.sup.-7 TS2 1400 nd nd TS3 4 + 10.sup.-8 TS4 8 + nd TS5 220 + 10.sup.-6 TS6 >10000 nd nd TS7 260+ 10.sup.-7 TS8 6000 nd nd TS9 1 + .sup. 10.sup.-10 TS10 400 + 10.sup.-6 +: RIP-140 Binding -: RIP-140 Binding nd: Not Determined

EXAMPLE 3--INCREASED NUCLEAR PROTEIN COACTIVATION BY TR LIGANDS

To test the ability of TR ligands to activate the binding of TR to the nuclear activation protein RIP-140 (a nuclear protein that can bind to nuclear receptors, such as the estrogen receptor), a TR ligand is liganded to TR and then incubated withRIP-140 as described in V. Cavailles, et al., EMBO J., 14(15):3741-3751 (1995), which is incorporated by reference herein. In this assay, 35.sub.s -RIP-140 protein binds to liganded TR but not unliganded TR. Many TR 35.sub.s ligands can activateRIP-140 binding as shown in TABLE 3.

EXAMPLE 4--TR LIGAND BINDING AND TR ACTIVATION IN CULTURED CELLS

To test TR activation of transcription in a cellular environment, TR ligands are assayed for their ability to activate a reporter gene, chloramphenicol transferase ("CAT"), which has a TR DNA binding sequence operatively linked to it. Either GCor L937 cells (available from the ATCC) can be used, respectively). In such assays, a TR ligand crosses the cell membrane, binds to the TR, and activates the TR, which in turn activates gene transcription of the CAT by binding the TR DNA binding regionupstream of the CAT gene. The effective concentration for half maximal gene activation (EC.sub.50) is determined by assaying CAT gene activation at various concentrations as described herein and in the literature. The results of CAT gene activationexperiments are shown in TABLE 3.

CAT GENE ACTIVATION ASSAYS

Functional response to thyroid hormone (3,5,3'-triiodo-L-thyronine, T.sub.3) and TR ligands is assessed either in a rat pituitary cell line, GC cells, that contain endogenous thyroid hormone receptors (TRs) or U937 cells that contain exogenousTRs expressed as known in the art. GC cells are grown in 10-cm dishes in RPMI 1640 with 10% newborn bovine serum, 2 mM glutamine, 50 units/ml penicillin and 50 .mu.g/ml streptomycin. For transfections, cells are trypsinized, resuspended in buffer (PBS,0.1% glucose) and mixed with a TREtkCAT plasmid (10 mg) or phage in 0.5 ml buffer (15.+-.5 million cells) and electroporated using a Bio-Rad gene pulser at 0.33 kvolts and 960 mF. The TREtkCAT plasmid contains two copies of a T.sub.3 response element(AGGTCAcaggAGGTCA) cloned in the Hind III site of the pUC19 polylinker immediately upstream of a minimal (-32/+45) thymidine kinase promoter linked to CAT (tkCAT) coding sequences. After electroporation, cells are pooled in growth medium (RPMI with 10%charcoal-treated, hormone stripped, newborn bovine serum), plated in 6-well dishes and treated with either ethanol or hormone. CAT activity is determined 24 hours later as described D. C. Leitman, R. C. J. Ribeiro, E. R. Mackow, J. D. Baxter, B. L.West, J. Biol. Chem. 266, 9343 (1991), which is incorporated by reference herein.

EFFECT OF TS-10 ON THE TRANSCRIPTIONAL REGULATION OF THE DR4-ALP REPORTER GENE IN THE PRESENCE OR ABSENCE OF T3.

Characteristics of the TRAF cells: TRAFa1 are CHO K1 cells stably transformed with an expression vector encoding the human thyroid hormone receptor .alpha. 1 and a DR4,ALP reporter vector; TRAFb1 are CHO K1 cells stably transformed with anexpression vector encoding the human thyroid hormone receptor .beta.1 and a DR4-ALP reporter vector.

Interpretation of the Effect of Compound TS-10 on the Transcriptional Regulation of the DR4-ALP Reporter Gene in the Presence or Absence of T3.

TRAFa1 reporter cells: TS-10 alone (open circles) induces a partial activation of the expression of the ALP reporter protein amounting to approximately 27% of the maximal effect by the natural thyroid hormone T3. In the presence of T3 (filledcircles), TS-10 has a weak antagonistic effect. The EC50 concentration for the agonistic effect of TS-10 and the EC50 concentration for its T3 antagonistic effect, respectively, is indicated in FIG. 18.

In FIG. 18, open and filled circles with dotted lines show the dose-dependent effect of TS-10/T3 on the toxicity marker (MTS/PMS), reduction of tetrazolium salt in the mitochondria, displayed on the right y-axis as optical density. There is noobvious toxic effect of TS-10 on the MTS-PMS marker but there is a clear effect on the morphology of the cells, as can be seen under the light microscope, at the highest concentration of TS-10 (32 mM) both in the absence and presence of T3, respectively(not shown in the figure).

TRAFb1 reporter cells: TS-10 alone (open circles) induces a partial activation of the expression of the ALP reporter protein amounting to approximately 35% of the maximal effect by 13. The EC50 concentration for the agonistic effect of TS-10 isindicated in FIG. 19. In the presence of T3 (filled circles), TS-10 shows, if anything, a slight potentiation of the T3 effect on the expression of the ALP reporter protein. The T3 inhibitory effect of TS-10 at its highest concentration used (32 mM) isa toxic effect rather than T3 antagonism.

In FIG. 19, open and filled circles with dotted lines show the dose-dependent effect of TS-10/T3 on ihe toxicity marker (MTS/PMS), reduction of tetrazolium salt in the mitochondria, displayed on the right y-axis as optical density. There is noobvious toxic effect of TS-10 on the MTS-PMS marker but a clear effect on the morphology of the cells can be observed, under the light microscope, at the highest concentration of TS-10 (32 mM) both in the absence and presence of T3, respectively (notshown in the figure).

HepG2 (HAF18) reporter cells: TS-10 alone (open circles) induces a partial activation of the expression of the ALP reporter protein amounting to slightly more than 50% of the maximal effect by T3. The EC50 concentration for the agonistic effectof TS-10 is indicated in FIG. 20. In the presence of T3 (filled circles), TS-10 shows no effect i.e. no T3 antagonism nor potentiation/additive effect to T3. Open and filled circles with dotted lines show the dose-dependent effect of TS-10/T3 on thetoxicity marker (MTS/PMS), reduction of tetrazolium salt in the mitochondria, displayed on the right y-axis as optical density. There is no obvious toxic effect of TS-10 on the MTS/PMS marker or on the morphology of the cells, as can be observed using alight microscope, at any concentration of TS-10/T3 used.

EXAMPLE 5--COMPARISONS OF HUMAN TR-.alpha. AND HUMAN TR-.beta.

Competition for [.sup.125 I]T.sub.3 binding to TR LBD by T.sub.3 and Triac

The drug, Triac, is a thyroid hormone agonist. Triac is 3,5,3'-triiodothyroacetic acid and is described in Jorgensen, Thyroid Hormones and Analogs in Hormonal Proteins and Peptides, Thyroid Hormones at 150-151 (1978). Another compound that canbe used in place of Triac is 3,5-diiodo-3'-isopropylthyroacetic acid. Competition assays are performed to compare the displacement of [.sup.125 I]T.sub.3 from binding with human TR-.alpha. LBD or human TR-.beta. LBD by unlabeled T.sub.3 or Triac. Theresults of such assays are depicted in FIG. 16.

Standard binding reactions are prepared containing 1 nM [.sup.125 I]T.sub.3, 30 fmol of human TR-.alpha. (empty symbols) or .beta. (solid symbols), and various concentrations of competing unlabeled T.sub.3 (circles) or Triac (triangles). Assays are performed in duplicate.

Competition for [.sup.125 I]T.sub.3 binding to variant TR LBD by T.sub.3, Triac and GC-1

The following assays residues involved in selective binding among TR isoforms. Competition assays are performed to compare the displacement of [.sup.125 I]T.sub.3 from binding with wild-type human TR-.alpha. LBD or human TR-.beta. LBD, to avariant form of the TR LBDs by unlabeled T.sub.3, Triac or GC-1. A variant TR-.alpha. or TR.beta. is constructed by substituting an amino acid found in the corresponding position of the other TR isoform. For example, asparagine 331 in human TR.beta. corresponds to serine 277 in human TR.alpha.. To test binding specificity contributed by this position, a variant human TR-.beta. is constructed that contains asparagine 331 substituted with a serine residue (designated Asn331Ser or N331S). Bindingassays are described in Apriletti et al. (Protein Expression and Purification 6:363-370 (1995)). The results of such assays are depicted in FIG. 27, and summarized in Table 4 below.

TABLE 4 Effect of TR-.beta. Substitution N331S on Binding Affinity Ligand Native TR-.alpha. Native TR-.beta. Mutant TR-.beta. T3 .sup. 20 pM .sup. 60 pM .sup. 100 pM T4 600 3000 ND Triac 20 20 100 IpBr.sub.2 17 ND ND Dimit 6000 8000 ND GC-1 200 40 400

Competition curves comparing wildtype TR-.beta. versus the variant TR-.beta. N331S for binding T3, Triac or GC-1 show that the affinity of the mutant receptor for Triac was reduced to approximately the same as for T3 (vs. 3-fold greater inwild type) so that the relative affinities are similar to wild-type TR-.alpha.. The affinity for GC-1 was also reduced to several fold less than T3, as is seen with TR-.alpha..

Comparison of the affinity of TR-.beta. variant N331S to the native TRs for selected ligands is as follows:

Native TR-.alpha. for various ligands (T3, T4, Triac, IpBr2, Dimit, GC-1):

IpBr.sub.2 >Triac.apprxeq.T3>GC-1>T4>Dimit

Native TR-.beta. (T3, T4, Triac, Dimit, GC-1)

Triac>GC-1.gtoreq.T3>T4>Dimit

Variant TR-.beta. (N331S) (T3, Triac, GC-1)

Triac.apprxeq.T3>GC-1.

Scatchard Analysis of [.sup.125 I]T.sub.3 Binding to TR

Human TR-.alpha. (left panel) or human TR-.beta. (right panel) is assayed for T.sub.3 binding in the presence of increasing concentrations of [.sup.125 I]T.sub.3. The apparent equilibrium dissociation constant (20 pM for .alpha. and 67 pM for.beta.) is calculated by linear regression analysis and is depicted in FIGS. 17A-17B.

3,5-DIBROMO-4-(3',5'-DIISOPROPYL-4'-HYDROXYPHENOXY) BENZOIC ACID IS A TR-.alpha. SELECTIVE SYNTHETIC LIGAND. ##STR4##

3,5-dibromo-4-(3',5'-diisopropyl-4'-hydroxyphenoxy) benzoic acid (Compound 11), the structures of which is drawn above, is assayed for binding to the two different isoforms of the TR, TR-.alpha. and TR-.beta.. Compound 11 exhibits an IC50 of1.6 .mu.M for binding to TR-.alpha. and an IC50 of 0.91 .mu.M for binding to TR-.beta.. Assays for determining selective binding to the TR-.alpha. or TR-.beta. LBD can include reporter assays, as described herein. See also Hollenberg, et al., J.Biol. Chem., (1995) 270(24):14274-14280.

EXAMPLE 6--PREPARATION AND PURIFICATION OF A TR-.alpha. LBD

Rat TR-.alpha. LBD, residues Met122-Val410, is purified from E. coli ("LBD-122/410"). The expression vector encoding the rat TR-.alpha. LBD is freshly transfected into E. coli strain BL21(DE3) and grown at 22.degree. C. in a 50-literfermenter using 2.times. LB medium. At an A.sub.600 of 2.5-3, IPTG is added to 0.5 mM and growth is continued for 3 h before harvesting. The bacterial pellet is quickly frozen in liquid nitrogen and stored at -70.degree. C. until processed. Extraction and purification steps are carried out at 4.degree. C. The bacteria are thawed in extraction buffer (20MM Hepes, pH 8.-, 1 mM EDTA, 0.1% MTG, 0.1 mM PMSF, and 10% glycerol) at a ratio of 10 ml buffer/g bacteria. Bacteria are lysed byincubation for 15 min. with 0.2 mg/ml lysozyme and sonicated at maximum power while simultaneously homogenized with a Brinkmann homogenizer (Model PT 10/35 with generator PTA 35/2) until the solution loses its viscosity. After centrifugation for 10 minat 10,000 g, the supernatant is adjusted to 0.4 M KCl, treated with 0.6% PEI to precipitate fragmented DNA, and centrifuged for 10 min at 10,000 g. The rat TR-.alpha. LBD in the supernatant is then precipitated with 50% ammonium sulfate and centrifugedfor 10 min at 10,000 g. The precipitate is resuspended with buffer B (20 mM Hepes, pH 8.0, 1 mM EDTA, 1 mM DTT, 0.1 mM PMSF, 0.01% Lubrol, and 10% glycerol) to a final conductivity of 9 mS/cm (approx. 0.7 M ammonium sulfate) and centrifuged 1 h at100,000 g. The supernatant is frozen in liquid nitrogen and stored at -70.degree. C.

The crude extract is thawed, bound with a tracer amount of [.sup.125 I]T.sub.3, and loaded directly onto a phenyl-Toyopearl hydrophobic interaction column (2.6.times.18 cm, 95 ml bed volume) at 1.5 ml/min. The column is eluted with a 2-h gradientfrom 0.7 ammonium sulfate, no glycerol to no salt, 20% glycerol in buffer C (20 mM Hepes, pH 8.0, 0.5 mM EDTA, 1 mM DTT, 0.2 mM PMSF). The rat TR-.alpha. LBD prebound to tracer [.sup.125 I]T.sub.3 (less than 0.005% of total rat TR-.alpha. LBD) isdetected using a flow-through gamma emission detector, whereas unliganded rat TR-.alpha. LBD is assayed by postcolumn [.sup.125 I]T.sub.3 binding assays (described herein).

The phenyl-Toyopearl unliganded rat TR-.alpha. LBD peak fractions are pooled, diluted with buffer B to a conductivity of 0.5 mS/cm (equivalent to approx. 20 mM ammonium sulfate), loaded onto a TSK-DEAE anion-exchange column (2.times.15 cm, 47 mlbed volume) at 4 ml/min, and eluted with a 60-min gradient from 50 to 200 mM NaCl in buffer B.

The unliganded rat TR-.alpha. LBD peak fractions from TSK-DEAE are pooled, diluted twofold with buffer B, loaded at 0.75 ml/min on a TSK-heparin HPLC column (0.8.times.7.5 cm, 3 ml bed volume), and eluted with a 50 to 400 mM NaCl gradient inbuffer B.

The pool of unliganded rat TR-.alpha. LBD peak fractions from the TSK-heparin column is adjusted to 0.7 M ammonium sulfate, loaded at 0.75 ml/min on a TSK-phenyl HPLC column (0.8.times.7.5 cm, 3 ml bed volume), and eluted with a 60-min gradientfrom 0.7 M ammonium sulfate without glycerol to no salt with 20% glycerol in buffer C. The fractions containing unliganded rat TR-.alpha. LBD are pooled and incubated with a five fold excess of hormone for 1 h, the salt concentration is adjusted to 0.7M ammonium sulfate, and the sample is reloaded and chromatographed on the same column as described above.

EXAMPLE 7--CRYSTALLIZATION OF LIGANDED TR-.alpha. LBD

Material from a single LBD-122/410 preparation is divided into batches, and quantitatively bound with one of the following ligands: Dimit, T.sub.3, or Triac IpBr.sub.2 (3,5dibromo-3'isopropylthyronine) for the final purification step.

To maintain full saturation of rat TR-.alpha. LBD with a ligand, and to prepare the complex for crystallization, the ligand-bound rat TR-.alpha. LBD is concentrated and desalted in an Amicon Centricon-10 microconcentrator (McGrath et al,Biotechniques, (1989) 7:246-247, incorporated by reference herein), using 10 mM Hepes (pH 7.0), 3.0 mM DTT, and 1.0 nM to 10 nM ligand.

Factorial crystallization screening trials (Jancarik & Kim, J. Appl. Crystallogr. (1991) 24:409-411, incorporated by reference herein) are carried out for rat TR-.alpha. LBD bound to selected ligands using hanging-drop vapor diffusion at17.degree. C. (with 1 .mu.l protein solution, 1 .mu.l precipitant solution and a 0.5 ml reservoir using silanized coverslip: (McPherson, Preparation and Analysis of Protein Crystals (1982), incorporated by reference herein). Rat TR-.alpha. LBD is notstable at 4.degree. C. and is stored at -80.degree. C., where it maintains its avidity for hormone and its crystallizability for approximately two to three months. These procedures are carried out as described in McGrath, M. E. et al., J. Mol. Biol. (1994) 237:236-239 (incorporated by reference,. Crystals are obtained in condition 21 of the screening trials (Jancarik & Kim 1991) and conditions are then optimized. Wedge-shaped crystals are reproducibly obtained with hanging-drop vapor fusion at22.degree. C. with 15% 2-methyl-2,4-pentanediol (MPD), 0.2 M ammonium acetate and 0.1 M sodium cacodylate (pH 6.7), 3 mM DTT, with 2 .mu.l protein solution, 1 .mu.l precipitant solution and a 0.6 ml reservoir using silanized coverslip, and with 8.7mg/ml (Dimit), 5.5 mg/ml (IpBr.sub.2), 5 mg/ml (Triac), or 2.3 mg/ml (T.sub.3) over a period of three days. Under these conditions, diffraction quality crystals (dimension 0.5.times.0.2.times.0.0075 mm.sup.3) can be grown at ambient temperature(22.degree. C.). The best crystals have a limiting dimension of approximately 100 .mu.m and are obtained at a protein concentration between 2.3 and 8.7 mg/ml in the presence of 3 mM DTT. The crystals are of the monoclinic space group C2, with onemonomer in the asymmetric unit.

EXAMPLE 8--CRYSTALLIZATION OF HUMAN TR-.beta. LBD COMPLEXED WITH T3, TRIAC, OR GC-1

Human TR-.beta. LBD complexed with T.sub.3, Triac, or GC-1 are purified according to the same procedures described above for the rat TR-.alpha. LBD, with the following modifications.

The expression of human TR-.beta. LBD differs from the rat TR-.alpha. LBD in that the human TR-.beta. LBD residues extend from the amino acid at position 716 through the amino acid at position 1022, according to the amino acid numbering schemefor the various nuclear receptor LBDs depicted in illustrates a numbering scheme applicable to all of the nuclear receptors listed as well as to any additional homologous nuclear receptors. The vertical lines on FIGS. 3A-3R at position 725 and atposition 1025 delineate the preferred minimum amino acid sequence necessary to obtain adequate binding of ligand. The amino acid sequence from position 716 to position 1022 according to the numbering scheme of FIGS. 3A-3R corresponds to the amino acidpositions 202 to 461 according to the conventional numbering of the amino acid sequence of human TR-.beta. which is publicly available. Also, the human TR-.beta. LBD is expressed with a histidine tag, as described in Crowe et al., Methods in MolecularBiology (1994) 31:371-387, incorporated by reference herein.

The purification of human TR-.beta. LBD is the same as that described above for the rat TR-.alpha. LBD with the following exceptions. First, before the purification step using the hydrophobic interaction column, a step is added in which theexpressed human TR-.beta. LBD is purified using a nickel NTA column (commercially available from Qiagen, Chatsworth, Calif.) according to manufacturer's instructions, and eluted with 200 mM imidazole. The second difference is that in the purificationof the human TR-.beta. LBD, the purification step using a heparin column is omitted.

The crystallization of human TR-.beta. LBD bound to T.sub.3, Triac or GC-1 is as follows. Crystals are obtained in condition 7 of the factorial screen using hanging drops as before at ambient temperature (22.degree. C.) using the factorialcrystallization screening trials of Jancarik & Kim (1991) and using the commercially available product from Hampton Research, Riverside). The following are optimum conditions: hexagonal bipyrimidal crystals are grown at 4.degree. C. for 2-3 days fromhanging drops containing 1.0-1.2 M sodium acetate (pH unadjusted) and 0.1 M sodium cacodylate (pH 7.4), 3 mM DTT, with either a 1 .mu.l protein solution, 1 .mu.l precipitant solution or 2 .mu.l protein solution, 1 .mu.l precipitant solution and a 0.6 mlreservoir using silanized coverslip, at a protein concentration of 7-10 mg/ml. The best crystals have a limiting dimension of 200 .mu.m. The following are optimum conditions for crystallization of the TR-.beta. LBD with GC-1: hexagonal bipyrimidalcrystals are grown at 4.degree. C. for 2-3 days from hanging drops containing 0.8-1.0M sodium acetate (pH unadjusted), 50-200 nM sodium succinate, and 0.1M sodium cacodylate (pH 7.2), 3 mM DTT, 1 .mu.l protein solution, 1 .mu.l precipitant solution anda 0.6 ml reservoir using silanized coverslip, at a protein concentration of 7-10 mg/ml. The best crystals have a limiting dimension of 200 .mu.M. The unit cell dimensions are cell length a=b=68.73, cell length c=130.09. The unit cell angles are.alpha.=90.degree., .beta.=90.degree., .gamma.=120.degree..

The crystal system for human TR-.beta. LBD bound to T.sub.3, Triac or GC-1 is trigonal with the space group p3.sub.1 21. The unit cell dimensions are cell length a=cell length b=68.448 angstroms, cell length c=130.559 angstroms. The angles are.alpha.=90.degree., .beta.=90.degree., gamma=120.degree..

EXAMPLE 9--DETERMINATION OF LIGANDED TR-.alpha. LBD AND TR-.beta. CRYSTAL STRUCTURES

Data from each cocrystal (Rat TR-.alpha. LBD with Dimit, T3 and IpBr2; Human TR-.beta. LBD with Triac and GC-1) is measured on a Mar area detector at Stanford Synchrotron Radiation Laboratory beamline 7-1 (.lambda.=1.08 angstroms) using1.2.degree. oscillations. Data from the cocrystal of the hTR-.beta. LBD with Triac is measured on a Mar area detector at Stanford Synchrotron Radiations Laboratory beamline 7-1 (.lambda.=1.08 angstroms) using 1.0 oscillations. Data from the cocrystalof the hTR-.beta. LBd with GC-1 is measured on a R-axis II area detector on a Rigaku rotating Cu anode (50 kV, 300 mA). The crystals are transferred into a cryosolvent containing 1.2M sodium acetate, 0.1M sodium cacodylate, and 15% glycerol followed bya second transfer into 30% glycerol, then flash frozen in liquid nitrogen. An orientation matrix for each crystal is obtained using DENZO. The reflections are integrated with DENZO (commercially available from Molecular Structure Corp., The Woodlands.,Tex.) and are scaled with SCALEPACK (as described in Otwinowski, Z, Proceedings of the CCP4 Study Weekend: "Data Collection and Processing," 56-62 (SERC Daresbury Laboratory, Warrington, UK 1993) incorporated by reference).

For rTR-.alpha. cocrystals, data from the T.sub.3 cocrystal is measured with the b* axis approximately parallel with the spindle. The crystals are flash frozen at -178.degree. C. in a nitrogen gas stream with the MPD mother liquor serving asthe cryosolvent. An orientation matrix for each crystal is determined using REFIX (Kabsch, W., J. Appl. Crystallogr. (1993) 26:795-800 incorporated by reference). Reflections are integrated with DENZO, and are scaled with SCALEPACK.

For the T.sub.3 data set, Bijvoet pairs are kept separate, and are locally scaled using MADSYS (W. Hendrickson (Columbia University) and W. Weis (Stanford University)).

Cocrystals prepared from the three isosteric ligands are isomorphous. MIR analysis is performed using programs from the CCP4 suite (Collaborative Computational Project, N.R. Acta Crystallogr. (1994) D50:760-763, incorporated by referenceherein). Difference Pattersons is calculated for both T.sub.3 and IpBr.sub.2, taking the Dimit cocrystal as the parent. The positions of the three iodine atoms in the T.sub.3 difference Patterson are unambiguously determined from the Harker section ofthe density map as peaks of 11.sigma. above background. The positions for the two bromine atoms in the IpBr.sub.2 cocrystals, are located independently, as peaks 8.sigma. above the noise level. Phases for the LBD-122/410 are calculated from thesolution to the IpBr.sub.2 difference Patterson, and are used to confirm the location of the unique third iodine of the T.sub.3 cocrystal. Halogen positions are refined with MLPHARE, including the anomalous contributions from the iodine atoms(Otwinowski, Z, Proceedings of the CCPR Study Weekend 80-86 (SERC Daresbury Laboratory, Warrington, UK 1991)). The MIRAS phases are improved through solvent flattening/histogram matching using DM (Cowtan, K., Joint CCP4 and ESF-EACBM Newsletter onProtein Crystallography (1994) 31: 34-38, incorporated by reference herein).

A model of the LBD-122/410 with Dimit bound is built with the program O from the solvent flattened MIRAS 2.5 angstrom electron density map (Jones et al., Acta Crystallogr. (1991) A 47:110-119, incorporated by reference herein). The initialmodel, without ligand, (Rcryst=40.1%), is refined using least-squares protocols with XPLOR. The Dimit ligand is built into unambiguous Fo-Fc difference density during the following round. Subsequent refinement employs both least-squares and simulatedannealing protocols with XPLOR (Brunger et al., Science (1987) 235:458-460), incorporated by reference herein). Individual atomic B-factors are refined isotropically. As defined in PROCHECK, all residues are in allowed main-chain torsion angle regionsas described in Laskowski et al., J. Appl. Crystallogr., (1993) 26:283-291, incorporated by reference herein. The current model is missing 34 residues (Met.sub.122 -Gln.sub.156) at the N-terminus, and 5 residues (Glu.sub.406 -Val.sub.410) at theC-terminus.

In addition, the following residues are not modeled beyond C.beta. due to poor density: 184, 186, 190, 198, 206, 209, 240, 301, 330, 337, 340, 343, 359, and 395. The average B-value for protein atoms is 34.5 .ANG..sup.2. The final modelconsists of the LBD-122/410, residues Arg.sub.157 -Ser.sub.183, Trp.sub.185 -Gly.sub.197, Ser.sub.199 -Asp.sub.206 and Asp.sub.208 -Phe.sub.405 ; three cacodylate-modified cysteines: Cys.sub.334, Cys.sub.380 and Cys.sub.392 ; and 73 solvent moleculesmodeled as water (2003 atoms).

The occupancy for the two bromine sites is set to 35 electrons. The occupancies of the iodine sites are relative to this value. .sctn.Phasing power=<FH>/<.epsilon.>, where <FH> is the mean calculated heavy atom structurefactor amplitude and <.epsilon.> is the mean estimated lack of closure. .parallel.Rcullis=<.epsilon.>/<iso>, where <.epsilon.> is the mean estimated lack of closure and (iso) is the isomorphous difference. .paragraph.Rcryst=100.times..SIGMA..sub.hkl.vertline.F.sub.o -Fc.vertline./.SIGMA..sub.hkl.vertline.F.sub.o.vertline. where F.sub.o and F.sub.c are the observed and calculated structure factor amplitudes (for data F/.sigma.>2). The Rfree wascalculated using 3% of the data, chosen randomly, and omitted from the refinement. .sctn. Correlation coefficient=.SIGMA..sub.hkl (.vertline.F.sub.o.vertline.-.vertline.F.sub. o.vertline.).times.(.vertline.F.sub.c.vertline.-.vertline.F.sub. c.vertline.)/.SIGMA..sub.hkl (.vertline.F.sub.o.vertline.-.vertline.F.sub.o.vertline.).sup. 2.times..SIGMA..sub.hkl (.vertline.F.sub.c.vertline.-.vertline.F.sub.c.vertline.).sup.2

EXAMPLE 10 PHASING OF THE rTR-.alpha. LBD AND hTR-.beta. LBD COMPLEX WITH TRIAC

Due to the possible non-isomorphism of the rTR.alpha. LBD complex with Triac, a molecular replacement solution is determined using AMORE (Navaza, J., Acta Crystallographica Section A-Fundamentals of Crystallography (1994) 50:157-63 from astarting model consisting of rTR.alpha. LBD complex with T.sub.3, but with the ligand, all water molecules, and the following residues omitted: Asn 179, Arg228, Arg262, Arg266, and Ser 277. Strong peaks are obtained in both the rotation and translationsearches, with no significant (>0.5 times the top peak) false solutions observed (Table 6). Strong positive density present in both the anomalous and conventional difference Fourier maps confirm the solution. Maps are calculated using sigma-Aweighted coefficients output by REFMAC (Murshudov, et al. "Application of Maximum Likelihood Refinements," in Refinement of Protein Structures, Proceedings of Daresbury Study Weekend (1996)) after 15 cycles of maximum likelihood refinement. Triac, theomitted residues, and water molecules 503, 504, 534 (following the numbering convention for the TR complex with T3) are built into the resulting difference density using O (Jones et. al.); the conformations of these residues are further confirmed in asimulated-annealing omit map (Brunger et. al.). The complete model is then refined using positional least-squares, simulated annealing, and restrained, grouped B factor refinement in XPLOR to an Rcryst of 23.6% and an Rfree of 24.1%

Phasing of a related LBD using the structure of the rTR-.alpha. LBD is conducted as follows. A molecular replacement solution for the hTR-.beta. LBD complex with Triac is determined using AMORE from a starting model consisting of therTR-.alpha. LBD complexed with T3, but with the ligand and all water molecules omitted. Strong peaks are obtained in both the rotation and translation searches, with no significant (>0.5 times the top peak) false solutions (Table 7). Strongpositive density present in both the anomalous and conventional difference Fourier maps confirm the solution. Initial maps are calculated using sigma-A weighted (coefficients output by REFMAC after 9 cycles of maximum likelihood refinement. Thereal-space fit for each residues was calculated using OOPS (Kleywegt, GJ and Jones, TA, OOPS-a-daisy, ESF/CCP4 Newsletter Jun. 30, 1994, pp. 20-24) and the residues with a real-space fit less than 2 standard deviations below the mean removed:Ala253-Lys263; Glu245-Leu250. To reduce bias, the following residues were modeled as alanine: Arg282, Arg316, Arg 320, Asn 331. Cycles of rebuilding and positional least-squares, simulated annealing, and restrained, grouped B factor refinement withXPLOR produce a model with an R.sub.cryst of 25.3 and an R.sub.free of 28.9%. The final model consists of hTR-.beta. LBD residues Glu202-Gln252, Val264-Glu460; three cacodylate-modified cysteines with the cacodylate moeity modeled as free arsenic:Cys294, Cys298, Cys388, and Cys434; and 35 solvent molecules in modeled as water.

EXAMPLE 11 CONNECTING QSAR WITH STRUCTURE IN THE THYROID HORMONE RECEPTOR

The conclusions of classic thyroid hormone receptor quantitative structure-activity relationships may be summarized as follows:

1) the R.sub.4 '-hydroxyl group functions as a hydrogen bond donor;

2) the amino-propionic acid interacts electrostatically through the carboxylate anion with a positively charged residue from the receptor;

3) the preferences of R.sub.3 /R.sub.5, substituent are I>Br>Me>>H;

4) the preferences of the R.sub.3 '-substituent are Ipr>I>Br>Me>>H.

The structure of the thyroid hormone receptor ligand binding domain complexed with the agonists T3, IpBr.sub.2, Dimit, Triac, and GC1 as provided herein, permits:

1) the identification of receptor determinants of binding at the level of the hydrogen bond;

2) the association of these determinants with the predictions of classic thyroid hormone receptor QSAR; and

3) prediction as to which determinants of binding are rigid, and which are flexible, for both the ligand and the receptor.

This classification for the agonists of the type (R.sub.1 =amino-propionic, acetic acid; R.sub.3,R.sub.5 =I,Br,Me; R.sub.3 '=Ipr,I) is given below (for the representative ligand T.sub.3);

F=Fiducial (always satisfied)

A=Adjustable ##STR5##

Based upon the methods and data described herein, the following is an embodiment of the computational methods of the invention, which permit design of nuclear receptor ligands based upon interactions between the structure of the amino acidresidues of the receptor LBD and the four different ligands described herein. The small molecule structures for the ligands can be obtained from Cambridge Structural Database (CSD), and three dimensional models can be constructed using the methodsdescribed throughout the specification. The following are factors to consider in designing synthetic ligands:

1) Histidine 381 acts as a hydrogen bond acceptor for the R.sub.4 ' hydroxyl, with the optimal taulomer maintained by water molecules. See FIG. 23 and FIG. 24. Histidine is the only hydrophilic residue in this hydrophobic pocket that surroundsthe R.sub.4 ' substituent. Histidine can be either a hydrogen bond acceptor or donor, depending on its tautomeric state. It is preferably a hydrogen bond donor, but can tolerate being a hydrogen bond acceptor, as for example, when there is a methoxy atthe R.sub.4 ' position of the ligand;

2) Arginines 228, 262, and 266 interact directly and through water-mediated hydrogen bonds with the R.sub.1 -substituent, with the electrostatic interaction provided by Arginine 266 (as in the Triac complex). This polar pocket is illustrated byFIG. 23-FIG. 25. FIG. 23 depicts T.sub.3 in the TR.alpha. ligand binding cavity, where T3's amino-propionic R1-substituent interacts with Arg 228, HOH502, H9H503 and HOH504 via hydrogen bonds. FIG. 24 depicts Triac in the ligand binding cavity, withits --COOH R.sub.1 substituent in the polar pocket. In FIG. 24, Arg 228 no longer shares a hydrogen bond with the ligand, but the --COOH R.sub.1 substituent forms hydrogen bonds with Arg 266. FIG. 25 superimposes T.sub.3 and Triac in the ligand bindingcavity and shows several positionally unchanged amino acids and water molecules, and selected changed interacting amino acids and water molecules. The three figures illustrate parts of the polar pocket that can change and those parts that do not moveupon binding of different ligands. For example, the Arg 262 at the top of the polar pocket does not move, even when the R.sub.1 substituent has changed from a --COOH to an aminopropionic acid group. However, the other two Arginines, Arg 228 and Arg266, demonstrate flexibility in the polar pocket to respond to the change in the size or chemical nature of the R.sub.1 substituent.

3) Inner and outer pockets for the R.sub.3 /R.sub.5 substituents are formed by Ser260, Ala263, Ile299; and Phe 218, Ile221, Ile222, respectively. See FIGS. 21 and 22. The inner pocket is filled by either the R.sub.3 or the R.sub.5 substituent,regardless of the size of the substituent, and may act as a binding determinant by positioning the ligand in the receptor. Optimally, the inner pocket amino acids interact with an R3 or R5 substituent that is no larger than an iodo group. If the innerpocket is filled by the R.sub.3 substituent, then the outer pocket interacts with the R.sub.5 substituent and vice versa. The outer pocket can adjust to the size of its substituent through main chain motion centered at the break in helix 3(Lys220-Ile221), suggesting that the bending of H3, and motion of the N-terminal portion of H3, may represent a conformational change induced on ligand binding. The outer pocket has greater flexibility than does the inner pocket in terms ofaccommodating a larger substituent group.

4) A pocket for the R.sub.3 '-substituent is formed by Phe 215, Gly290, Met388. The pocket is incompletely filled by the R.sub.3 '-iodo substituent, and accommodates the slightly larger 3'-isopropyl substituent by movement of the flexible Met388side chain and the H7/H8 loop. This pocket can accommodate R.sub.3 ' substituents that are even larger than isopropyl, for example, a phenyl group.

The above information will facilitate the design of high affinity agonists and antagonists by improving automated QSAR methodologies and informing manual modeling of pharmaceutical lead compounds. For example, the inclusion of discrete watermolecules provides a complete description of hydrogen bonding in the polar pocket for use with pharmacophore development: also, the identification of mobile and immobile residues within the receptor suggests physically reasonable constraints for use inmolecular mechanics/dynamics calculations.

EXAMPLE 12 DESIGN OF AN INCREASED AFFINITY LIGAND

Direct interaction between the receptor and the ligand is limited in the polar pocket, which interacts with the R.sub.1 substituent. While the lack of complementarity may contain implications for biological regulation, it also provides anopportunity for increasing affinity by optimizing the interaction between the amino acids of the polar pocket and the R.sub.1 substituent of a synthetic ligand. The structure of the receptor-ligand interactions described herein enables design of anincreased affinity synthetic ligand having two complementary modifications:

1) Remove the positively charged amine. The strongly positive electrostatic potential predicted for the polar pocket suggests that the positively charged amine of the aminopropionic acid R.sub.1 substituent may be detrimental to binding. Suitable groups for substitution are suggested by the nature of nearby hydrogen bond partners: for example, Thr 275 O or Ser 277 N. See e.g. Tables in Appendix 2. For example, any any negatively charged substituent would be compatible for interactingwith the amino acids of the polar pocket, including carboxylates, carbonyl, phosphonates, and sulfates, comprising 0 to 4 carbons. Another example of an R.sub.1 substitution is an oxamic acid that replaces the amine of the naturally occurring ligandwith one or more carbonyl groups.

2) Incorporate hydrogen bond acceptor and donor groups into the R.sub.1 -substituent to provide broader interactions with the polar pocket scaffold. Such hydrogen bond acceptor and donor groups incorporated into the R.sub.1 -substituent willallow interactions that would otherwise occur with water molecules in the polar pocket. Specific waters include HOH 504 (hydrogen bonds with Ala 225 O and Arg 262 NH); and HOH 503 hydrogen bonds with Asn 179 OD1, Ala 180 N), both of which are present inall four complexes (TR LBD complexed with T3, TR LBD complexed with IpBr.sub.2, TR LBD complexed with Dimit and TR LBD complexed with Triac). Analysis of the hydrogen bonding network in the polar pocket suggests replacement of HOH 504 with a hydrogenbond acceptor, and HOH 503 with an hydrogen bond donor (although the chemical nature of asparagine probably permits flexibility at this site). Thus, incorporating a hydrogen bond acceptor in an R1 substituent that could take the place of the HOH504 orincorporating a hydrogen bond acceptor in an R1 substituent that could positionally replace the HOH503, or a combination thereof, are methods of designing novel synthetic TR ligands.

These two design approaches can be used separately or in combination to design synthetic ligands, including those in Table 5 (below).

A corollary to this approach is to design specific interactions to the residues Arg262 and Asn 179. The goal is to build in interactions to these residues by designing ligands that have R.sub.1 substituents that form hydrogen bonds with watermolecules or charged residues in the polar pocket.

High-affinity ligands also may be designed and selected using small molecules that bind to proximal subsites of the target nuclear hormone receptor that are identified in a structure-based screen and then linked together in their experimentallydetermined bound orientiations. Such a method has been described in design of high-affinity ligands for the FK506 binding protein (FKBP), stromelysin, gelatinase A, and human papillomavirus E2 (Hajduk et al., Science 278:497-499 (1997)), which referenceand its references are incorporated herein by reference. The preferred small molecules for screening are compounds of Formula I or derivatives thereof. For example, a compound of Formula I (.phi.-X-.phi.) or a derivative thereof (.phi.-X or X-.phi.) isscreened for binding a target nuclear hormone receptor LBD. Proximal subsites of the nuclear hormone receptor include the hydrophobic and polar pockets of the LBD, and subsites extended therefrom. As an example, Fourier transformation or nuclearmagnetic resonance (NMR)-based structure screens can be used. When a NMR-based screen is used, binding can be detected from the amide chemical shift changes observed in two-dimensional heteronuclear single quantum correlation (HSQC) spectra acquired inthe presence and absence of added compound. Once two ligands are identified that bind to the receptor, the crystal or solution structure of the ternary complex is determined. From the structural information, a compound is synthesized which links thetwo ligands, where the linker is selected based on structural information. The new compound is then screened for binding affinity, for example, using a binding assay as described herein. Only a few linked ligands need to synthesized and screened whenusing this approach.

Compounds of the invention also may be interactively designed from structural information of the compounds described above using other structure-based design/modeling techniques (Jackson, R. C., Contributions of protein structure-based drugdesign to cancer chemotherapy. Seminars in Oncology, 1997, 24(2)L164-172; and Jones, T. R., et al., J. Med. Chem., 1996 39(4):904-917).

TABLE 5 Synthetic TR Ligands ##STR6## R1 R2 R3 R5 R6 X R'2 R'3 R'4 R'5 R'6 CO.sub.2 H H Me Me H O H Me OH Me H CH.sub.2 CO.sub.2 H I I S Et SH Et CH.sub.2 CH.sub.2 CO.sub.2 H Br Br nPr NH.sub.2 nPr CH.sub.2 CH(NH.sub.2)CO.sub.2 H Cl CliPr iPr OCH.sub.2 CO.sub.2 H Et Et Ph nBu OCH.sub.2 CH.sub.2 CO.sub.2 H OH OH I nPen NHCH.sub.2 CO.sub.2 H NH.sub.2 NH.sub.2 Br nHex NHCH.sub.2 CH.sub.2 CO.sub.2 H SH SH Cl Ph CH.sub.2 COCOCO.sub.2 H hetero cycle NHCOCOCO.sub.2 H aryl COCO.sub.2 H CF.sub.2 CO.sub.2 H COCH.sub.2 CO.sub.2 H

Any combination of the above substituents in the biphenyl ether scaffold structure shown above may result in a potentially pharmacologically useful ligand for the thyroid hormone receptor. These novel ligands may be antagonists of the thyroidreceptor.

TABLE 6 TR-.alpha. LBD-122/410 Dimit T3 IpBr.sub.2 Triac Data collection Cell dimensions a (.ANG.) 117.16 117.19 117.18 118.19 b (.ANG.) 80.52 80.20 80.12 81.37 c (.ANG.) 63.21 63.23 63.13 63.73 .beta. (.degree.) 120.58 120.60 120.69121.00 Resolution (.ANG.) 2.2 2.0 2.1 2.45 Obs. Reflections, (no.) 57031 64424 66877 83573 Unique Reflections, 22327 21023 23966 18453 (no.) Completeness, (%) 87.0 82.4 93.7 96.0 *R.sub.sym (%) 3.9 3.5 4.5 7.5 Phasing (15.0-2.5 .ANG.) .dagger.R.sub.der (%) -- 19.6 11.6 No. of sites -- 3 2 .dagger-dbl.Occupancy -- 44.6 (19.8) 35.0 (Anomalous) -- 50.2 (23.7) 35.0 39.2 (22.3) .sctn.F.sub.H /E centric (acentric) 15.0-5.0 .ANG. -- 3.67 (4.61) 2.25 (3.09) 5.0-3.0 .ANG. -- 2.23(2.75) 1.25 (1.85) 3.0-2.5 .ANG. -- 1.64 (1.99) 1.15 (1.57) .parallel.R.sub.cullis (%) 15.0-5.0 .ANG. -- 33 44 5.0-3.0 .ANG. -- 45 63 3.0-2.5 .ANG. -- 60 65 Mean figure of merit 0.62 -- -- MR Phasing (10-3.5 .ANG.) Rotation Search: .theta..sub.1= 309.37 Euler Angles (.degree.) .theta..sub.2 = 48.96 .theta..sub.3 = 127.28 .sctn. correlation 34.3 coefficient Translation Search: x = 0.1571 Fractional coordinates y = 0.000 z = 0.3421 .sctn. correlation 65.8 Coefficient .sup.t R factor31.2 Refinement 15.0-2.2 5.0-2.0 15.0-2.2 25-2.5 Resolution (.ANG.) .paragraph.R.sub.cryst (%) 20.5 22.1 21.4 23.6 R.sub.free (%) 22.7 24.0 22.4 24.1

TABLE 7 TR-.beta. LBD-202/461 Triac T3 GC1 Data collection Space Group P3121 Cell dimensions a (.ANG.) 68.9 68.45 68.73 c (.ANG.) 131.5 130.56 130.09 Resolution (.ANG.) 2.4 3.1 2.8 Obs. Reflections, (no.) 80196 55103 54104 UniqueReflections, (no.) 14277 6847 8987 Coverage (%) 97.0 95.7 97.1 *R.sub.sym (%) 5.1 4.6 5.5 MR Phasing (15.0-3.5 .ANG.) Rotation Search .theta..sub.1 = 39.13 Euler Angles (.degree.) .theta..sub.2 = 68.00 .theta..sub.3 = 323.6 .sctn. correlationcoefficient 21.6 (Highest false peak) (10.8) Translation Search x = 0.748 Fractional Coordinates y = 0.158 z = 0.167 .sctn. correlation coefficient 57.5 (Highest false peak) (38.7) 0.612 *R factor 40.7 40.8 Refinement Resolution (.ANG.) 30-2.430-2.9 .paragraph.R.sub.cryst (%) 25.3 27.3 R.sub.free (%) 28.9 33.4

All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to beincorporated by reference. The nuclear receptor ligands, particularly the TR ligands, of these references are herein incorporated by reference and can be optionally excluded from the claimed compounds with a proviso.

Headings and subheadings are presented only for the convenience of the reader and should not be used to construe the meaning of terms used within such headings and subheadings.

The invention now being fully described, it will be apparent to one of ordinary skill in the art that many changes and modifications can be made thereto without departing from the spirit or scope of the appended claims.

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TABLE 8 APPENDIX 2 Dimit Amino Acid Amino Acid Distance Atom in full length .alpha. Atom A C16 215-PHE CD1 3.98 C16 215-PHE CE1 3.86 C19 218-PHE O 3.69 C16 218-PHE CB 3.89 C18 218-PHE CB 3.92 C19 218-PHE CB 4.13 C18 218-PHE CD2 3.77 C16 219-THR CG2 3.68 C19 221-ILE CG1 4.11 C6 222-ILE CD1 4.18 C8 222-ILE CD1 3.72 C10 222-ILE CD1 3.53 C12 222-ILE CD1 3.85 O1 222-ILE CD1 4.13 C13 225-ALA C8 3.64 O4 225-ALA C8 4.02 O4 228-ARG CZ 3.96 C17 228-ARG NH2 3.36 O3 228-ARG NH2 3.58 O4 228-ARG NH2 2.86 C10 256-MET SD 3.70 C12 256-MET SD 3.89 C10 256-MET CE 3.88 C12 256-MET CE 3.83 C11 259-MET C 4.03 C11 259-MET O 3.66 C15 259-MET O 3.42 N1 259-MET O 3.71 C1 259-MET C8 4.20 C11 259-MET C8 3.87 C13 259-MET C8 4.09 C15262-ARG C8 4.03 C17 262-ARG C8 3.58 O3 262-ARG C8 3.62 O4 262-ARG C8 3.85 C17 262-ARG CD 4.10 O4 262-ARG CD 3.61 N1 263-ALA N 3.71 C17 263-ALA CA 3.69 N1 263-ALA CB 3.46 O3 266-ARG NH1 3.93 N1 275-THR O 3.62 N1 276-LEU CA 3.51 N1 276-LEU C3.92 C5 276-LEU CD1 4.05 C19 276-LEU CD1 4.04 C7 276-LEU CD2 4.09 C9 276-LEU CD2 3.95 C11 276-LEU CD2 4.13 N1 276-LEU CD2 4.17 C13 277-SER N 4.14 C15 277-SER N 3.79 Dimit Amino Acid Distance Atom in full length .alpha. Atom A C17 277-SER N3.69 N1 277-SER N 3.30 O3 277-SER N 3.19 C17 277-SER CA 3.92 O3 277-SER CA 3.35 C13 277-SER OG 3.92 C7 287-LEU CD2 3.90 C18 290-GLY C 4.04 C18 290-GLY O 3.54 C18 291-GLY CA 4.04 C18 292-LEU N 4.20 C2 292-LEU CG 4.18 C4 292-LEU CG 3.86 C6292-LEU CG 4.01 C2 292-LEU CD1 3.88 C4 292-LEU CD1 4.02 O2 292-LEU CD1 4.07 C4 292-LEU CD2 4.05 C6 292-LEU CD2 3.72 C8 292-LEU CD2 3.69 C10 292-LEU CD2 3.98 O1 292-LEU CD2 4.16 C20 299-ILE CD1 3.87 C8 381-HIS CD2 3.90 C10 381-HIS CD2 3.84 O1381-HIS GO2 3.40 O1 381-HIS CE1 3.72 C8 381-HIS NE2 3.47 C10 381-HIS NE2 3.51 O1 381-HIS NE2 2.64 C6 388-MET CE 3.90 C8 401-PHE CE1 4.19 O1 401-PHE CE1 3.37 C16 401-PHE CZ 3.97 O1 401-PHE CZ 3.28 N1 502-H.sub.2 O O1 3.35 O3 502-H.sub.2 O O12.56 O3 503-H.sub.2 O O1 3.13 O4 503-H.sub.2 O O1 3.72 O4 504-H.sub.2 O O1 2.72 Legend to Table 8. The table lists the interactions with Dimit (DMT). The column headings are as follows: #1 The atom of Dimit that interacts with the amino acid ofthe receptor. These are also numbered in FIG. 32. #2 The amino acid in the full length rTR.alpha. that interacts with the ligand. #3 The name of the atom in the amino acid (standard nomenclature) where the interaction occurs. #4 The distance in Abetween Dimit and the protein atom.

TABLE 9 Triac Amino Acid Amino Acid Distance Atom in full length .alpha. Atom A I1 218-PHE O 3.52 I1 221-ILE CD1 4.16 I1 221-ILE CG1 3.92 I1 222-ILE CA 4.15 I1 222-ILE CB 4.03 I1 222-ILE CG1 3.92 C8 222-ILE CD1 4.12 C10 222-ILE CD13.77 C12 222-ILE CD1 3.79 C13 225-ALA CB 4.17 C3 225-ALA CB 3.86 C10 256-MET SD 3.45 C12 256-MET SD 3.73 C10 256-MET CE 3.66 C12 256-MET CE 3.77 I3 256-MET CE 3.89 C1 259-MET O 3.93 C11 259-MET O 3.24 O3 259-MET O 4.09 C1 259-MET CB 3.89 C13259-MET O 3.74 C14 259-MET O 3.96 C1 259-MET CB 3.89 C11 259-MET CB 3.68 C13 259-MET CB 4.01 C11 259-MET CA 4.13 C13 259-MET CA 4.20 I3 260-SER CA 4.10 I3 260-SER OG 4.19 C14 262-ARG CB 4.07 O4 262-ARG CB 3.60 O3 263-ALA N 3.79 C14 263-ALA N4.12 O3 263-ALA CA 3.67 O3 263-ALA CB 3.49 C11 263-ALA CB 4.00 C14 266-ARG CZ 3.89 O3 266-ARG CZ 4.01 O4 266-ARG CZ 3.03 C14 266-ARG NH1 3.25 O3 266-ARG NH1 3.00 O4 266-ARG NH1 2.82 C14 266-ARG NH2 3.48 O3 266-ARG NH2 4.01 O4 266-ARG NH2 2.34 O3 275-THR C 4.02 C14 275-THR O 4.20 O3 275-THR O 3.20 O3 278-LEU CA 3.11 O3 276-LEU C 3.52 O3 276-LEU N 4.04 C14 276-LEU CA 3.98 O3 276-LEU CA 3.11 C14 276-LEU C 3.98 O3 276-LEU CB 3.95 O2 276-LEU CD1 4.03 I1 276-LEU CD1 4.10 C7 276-LEU CD23.84 C9 276-LEU CD2 3.73 CII 276-LEU CD2 4.06 O2 276-LEU CD2 4.10 O3 276-LEU CD2 3.91 C13 277-SER N 4.06 C14 277-SER N 3.13 O4 277-SER N 3.28 O3 277-SER N 3.05 C14 277-SER CA 3.76 O4 277-SER CA 3.52 C3 277-SER OG 3.87 C13 277-SER OG 4.02 C14277-SER OG 4.14 I2 290-GLY O 3.57 I2 292-LEU CG 3.94 C4 292-LEU CG 3.95 C6 292-LEU CG 3.65 C8 292-LEU CG 4.02 C2 292-LEU CD1 4.11 C4 292-LEU CD1 3.85 C6 292-LEU CD1 4.02 I2 292-LEU CD2 3.98 C4 292-LEU CD2 4.11 C6 292-LEU CD2 3.44 C8 292-LEUCD2 3.28 C10 292-LEU CD2 3.88 O1 292-LEU CD2 3.35 I3 299-ILE CD1 3.77 C8 381-HIS CD2 3.87 C10 381-HIS CD2 3.90 O1 381-HIS GO2 3.20 O1 381-HIS CE1 3.82 C8 381-HIS NE2 3.57 C10 381-HIS NE2 3.52 O1 381-HIS NE2 2.64 O1 388-MET CE 4.03 O1 401-PHECE1 3.86 O1 401-PHE CZ 3.70 C13 460-H.sub.2 O O1 4.00 Legend to Table 9. The table lists the interactions with Triac. The column headings are as follows: #1 The atom of Triac that interacts with the amino acid of the receptor. These are alsonumbered in FIG. 32. #2 The amino acid in the full length rTR.alpha. that interacts with the ligand. #3 The name of the atom in the amino acid (standard nomenclature) where the interaction occurs. #4 The distance in A between Triac and the proteinatom.

TABLE 10 IpBR.sub.2 Amino Acid Amino Acid Distance Atom in full length .alpha. Atom A C16 215-PHE CD1 4.01 C16 215-PHE CE1 3.78 BR1 218-PHE O 3.24 BR1 218-PHE C 3.98 C16 218-PHE CB 3.81 C18 218-PHE CB 3.92 BR1 218-PHE CB 4.08 C18218-PHE CD2 3.92 C16 219-THR CG2 3.45 BR1 221-ILE CG1 3.81 BR1 221-ILE CD1 4.07 BR1 222-ILE CB 3.81 BR1 222-ILE CG1 3.97 C6 222-ILE CD1 4.07 C8 222-ILE CD1 3.64 C10 222-ILE CD1 3.50 C12 222-ILE CD1 3.82 O1 222-ILE CD1 4.08 C13 225-ALA CB 3.76 O4 225-ALA CB 4.01 O4 228-ARG CZ 3.92 C17 228-ARG NH2 3.26 O3 228-ARG NH2 3.43 O4 228-ARG NH2 2.79 C10 256-MET SD 3.65 C12 256-MET SD 3.71 C10 256-MET CE 3.90 C12 256-MET CE 3.75 BR2 256-MET CE 4.03 C11 259-MET C 3.98 C11 259-MET O 3.52 C15259-MET O 3.44 N1 259-MET O 3.76 C11 259-MET CB 3.87 N1 262-ARG C 4.03 C15 262-ARG CB 4.03 C17 262-ARG CB 3.56 O3 262-ARG CB 3.55 O4 262-ARG CB 3.91 C17 262-ARG CD 4.09 O4 262-ARG CD 3.71 N1 263-ALA N 3.61 N1 263-ALA CA 3.59 N1 263-ALA CB3.54 O3 266-ARG NH1 3.93 N1 275-THR O 3.43 N1 276-LEU CA 3.46 N1 276-LEU C 3.83 C5 276-LEU CD1 4.02 C7 276-LEU CD2 4.00 C9 276-LEU CD2 3.81 C11 276-LEU CD2 3.91 C13 277-SER N 3.79 C15 277-SER N 3.63 C17 277-SER N 3.70 N1 277-SER N 3.17 O3277-SER N 3.37 C17 277-SER CA 3.89 O3 277-SER CA 3.43 C13 277-SER OG 3.66 O2 287-LEU CD1 4.05 C18 290-GLY C 4.04 C18 290-GLY O 3.48 C18 291-GLY CA 4.02 C4 292-LEU CG 3.89 C6 292-LEU CG 4.02 C2 292-LEU CD1 3.79 C4 292-LEU CD1 3.96 O2 292-LEUCD1 3.97 C4 292-LEU CD2 4.07 C6 292-LEU CD2 3.75 C8 292-LEU CD2 3.67 C10 292-LEU CD2 3.92 BR2 299-ILE CD1 3.68 C8 381-HIS CD2 3.92 C10 381-HIS CD2 3.78 O1 381-HIS GD2 3.50 O1 381-HIS CE1 3.62 C8 381-HIS NE2 3.36 C10 381-HIS NE2 3.34 O1381-HIS NE2 2.62 C8 401-PHE CE1 4.02 O1 401-PHE CE1 3.19 C16 401-PHE CZ 4.03 O1 401-PHE CZ 3.06 O3 502-H.sub.2 O O1 3.40 N1 502-H2O O1 3.12 O4 503-H.sub.2 O O1 3.20 C17 503-H2O O1 3.04 O3 503-H.sub.2 O O1 2.27 C15 504-H2O O1 4.01 C17504-H.sub.2 O O1 2.99 O3 504-H2O O1 3.80 O4 504-H.sub.2 O O1 1.78 Legend to Table 10. The table lists the interactions with IpBr2. The column headings are as follows: #1 The atom of IpBr2 that interacts with the amino acid of the receptor. Theseare also numbered in FIG. 32. #2 The amino acid in the full length rTR.alpha. that interacts with the ligand. #3 The name of the atom in the amino acid (standard nomenclature) where the interaction occurs. #4 The distance in A between IpBr2 and theprotein atom.

TABLE 11 Amino Acid Amino Acid Distance T3 Atom in full length .alpha. Atom A I2 215-PHE CD1 4.08 I1 218-PHE 0 3.19 I1 218-PHE CB 3.99 C4 218-PHE CB 4.04 I1 218-PHE C 3.79 I1 218-PHE CB 3.99 I1 221-ILE CG1 4.01 I1 222-ILE CB 3.95 I1222-ILE CG1 3.91 C8 222-ILE CD1 3.99 C10 222-ILE CD1 3.57 C12 222-ILE CD1 3.68 C13 225-ALA CB 3.66 C3 225-ALA CB 4.04 04 228-ARG NH1 3.23 04 228-ARG CZ 3.45 C15 228-ARG NH2 3.54 03 228-ARG NH2 3.90 04 228-ARG NH2 2.86 C10 256-MET SD 3.73 C12256-MET SD 3.90 C10 256-MET CE 3.97 C12 256-MET CE 3.92 I3 256-MET CE 3.89 C11 259-MET C 3.95 C11 259-MET 0 3.59 C14 259-MET 0 3.51 N1 259-MET 0 3.88 C1 259-MET CB 4.06 C11 259-MET CB 3.77 C13 259-MET CB 3.96 C15 262-ARG CB 3.61 C14 262-ARGCB 4.02 03 262-ARG CB 3.65 04 262-ARG CB 3.92 04 262-ARG CD 3.72 N1 263-ALA N 3.81 N1 263-ALA CA 3.81 N1 263-ALA CB 3.63 N1 275-THR 0 3.54 N1 276-LEU CA 3.38 N1 276-LEU C 3.73 C5 276-LEU CD1 4.00 C7 276-LEU CD1 4.05 02 276-LEU CD1 4.03 C7276-LEU CD2 3.80 C9 276-LEU CD2 3.70 C11 276-LEU CD2 4.01 C14 277-SER N 3.67 C15 277-SER N 3.62 N1 277-SER N 3.07 03 277-SER N 3.24 C15 277-SER CA 3.77 03 277-SER CA 3.34 C13 277-SER OG 3.92 12 290-GLY 0 3.50 C4 292-LEU CG 3.95 C8 292-LEU CG3.83 C2 292-LEU CD1 4.07 C4 292-LEU CD1 3.99 C4 292-LEU CD2 4.09 C6 292-LEU CD2 3.58 C8 292-LEU CD2 3.50 C10 292-LEU CD2 3.96 01 292-LEU CD2 3.71 I3 299-ILE CD1 3.74 C8 381-HIS CD2 3.94 C10 381-HIS CD2 3.97 01 381-HIS CD2 3.39 01 381-HIS CD13.82 C8 381-HIS NE2 3.47 C10 381-HIS NE2 3.55 01 381-HIS NE2 2.70 01 388-MET CE 3.88 01 401-PHE CE1 3.52 01 401-PHE CZ 3.32 C14 502-H20 01 4.01 C15 502-H20 01 3.61 03 502-H20 01 2.51 C15 503-H20 01 3.31 04 503-H.sub.2 0 01 3.10 N1 502-H.sub.20 01 3.27 03 503-H20 01 2.81 C15 504-H20 01 3.92 04 504-H20 01 2.73 Legend to Table 11. The table lists the interactions with T3. The column headings are as follows: #1 The atom of T3 that interacts with the amino acid of the receptor. These arealso numbered in FIG 32. #2 The amino acid in the full length rTR.alpha. that interacts with the ligand. #3 The name of the atom in the amino acid (standard nomenclature) where the interaction occurs. #4 The distance in A between T3 and the proteinatom.

TABLE 12 Triac Amino Acid Amino Acid Atom in full length hTR .beta. Atom Distance A I2 269-PHE CD1 3.75 I2 269-PHE CE1 3.88 I1 272-PHE C 4.03 I1 272-PHE 0 3.54 I1 275-ILE CG1 3.93 I1 276-ILE CG1 4.02 C3 279-ALA CB 3.81 C13 279-ALA CB3.87 C10 310-MET SD 3.72 C12 310-MET SD 3.78 C10 310-MET CE 4.02 C12 310-MET CE 3.92 I3 310-MET CE 3.93 C13 313-MET CA 3.94 C11 313-MET C 3.72 C1 313-MET 0 3.79 C11 313-MET 0 3.12 C13 313-MET 0 3.55 C1 313-MET CB 4.00 C11 313-MET CB 3.82 C13313-MET CB 3.76 C13 313-MET CG 3.88 03 316-ARG CB 3.99 04 317-ALA CA 4.08 04 317-ALA CA 4.10 C11 317-ALA CB 3.70 I3 317-ALA CB 4.10 04 317-ALA CB 4.06 04 320-ARG NH1 3.58 03 320-ARG NH2 3.55 04 320-ARG NH2 4.04 04 329-THR 0 3.55 04 330-LEU CA3.42 04 330-LEU C 3.77 C3 330-LEU CB 4.06 C5 330-LEU CB 4.08 C1 330-LEU CD2 4.07 C3 330-LEU CD2 4.00 C5 330-LEU CD2 3.73 C7 330-LEU CD2 3.51 C9 330-LEU CD2 3.54 C11 330-LEU CD2 3.86 C15 331-ASN N 3.55 03 331-ASN N 3.74 04 331-ASN N 3.12 03331-ASN CA 4.02 I2 344-GLY 0 3.87 C6 346-LEU CD2 3.87 C8 346-LEU CD2 3.84 01 346-LEU CD2 3.91 13 353-ILE CD1 3.51 C8 435-HIS CD2 3.93 C10 435-HIS CD2 3.79 01 435-HIS CD2 3.33 01 435-HIS CE1 3.81 C8 435-HIS NE2 3.42 C10 435-HIS NE2 3.33 01435-HIS NE2 2.67 01 442-MET SD 3.96 01 442-MET CE 3.72 I2 442-MET SD 4.01 01 455-PHE CE1 3.92 01 455-PHE CZ 3.50 Legend to Table 12. The table lists the interactions with Triac. The column headings are as follows: #1 The atom of Triac thatinteracts with the amino acid of the receptor. These are also numbered in FIG 32. #2 The amino acid in the full length hTR.beta. that interacts with the ligand. #3 The name of the atom in the amino acid (standard nomenclature) where the interactionoccurs. #4 The distance in A between Triac and the protein atom.

TABLE 13 GC1 Amino Acid Amino Acid Atom in full length TR .beta. Atom Distance A C16 269-PHE CE1 3.99 C19 272-PHE 0 3.85 C16 272-PHE CB 3.98 C16 273-THR CG2 3.76 C19 275-ILE CG1 3.98 C19 276-ILE CA 3.98 C2 276-ILE CD1 3.88 C8 276-ILECD1 3.77 C10 276-ILE CD1 3.58 C12 276-ILE CD1 3.62 C19 276-ILE CD1 3.56 C1 279-ALA CB 3.68 C3 279-ALA CB 3.56 05 279-ALA CB 3.11 04 279-ALA CB 3.90 03 282-ARG CZ 3.53 C17 282-ARG NH1 3.87 03 282-ARG NH1 3.20 04 282-ARG NH1 3.85 C17 282-ARGNH2 3.63 03 282-ARG NH2 3.00 C10 310-MET SD 3.86 C12 310-MET SD 3.91 C11 313-MET C 3.85 C11 313-MET 0 3.41 C15 313-MET 0 3.87 C20 313-MET 0 3.99 C11 313-MET CB 3.79 C1 313-MET CG 3.94 C11 313-MET CG 3.91 05 313-MET CG 3.87 04 313-MET CG 3.79 C20 314-SER CA 4.00 C17 316-ARG CB 3.95 C17 316-ARG CD 3.80 03 316-ARG CD 3.83 04 316-ARG CD 3.51 C20 317-ALA CB 3.93 C7 330-LEU CD2 3.56 C9 330-LEU CD2 3.63 C21 330-LEU CD2 3.90 05 331-ASN N 3.62 C15 331-ASN N 3.67 C18 344-GLY 0 3.60 C18346-LEU CG 3.89 C6 346-LEU CD2 3.77 C8 346-LEU CD2 3.80 C10 435-HIS CD2 3.89 01 435-HIS CD2 3.64 01 435-HIS CE1 3.79 C8 435-HIS NE2 3.44 C10 435-HIS NE2 3.33 01 435-HIS NE2 2.77 01 455-PHE CE1 3.40 01 455-PHE CZ 3.22 Legend to Table 13. Thetable lists the interactions with GC1. The column headings are as follows: #1 The atom of GC1 that interacts with the amino acid of the receptor. These are also numbered in FIG 32. #2 The amino acid in the full length hTR.beta. that interacts withthe ligand. #3 The name of the atom in the amino acid (standard nomenclature) where the interaction occurs. #4 The distance in A between GC1 and the protein atom.

TABLE 14 Coordination Structure of TR-.alpha. and Dimit R.sub.1 Coordination --CH.sub.2 -- R.sub.2 R.sub.3 R.sub.5 R.sub.6 R'.sub.2 R'.sub.3 R'.sub.4 R'.sub.5 R'.sub.6 X Structure CH(NH.sub.2)(CO.sub.2)H --H --CH.sub.3 --CH.sub.3 --H --H--CH(CH.sub.3).sub.2 --OH --H --H O AA 215 SS H3 AA 218 218 SS H3 H3 AA 219 SS H3 AA 221 SS H3 AA 222 222 222 222 SS H3 H3 H3 H3 AA 225 SS H3 AA 228 SS H3 AA 256 256 SS H5- H5- H6 H6 AA 259 259 SS H5-H6 H5-H6 AA 262 SS H5-H6 AA263 SS H5-H6 AA 266 SS loop AA 275 SS S3 AA 276 276 276 276 SS S3 S3 S3 S3 AA 277 SS loop AA 290-291 SS loop AA 292 292 292 292 292 SS loop loop loop loop loop AA 299 SS H8 AA 381 381 SS H11 H11 AA 388 SS H11 AA 401 401 SS H12 H12 AA H0H502/H0H5 03/H0H504 SS AA = Amino Acid SS = Secondary Structure

TABLE 15 Coordination Structure of TR-.alpha. and Triac Coordination R.sub.1 R.sub.2 R.sub.3 R.sub.5 R.sub.6 R'.sub.2 R'.sub.3 R'.sub.4 R'.sub.5 R'.sub.6 X Structure --CH.sub.2 --COOH --H --I --I --H --H --I --OH --H --H O AA 218 SS H3 AA 221 SS H3 AA 222 222 222 222 SS H3 H3 H3 H3 AA 225 SS H3 AA 256 256 256 SS H5-H6 H5-H6 H5-H6 AA 259 259 SS H5-H6 H5- H6 AA 262 SS H5-H6 AA 263 SS H5-H6 AA 266 SS loop AA 275 SS S3 AA 276 276 276 276 SS S3 S3 S3 S3 AA 277 SSloop AA 290 SS loop AA 292 292 292 292 292 SS loop loop loop loop loop AA 299 SS H8 AA 381 381 SS H11 H11 AA 388 SS H11 AA 401 401 SS H12 H12 AA = Amino Acid SS = Secondary Structure

TABLE 16 Coordination Structure of TR-.alpha. and IpBr2 Coordination R.sub.1 R.sub.2 R.sub.3 R.sub.5 R.sub.6 R'.sub.2 R'.sub.3 R'.sub.4 R'.sub.5 R'.sub.6 X Structure --CH.sub.2 --CH(NH.sub.2)(CO.sub.2)H --H --Br --Br --H --H--CH(CH.sub.3).sub.2 --OH --H --H O AA 215 SS H3 AA 218 218 SS H3 H3 AA 219 SS H3 AA 221 SS H3 AA 222 222 222 222 SS H3 H3 H3 H3 AA 225 SS H3 AA 228 SS H3 AA 256 256 256 SS H5-H6 H5-H6 H5- H6 AA 259 259 SS H5-H6 H5-H6 AA 262 SSH5-H6 AA 263 SS H5-H6 AA 266 SS loop AA 275 SS S3 AA 276 276 276 276 SS S3 S3 S3 S3 AA 277 SS AA 290- 291 SS loop AA 292 292 292 292 292 SS loop loop loop loop loop AA 299 SS H8 AA 381 381 SS H11 H11 AA 401 401 SS H12 H12 AAH0H502/H0H 503/H0H504 SS AA = Amino Acid SS = Secondary Structure

TABLE 17 Coordination Structure of TR-.alpha. and Dimit R.sub.1 Coordination --CH.sub.2 -- R.sub.2 R.sub.3 R.sub.5 R.sub.6 R'.sub.2 R'.sub.3 R'.sub.4 R'.sub.5 R'.sub.6 X Structure CH(NH.sub.2)(CO.sub.2)H --H --I --I --H --H --I --OH --H--H O AA 215 SS H3 AA 218 218 SS H3 H3 AA 221 SS H3 AA 222 222 222 222 SS H3 H3 H3 H3 AA 225 SS H3 AA 228 SS H3 AA 256 256 256 SS H5- H5- H5-H6 H6 H6 AA 259 259 SS H5-H6 H5- H6 AA 262 SS H5-H6 AA 263 SS H5-H6 AA 275 SS S3 AA276 276 276 276 SS S3 S3 S3 S3 AA 277 SS AA 290 SS loop AA 292 292 292 292 292 SS loop loop loop loop loop AA 299 SS H8 AA 381 381 SS H11 H11 AA 388 SS H11 AA 401 401 SS H12 H12 AA H0H502/H0H 503/H0H504 SS AA = Amino Acid SS =Secondary Structure

TABLE 18 Coordination Structure of TR-.beta. and Triac Coordination R1 R2 R3 R5 R6 R2' R3' R4' R5' R6' X Structure --CH.sub.2 CO.sub.2 H H I I H H I OH H H O AA 269 SS H3 AA 272 SS H3 AA 275 SS H3 AA 276 SS H3 AA 279 279 SS H3 H3 AA 310 310 310 SS H5-H6 H5-H6 H5-H6 AA 313 313 SS H5-H6 H5-H6 AA 316 SS H5-H6 AA 317 317 317 SS H5-H6 H5-H6 H5-H6 AA 320 SS H5-H6 AA 329 SS S3 AA 330 330 330 330 330 SS S3 S3 S3 S3 S3 AA 331 SS loop AA 344 SS loop AA 346 346 SS looploop AA 353 SS H8 AA 435 435 SS H11 H11 AA 442 442 SS H11 H11 AA 455 SS H12 AA = Amino Acid SS = Secondary Structure

TABLE 19 Coordination Structure of TR-.beta. and GC1 Coordination R.sub.1 R.sub.2 R.sub.3 R.sub.5 R.sub.6 R2' R3' R4' R5' R6' X Structure --O--CH.sub.2 CO.sub.2 H H CH.sub.3 CH.sub.3 H H CH(CH.sub.3) OH H H CH.sub.2 AA 269 SS H3 AA 272 SS H3 AA 273 273 SS H3 H3 AA 275 SS H3 AA 276 276 276 276 SS H3 H3 H3 H3 AA 279 279 SS H3 H3 AA 282 SS H3 AA 310 310 310 SS H5-H6 H5-H6 H5-H6 AA 313 313 SS H5-H6 H5-H6 AA 314 SS H5-H6 AA 316 SS H5-H6 AA 317 SS H5-H6 AA 320 SSH5-H6 AA 329 SS S3 AA 330 330 SS S3 S3 AA 331 SS loop AA 344 SS loop AA 346 346 SS loop loop AA 353 SS H8 AA 435 435 H11 H11 455 SS H12 AA = Amino Acid SS = Secondary Structure

APPENDIX 3 TR_DMT.PDB REMARK TR_full length numbering REMARK REMARK Rfactor 0.205 Rfree 0.227 REMARK Resolution 15. 2.2 all reflections REMARK REMARK Three cacodylate-modified cysteines (CYA) REMARK Cya334, Cya380, Cya392 REMARKcacodylate modeled as single arsenic atom REMARK REMARK side chain of certain residues modeled as ALA due to poor density; REMARK however, residue name reflects true residue for clarity REMARK REMARK clone obtained from Murray et. al. REMARKdeposited sequence confirmed, REMARK differing from that reported by Thompson et. al. REMARK in the following codons: REMARK 281 Thr--Ala REMARK 285 Lys--Glu REMARK identical to that reported by Mitsuhashi et. al. REMARK gb:RNTRAVI X07409 JRNLAUTH M. B. MURRAY, N. D. ZILZ, N. L. MCCREARY, M. J. MACDONALD JRNL AUTH 2 H. C. TOWLE JRNL TITL ISOLATION AND CHARACTERIZATION OF RAT CDNA CLONES FOR TWO JRNL TITL 2 DISTINCT THYROID HORMONE RECPTORS JRNL REF JBC V. 263 25 1988 JRNL AUTH C. C.THOMPSON, C. WEINBERGER, R. LEBO, R. M. EVANS JRNL TITL IDENTIFICATION OF A NOVEL THYROID HORMONE RECEPTOR EXPRESSED JRNL TITL 2 IN THE MAMMALIAN CENTRAL NERVOUS SYSTEM JRNL REF SCIENCE V. 237 1987 JRNL AUTH T. MITSUHASHI, G. TENNYSON, V. NIKODEM JRNL TITL NUCLEOTIDE SEQUENCE OF NOVEL CDNAS GENERATED BY ALTERNATIVE JRNL TITL 2 SPLICING OF A RAT THYROID HORMONE RECEPTOR GENE TRANSCRIPT JRNL REF NUC. ACIDS. RES. V. 16 12 1988 ATOM 1 N ARG 157 68.504 8.445 5.651 1.00 68.93 ATOM 2 CA ARG 15767.886 9.543 6.398 1.00 56.98 ATOM 3 CB ARG 157 68.769 10.789 6.324 1.00 59.25 ATOM 4 CG ARG 157 70.147 10.632 6.932 1.00 58.90 ATOM 5 CD ARG 157 70.068 10.422 8.425 1.00 59.37 ATOM 6 NE ARG 157 71.392 10.446 9.036 1.00 63.94 ATOM 7 CZ ARG 15771.613 10.329 10.341 1.00 64.39 ATOM 8 NH1 ARG 157 70.596 10.182 11.179 1.00 62.14 ATOM 9 NH2 ARG 157 72.855 10.365 10.808 1.00 65.56 ATOM 10 C ARG 157 66.500 9.881 5.854 1.00 48.97 ATOM 11 O ARG 157 66.351 10.203 4.674 1.00 48.61 ATOM 12 N PRO158 65.469 9.818 6.712 1.00 41.90 ATOM 13 CD PRO 158 65.550 9.366 8.112 1.00 41.06 ATOM 14 CA PRO 158 64.083 10.114 6.333 1.00 39.34 ATOM 15 CB PRO 158 63.286 9.704 7.576 1.00 37.89 ATOM 16 CG PRO 158 64.260 9.883 8.693 1.00 42.40 ATOM 17 C PRO158 63.814 11.573 5.930 1.00 37.10 ATOM 18 O PRO 158 64.189 12.517 6.636 1.00 33.31 ATOM 19 N GLU 159 63.171 11.733 4.778 1.00 30.56 ATOM 20 CA GLU 159 62.821 13.038 4.231 1.00 24.26 ATOM 21 CB GLU 159 62.553 12.904 2.727 1.00 19.19 ATOM 22 CGGLU 159 63.788 12.677 1.874 1.00 20.60 ATOM 23 CD GLU 159 64.407 13.971 1.390 1.00 26.54 ATOM 24 OE1 GLU 159 63.649 14.929 1.115 1.00 30.85 ATOM 25 OE2 GLU 159 65.649 14.027 1.268 1.00 28.35 ATOM 26 C GLU 159 61.549 13.520 4.909 1.00 23.26 ATOM27 O GLU 159 60.906 12.765 5.643 1.00 26.86 ATOM 28 N PRO 160 61.200 14.806 4.729 1.00 22.72 ATOM 29 CD PRO 160 61.981 15.916 4.153 1.00 17.87 ATOM 30 CA PRO 160 59.969 15.292 5.359 1.00 19.90 ATOM 31 CB PRO 160 60.004 16.799 5.070 1.00 14.42 ATOM 32 CG PRO 160 61.465 17.109 4.919 1.00 12.87 ATOM 33 C PRO 160 58.747 14.623 4.701 1.00 23.68 ATOM 34 O PRO 160 58.730 14.383 3.491 1.00 24.72 ATOM 35 N THR 161 57.749 14.281 5.506 1.00 22.19 ATOM 36 CA THR 161 56.542 13.660 4.985 1.00 19.50 ATOM 37 CB THR 161 55.691 13.031 6.125 1.00 21.50 ATOM 38 OG1 THR 161 55.163 14.062 6.972 1.00 20.33 ATOM 39 CG2 THR 161 56.537 12.078 6.959 1.00 19.48 ATOM 40 C THR 161 55.744 14.765 4.298 1.00 22.86 ATOM 41 O THR 161 56.040 15.949 4.4811.00 27.68 ATOM 42 N PRO 162 54.720 14.403 3.504 1.00 20.36 ATOM 43 CD PRO 162 54.280 13.050 3.113 1.00 16.55 ATOM 44 CA PRO 162 53.924 15.435 2.830 1.00 21.97 ATOM 45 CB PRO 162 52.780 14.633 2.210 1.00 18.17 ATOM 46 CG PRO 162 53.422 13.3161.905 1.00 18.01 ATOM 47 C PRO 162 53.399 16.467 3.826 1.00 22.56 ATOM 48 O PRO 162 53.461 17.675 3.567 1.00 21.73 ATOM 49 N GLU 163 52.912 15.976 4.967 1.00 25.28 ATOM 50 CA GLU 163 52.357 16.816 6.030 1.00 26.64 ATOM 51 CB GLU 163 51.74315.962 7.144 1.00 30.22 ATOM 52 CG GLU 163 50.514 15.131 6.748 1.00 44.99 ATOM 53 CD GLU 163 50.836 13.950 5.831 1.00 48.88 ATOM 54 OE1 GLU 163 50.016 13.660 4.929 1.00 52.48 ATOM 55 OE2 GLU 163 51.895 13.309 6.015 1.00 44.23 ATOM 56 C GLU 16353.414 17.731 6.634 1.00 27.65 ATOM 57 O GLU 163 53.114 18.862 7.034 1.00 29.30 ATOM 58 N GLU 164 54.646 17.235 6.712 1.00 21.89 ATOM 59 CA GLU 164 55.741 18.015 7.265 1.00 18.29 ATOM 60 CB GLU 164 56.901 17.109 7.657 1.00 14.78 ATOM 61 CG GLU164 56.552 16.196 8.825 1.00 21.11 ATOM 62 CD GLU 164 57.669 15.249 9.198 1.00 20.35 ATOM 63 OE1 GLU 164 58.605 15.071 8.392 1.00 28.55 ATOM 64 OE2 GLU 164 57.610 14.677 10.302 1.00 28.25 ATOM 65 C GLU 164 56.200 19.097 6.306 1.00 24.62 ATOM 66O GLU 164 56.574 20.183 6.741 1.00 32.05 ATOM 67 N TRP 165 56.174 18.817 5.003 1.00 28.22 ATOM 68 CA TRP 165 56.576 19.825 4.021 1.00 22.99 ATOM 69 CB TRP 165 56.575 19.262 2.605 1.00 17.37 ATOM 70 CG TRP 165 57.876 18.633 2.210 1.00 10.74 ATOM71 CD2 TRP 165 59.153 19.283 2.109 1.00 11.74 ATOM 72 CE2 TRP 165 60.075 18.319 1.648 1.00 9.97 ATOM 73 CE3 TRP 165 59.606 20.583 2.365 1.00 13.88 ATOM 74 CD1 TRP 165 58.074 17.343 1.832 1.00 9.17 ATOM 75 NE1 TRP 165 59.390 17.145 1.486 1.00 16.55 ATOM 76 CZ2 TRP 165 61.427 18.613 1.436 1.00 13.37 ATOM 77 CZ3 TRP 165 60.954 20.874 2.156 1.00 16.15 ATOM 78 CH2 TRP 165 61.846 19.892 1.696 1.00 17.42 ATOM 79 C TRP 165 55.634 21.015 4.115 1.00 21.44 ATOM 80 O TRP 165 56.041 22.149 3.8651.00 22.12 ATOM 81 N ASP 166 54.373 20.747 4.456 1.00 21.29 ATOM 82 CA ASP 166 53.369 21.796 4.621 1.00 25.77 ATOM 83 CB ASP 166 51.972 21.196 4.808 1.00 26.02 ATOM 84 CG ASP 166 51.428 20.559 3.539 1.00 33.01 ATOM 85 OD1 ASP 166 51.874 20.9322.434 1.00 29.48 ATOM 86 OD2 ASP 166 50.537 19.692 3.649 1.00 34.47 ATOM 87 C ASP 166 53.732 22.637 5.842 1.00 27.91 ATOM 88 O ASP 166 53.744 23.865 5.767 1.00 31.28 ATOM 89 N LEU 167 54.046 21.966 6.951 1.00 25.57 ATOM 90 CA LEU 167 54.43922.640 8.187 1.00 28.28 ATOM 91 CB LEU 167 54.854 21.624 9.256 1.00 32.80 ATOM 92 CG LEU 167 53.945 21.347 10.455 1.00 41.75 ATOM 93 CD1 LEU 167 54.765 20.640 11.532 1.00 39.15 ATOM 94 CD2 LEU 167 53.374 22.647 11.008 1.00 39.20 ATOM 95 C LEU167 55.636 23.532 7.902 1.00 22.19 ATOM 96 O LEU 167 55.671 24.700 8.302 1.00 29.51 ATOM 97 N ILE 168 56.610 22.957 7.206 1.00 15.01 ATOM 98 CA ILE 168 57.846 23.632 6.833 1.00 18.03 ATOM 99 CB ILE 168 58.756 22.668 6.040 1.00 11.37 ATOM 100CG2 ILE 168 59.890 23.413 5.367 1.00 16.36 ATOM 101 CG1 ILE 168 59.289 21.580 6.975 1.00 21.63 ATOM 102 CD1 ILE 168 60.095 20.501 6.287 1.00 21.03 ATOM 103 C ILE 168 57.579 24.897 6.022 1.00 22.54 ATOM 104 O ILE 168 58.155 25.948 6.300 1.00 24.88 ATOM 105 N HIS 169 56.682 24.800 5.045 1.00 25.70 ATOM 106 CA HIS 169 56.337 25.934 4.190 1.00 21.28

ATOM 107 CB HIS 169 55.411 25.493 3.057 1.00 22.29 ATOM 108 CG HIS 169 56.047 24.543 2.091 1.00 23.11 ATOM 109 CD2 HIS 169 57.348 24.265 1.839 1.00 16.86 ATOM 110 ND1 HIS 169 55.312 23.721 1.263 1.00 25.30 ATOM 111 CE1 HIS 169 56.13022.974 0.546 1.00 15.89 ATOM 112 NE2 HIS 169 57.371 23.283 0.878 1.00 25.38 ATOM 113 C HIS 169 55.664 27.048 4.976 1.00 18.32 ATOM 114 O HIS 169 56.033 28.215 4.842 1.00 21.53 ATOM 115 N VAL 170 54.679 26.685 5.795 1.00 17.13 ATOM 116 CA VAL170 53.957 27.661 6.607 1.00 21.29 ATOM 117 CB VAL 170 52.808 26.991 7.399 1.00 24.33 ATOM 118 CG1 VAL 170 52.164 27.985 8.354 1.00 23.78 ATOM 119 CG2 VAL 170 51.760 26.439 6.435 1.00 18.87 ATOM 120 C VAL 170 54.910 28.382 7.567 1.00 24.69 ATOM121 O VAL 170 54.912 29.616 7.637 1.00 28.77 ATOM 122 N ALA 171 55.759 27.609 8.245 1.00 20.35 ATOM 123 CA ALA 171 56.722 28.148 9.202 1.00 19.61 ATOM 124 CB ALA 171 57.393 27.013 9.977 1.00 17.52 ATOM 125 C ALA 171 57.775 29.026 8.531 1.00 20.91 ATOM 126 O ALA 171 58.102 30.105 9.041 1.00 21.98 ATOM 127 N THR 172 58.308 28.571 7.398 1.00 18.94 ATOM 128 CA THR 172 59.313 29.342 6.668 1.00 19.55 ATOM 129 CB THR 172 59.820 28.594 5.413 1.00 20.49 ATOM 130 OG1 THR 172 60.394 27.3365.795 1.00 20.66 ATOM 131 CG2 THR 172 60.894 29.418 4.702 1.00 20.44 ATOM 132 C THR 172 58.730 30.697 6.254 1.00 23.26 ATOM 133 O THR 172 59.403 31.724 6.334 1.00 24.32 ATOM 134 N GLU 173 57.468 30.694 5.836 1.00 27.42 ATOM 135 CA GLU 17356.797 31.922 5.434 1.00 27.68 ATOM 136 CB GLU 173 55.477 31.605 4.728 1.00 24.51 ATOM 137 CG GLU 173 54.652 32.836 4.338 1.00 39.69 ATOM 138 CD GLU 173 55.396 33.814 3.426 1.00 47.72 ATOM 139 OE1 GLU 173 55.019 35.009 3.417 1.00 48.26 ATOM 140OE2 GLU 173 56.344 33.398 2.717 1.00 49.61 ATOM 141 C GLU 173 56.557 32.834 6.641 1.00 25.68 ATOM 142 O GLU 173 56.773 34.046 6.559 1.00 23.39 ATOM 143 N ALA 174 56.119 32.245 7.755 1.00 25.19 ATOM 144 CA ALA 174 55.863 32.989 8.993 1.00 22.25 ATOM 145 CB ALA 174 55.450 32.030 10.111 1.00 15.95 ATOM 146 C ALA 174 57.125 33.747 9.391 1.00 23.22 ATOM 147 O ALA 174 57.076 34.918 9.768 1.00 24.52 ATOM 148 N HIS 175 58.261 33.073 9.275 1.00 20.97 ATOM 149 CA HIS 175 59.544 33.665 9.606 1.00 19.55 ATOM 150 CB HIS 175 60.625 32.577 9.649 1.00 16.19 ATOM 151 CG HIS 175 62.016 33.104 9.835 1.00 18.89 ATOM 152 CD2 HIS 175 63.148 32.901 9.119 1.00 16.05 ATOM 153 ND1 HIS 175 62.359 33.962 10.859 1.00 13.83 ATOM 154 CE1 HIS 175 63.64234.265 10.765 1.00 15.87 ATOM 155 NE2 HIS 175 64.143 33.635 9.718 1.00 19.19 ATOM 156 C HIS 175 59.934 34.757 8.617 1.00 21.28 ATOM 157 O HIS 175 60.274 35.869 9.014 1.00 25.12 ATOM 158 N ARG 176 59.891 34.436 7.329 1.00 26.73 ATOM 159 CA ARG176 60.266 35.387 6.292 1.00 27.13 ATOM 160 CB ARG 176 60.156 34.748 4.914 1.00 36.00 ATOM 161 CG ARG 176 61.286 33.795 4.602 1.00 43.20 ATOM 162 CD ARG 176 61.197 33.334 3.170 1.00 50.07 ATOM 163 NE ARG 176 62.316 32.477 2.813 1.00 58.20 ATOM164 CZ ARG 176 62.266 31.548 1.867 1.00 67.22 ATOM 165 NH ARG 176 61.143 31.358 1.182 1.00 67.62 ATOM 166 NH2 ARG 176 63.336 30.806 1.612 1.00 70.56 ATOM 167 C ARG 176 59.487 36.688 6.325 1.00 23.97 ATOM 168 O ARG 176 60.073 37.760 6.209 1.00 24.52 ATOM 169 N SER 177 58.177 36.598 6.515 1.00 23.60 ATOM 170 CA SER 177 57.341 37.789 6.565 1.00 26.36 ATOM 171 CB SER 177 55.865 37.407 6.439 1.00 21.93 ATOM 172 OG SER 177 55.495 36.459 7.423 1.00 25.97 ATOM 173 C SER 177 57.557 38.6237.829 1.00 28.76 ATOM 174 O SER 177 57.084 39.761 7.907 1.00 33.09 ATOM 175 N THR 178 58.257 38.062 8.815 1.00 25.52 ATOM 176 CA THR 178 58.508 38.772 10.064 1.00 18.93 ATOM 177 CB THR 178 57.828 38.064 11.258 1.00 21.81 ATOM 178 OG1 THR 17858.348 36.736 11.394 1.00 24.18 ATOM 179 CG2 THR 178 56.330 37.971 11.032 1.00 13.81 ATOM 180 C THR 178 59.993 38.967 10.358 1.00 20.69 ATOM 181 O THR 178 60.373 39.407 11.448 1.00 20.56 ATOM 182 N ASN 179 60.837 38.645 9.385 1.00 23.68 ATOM183 CA ASN 179 62.275 38.802 9.555 1.00 28.22 ATOM 184 CB ASN 179 63.022 37.627 8.927 1.00 27.45 ATOM 185 CG ASN 179 64.460 37.529 9.402 1.00 33.98 ATOM 186 OD1 ASN 179 65.342 37.131 8.644 1.00 42.72 ATOM 187 ND2 ASN 179 64.702 37.865 10.667 1.00 31.14 ATOM 188 C ASN 179 62.689 40.115 8.902 1.00 34.47 ATOM 189 O ASN 179 62.832 40.200 7.678 1.00 36.54 ATOM 190 N ALA 180 62.874 41.135 9.735 1.00 37.39 ATOM 191 CA ALA 180 63.235 42.479 9.292 1.00 33.71 ATOM 192 CB ALA 180 63.555 43.35210.494 1.00 31.57 ATOM 193 C ALA 180 64.375 42.545 8.284 1.00 37.87 ATOM 194 O ALA 180 65.458 42.018 8.525 1.00 35.26 ATOM 195 N GLN 181 64.095 43.187 7.150 1.00 40.55 ATOM 196 CA GLN 181 65.049 43.391 6.057 1.00 42.95 ATOM 197 CB GLN 18166.344 44.043 6.570 1.00 45.47 ATOM 198 CG GLN 181 66.144 45.326 7.383 1.00 52.70 ATOM 199 CD GLN 181 65.351 46.399 6.650 1.00 55.03 ATOM 200 OE1 GLN 181 65.270 46.412 5.421 1.00 59.56 ATOM 201 NE2 GLN 181 64.757 47.308 7.411 1.00 54.39 ATOM202 C GLN 181 65.391 42.176 5.197 1.00 44.27 ATOM 203 O GLN 181 66.181 42.291 4.251 1.00 46.47 ATOM 204 N GLY 182 64.797 41.025 5.508 1.00 42.17 ATOM 205 CA GLY 182 65.054 39.815 4.742 1.00 42.63 ATOM 206 C GLY 182 66.522 39.584 4.427 1.00 47.40 ATOM 207 O GLY 182 67.382 39.691 5.306 1.00 49.38 ATOM 208 N SER 183 66.816 39.297 3.163 1.00 49.46 ATOM 209 CA SER 183 68.189 39.061 2.733 1.00 54.13 ATOM 210 CB SER 183 68.208 38.225 1.449 1.00 55.08 ATOM 211 OG SER 183 67.197 38.647 0.546 1.00 63.54 ATOM 212 C SER 183 68.949 40.369 2.532 1.00 54.84 ATOM 213 O SER 183 70.175 40.373 2.407 1.00 56.90 ATOM 214 N HIS 184 68.223 41.482 2.535 1.00 55.77 ATOM 215 CA HIS 184 68.854 42.775 2.342 1.00 57.78 ATOM 216 C HIS 184 69.605 43.296 3.5561.00 59.09 ATOM 217 O HIS 184 70.312 44.301 3.454 1.00 60.34 ATOM 218 N TRP 185 69.502 42.597 4.686 1.00 55.60 ATOM 219 CA TRP 185 70.159 43.020 5.923 1.00 53.73 ATOM 220 CB TRP 185 69.973 41.973 7.030 1.00 50.40 ATOM 221 CG TRP 185 70.74640.694 6.837 1.00 48.09 ATOM 222 CD2 TRP 185 72.091 40.419 7.269 1.00 47.38 ATOM 223 CE2 TRP 185 72.390 39.094 6.888 1.00 40.29 ATOM 224 CE3 TRP 185 73.071 41.169 7.937 1.00 45.43 ATOM 225 CD1 TRP 185 70.301 39.554 6.234 1.00 49.87 ATOM 226 NE1TRP 185 71.280 38.589 6.262 1.00 48.02 ATOM 227 CZ2 TRP 185 73.628 38.496 7.154 1.00 38.65 ATOM 228 CZ3 TRP 185 74.304 40.573 8.201 1.00 43.26 ATOM 229 CH2 TRP 185 74.570 39.250 7.807 1.00 40.00 ATOM 230 C TRP 185 71.638 43.386 5.800 1.00 55.99 ATOM 231 O TRP 185 72.089 44.359 6.401 1.00 52.84 ATOM 232 N LYS 186 72.389 42.614 5.021 1.00

59.15 ATOM 233 CA LYS 186 73.818 42.863 4.843 1.00 64.01 ATOM 234 CB LYS 186 74.466 41.688 4.091 1.00 64.67 ATOM 235 CG LYS 186 75.943 41.868 3.729 1.00 65.58 ATOM 236 CD LYS 186 76.817 42.181 4.946 1.00 62.03 ATOM 237 CE LYS 186 78.23842.512 4.515 1.00 61.52 ATOM 238 NZ LYS 186 78.988 43.243 5.579 1.00 61.67 ATOM 239 C LYS 186 74.131 44.203 4.160 1.00 67.49 ATOM 240 O LYS 186 75.164 44.816 4.432 1.00 68.66 ATOM 241 N GLN 187 73.221 44.678 3.316 1.00 68.99 ATOM 242 CA GLN 18773.431 45.939 2.612 1.00 69.65 ATOM 243 CB GLN 187 72.880 45.867 1.180 1.00 73.76 ATOM 244 CG GLN 187 73.632 44.935 0.237 1.00 78.61 ATOM 245 CD GLN 187 73.368 43.471 0.525 1.00 84.96 ATOM 246 OE1 GLN 187 74.203 42.782 1.109 1.00 87.73 ATOM 247NE2 GLN 187 72.197 42.989 0.122 1.00 84.98 ATOM 248 C GLN 187 72.817 47.141 3.323 1.00 69.16 ATOM 249 O GLN 187 73.379 48.235 3.299 1.00 71.39 ATOM 250 N ARG 188 71.666 46.936 3.953 1.00 65.82 ATOM 251 CA ARG 188 70.961 48.014 4.639 1.00 65.00 ATOM 252 CB ARG 188 69.458 47.739 4.591 1.00 66.20 ATOM 253 CG ARG 188 68.957 47.483 3.181 1.00 70.30 ATOM 254 CD ARG 188 67.463 47.212 3.132 1.00 78.59 ATOM 255 NE ARG 188 67.003 47.008 1.760 1.00 87.71 ATOM 256 CZ ARG 188 67.011 47.946 0.8141.00 94.10 ATOM 257 NH1 ARG 188 67.453 49.171 1.081 1.00 97.26 ATOM 258 NH2 ARG 188 66.589 47.657 -0.409 1.00 94.07 ATOM 259 C ARG 188 71.409 48.286 6.077 1.00 65.39 ATOM 260 O ARG 188 70.900 49.201 6.727 1.00 65.20 ATOM 261 N ARG 189 72.37247.506 6.561 1.00 64.28 ATOM 262 CA ARG 189 72.882 47.654 7.922 1.00 60.75 ATOM 263 CB ARG 189 73.691 46.409 8.321 1.00 56.87 ATOM 264 CG ARG 189 75.050 46.308 7.630 1.00 59.52 ATOM 265 CD ARG 189 75.580 44.891 7.589 1.00 55.86 ATOM 266 NE ARG189 75.874 44.348 8.907 1.00 55.48 ATOM 267 CZ ARG 189 77.055 43.849 9.257 1.00 61.38 ATOM 268 NH1 ARG 189 78.057 43.832 8.388 1.00 62.54 ATOM 269 NH2 ARG 189 77.225 43.328 10.465 1.00 62.20 ATOM 270 C ARG 189 73.747 48.907 8.082 1.00 60.91 ATOM 271 O ARG 189 74.548 49.245 7.207 1.00 60.67 ATOM 272 N LYS 190 73.575 49.591 9.207 1.00 59.06 ATOM 273 CA LYS 190 74.340 50.790 9.521 1.00 55.00 ATOM 274 CB LYS 190 73.423 52.008 9.582 1.00 55.45 ATOM 275 C LYS 190 74.991 50.542 10.875 1.00 51.52 ATOM 276 O LYS 190 74.320 50.144 11.830 1.00 51.68 ATOM 277 N PHE 191 76.304 50.721 10.944 1.00 50.49 ATOM 278 CA PHE 191 77.037 50.508 12.186 1.00 50.17 ATOM 279 CB PHE 191 78.546 50.571 11.943 1.00 48.38 ATOM 280 CG PHE 191 79.090 49.42311.142 1.00 49.66 ATOM 281 CD1 PHE 191 78.873 49.348 9.768 1.00 51.03 ATOM 282 CD2 PHE 191 79.845 48.429 11.759 1.00 46.28 ATOM 283 CE1 PHE 191 79.403 48.298 9.018 1.00 51.35 ATOM 284 CE2 PHE 191 80.379 47.377 11.021 1.00 47.26 ATOM 285 CZ PHE191 80.158 47.311 9.646 1.00 48.48 ATOM 286 C PHE 191 76.663 51.534 13.248 1.00 48.61 ATOM 287 O PHE 191 76.507 52.720 12.952 1.00 50.38 ATOM 288 N LEU 192 76.488 51.068 14.479 1.00 47.31 ATOM 289 CA LEU 192 76.169 51.958 15.584 1.00 42.72 ATOM290 CB LEU 192 75.845 51.151 16.844 1.00 36.66 ATOM 291 CG LEU 192 75.397 51.949 18.068 1.00 31.01 ATOM 292 CD1 LEU 192 74.048 52.590 17.786 1.00 28.37 ATOM 293 CD2 LEU 192 75.318 51.043 19.289 1.00 29.60 ATOM 294 C LEU 192 77.447 52.760 15.8001.00 42.28 ATOM 295 O LEU 192 78.528 52.179 15.932 1.00 39.71 ATOM 296 N PRO 193 77.350 54.104 15.781 1.00 45.15 ATOM 297 CD PRO 193 76.095 54.865 15.617 1.00 43.82 ATOM 298 CA PRO 193 78.493 55.006 15.973 1.00 43.14 ATOM 299 CB PRO 193 77.82056.306 16.400 1.00 44.37 ATOM 300 CG PRO 193 76.571 56.308 15.565 1.00 41.66 ATOM 301 C PRO 193 79.476 54.498 17.028 1.00 43.34 ATOM 302 O PRO 193 79.103 54.296 18.182 1.00 45.18 ATOM 303 N ASP 194 80.732 54.317 16.628 1.00 44.22 ATOM 304 CAASP 194 81.781 53.804 17.512 1.00 47.20 ATOM 305 CB ASP 194 83.108 53.732 16.761 1.00 41.89 ATOM 306 C ASP 194 81.962 54.511 18.866 1.00 51.99 ATOM 307 O ASP 194 82.636 53.986 19.752 1.00 54.04 ATOM 308 N ASP 195 81.381 55.698 19.025 1.00 55.21 ATOM 309 CA ASP 195 81.489 56.428 20.288 1.00 57.50 ATOM 310 CB ASP 195 81.423 57.948 20.061 1.00 60.04 ATOM 311 CG ASP 195 80.123 58.398 19.406 1.00 68.39 ATOM 312 OD1 ASP 195 79.211 58.847 20.136 1.00 69.46 ATOM 313 OD2 ASP 195 80.020 58.32218.162 1.00 72.91 ATOM 314 C ASP 195 80.410 55.976 21.280 1.00 58.05 ATOM 315 O ASP 195 80.540 56.180 22.491 1.00 58.97 ATOM 316 N ILE 196 79.349 55.363 20.759 1.00 56.06 ATOM 317 CA ILE 196 78.247 54.863 21.580 1.00 50.48 ATOM 318 CB ILE 19676.930 54.762 20.766 1.00 45.82 ATOM 319 CG2 ILE 196 75.818 54.166 21.621 1.00 44.04 ATOM 320 CG1 ILE 196 76.517 56.147 20.261 1.00 44.27 ATOM 321 CD1 ILE 196 75.179 56.171 19.541 1.00 45.25 ATOM 322 C ILE 196 78.603 53.484 22.435 1.00 47.66 ATOM 323 O ILE 196 79.138 52.636 21.419 1.00 43.96 ATOM 324 N GLY 197 78.309 53.269 23.414 1.00 46.29 ATOM 325 CA GLY 197 78.608 51.995 24.045 1.00 48.03 ATOM 326 C GLY 197 79.978 51.963 24.692 1.00 50.42 ATOM 327 O GLY 197 80.463 50.902 25.0701.00 46.66 ATOM 328 N GLN 198 80.583 53.137 24.854 1.00 56.94 ATOM 329 CA GLN 198 81.910 53.259 25.454 1.00 59.51 ATOM 330 CB GLN 198 82.751 54.257 24.649 1.00 62.53 ATOM 331 CG GLN 198 83.232 53.718 23.316 1.00 69.39 ATOM 332 CD GLN 198 84.08852.484 23.483 1.00 76.76 ATOM 333 OE1 GLN 198 83.745 51.399 22.996 1.00 81.73 ATOM 334 NE2 GLN 198 85.205 52.632 24.192 1.00 78.09 ATOM 335 C GLN 198 81.915 53.678 26.922 1.00 57.56 ATOM 336 O GLN 198 82.946 53.584 27.588 1.00 57.71 ATOM 337 NSER 199 80.770 54.128 27.425 1.00 54.11 ATOM 338 CA SER 199 80.676 54.600 28.800 1.00 46.28 ATOM 339 CB SER 199 80.243 56.067 28.777 1.00 50.28 ATOM 340 OG SER 199 80.935 56.776 27.757 1.00 50.95 ATOM 341 C SER 199 79.776 53.805 29.757 1.00 40.19 ATOM 342 O SER 199 78.680 54.252 30.102 1.00 39.26 ATOM 343 N PRO 200 80.236 52.629 30.214 1.00 35.63 ATOM 344 CD PRO 200 81.530 52.011 29.904 1.00 34.88 ATOM 345 CA PRO 200 79.464 51.789 31.139 1.00 37.54 ATOM 346 CB PRO 200 80.223 50.45731.124 1.00 29.86 ATOM 347 CG PRO 200 81.207 50.570 29.995 1.00 34.29 ATOM 348 C PRO 200 79.521 52.416 32.532 1.00 44.63 ATOM 349 O PRO 200 80.443 52.137 33.300 1.00 47.80 ATOM 350 N ILE 201 78.532 53.241 32.867 1.00 49.57 ATOM 351 CA ILE 20178.525 53.924 34.158 1.00 49.15 ATOM 352 CB ILE 201 78.213 55.426 33.990 1.00 49.19 ATOM 353 CG2 ILE 201 78.429 56.150 35.306 1.00 53.37 ATOM 354 CG1 ILE 201 79.137 56.037 32.934 1.00 52.55 ATOM 355 CD1 ILE 201 78.811 57.471 32.586 1.00 55.26 ATOM 356 C ILE 201 77.625 53.352 35.254 1.00 49.88 ATOM 357 O ILE 201 78.044 53.250 36.408 1.00 50.20

ATOM 358 N VAL 202 76.384 53.014 34.920 1.00 47.85 ATOM 359 CA VAL 202 75.468 52.474 35.927 1.00 45.76 ATOM 360 CB VAL 202 74.015 52.415 35.400 1.00 39.98 ATOM 361 CG1 VAL 202 73.072 51.896 36.482 1.00 35.94 ATOM 362 CG2 VAL 202 73.57453.799 34.944 1.00 29.43 ATOM 363 C VAL 202 75.954 51.093 36.373 1.00 50.57 ATOM 364 O VAL 202 76.296 50.249 35.545 1.00 49.50 ATOM 365 N SER 203 76.009 50.876 37.683 1.00 54.82 ATOM 366 CA SER 203 76.490 49.609 38.223 1.00 59.26 ATOM 367 CBSER 203 77.067 49.809 39.628 1.00 64.88 ATOM 368 OG SER 203 76.127 50.428 40.492 1.00 75.47 ATOM 369 C SER 203 75.457 48.491 38.244 1.00 55.78 ATOM 370 O SER 203 74.285 48.712 38.544 1.00 57.50 ATOM 371 N MET 204 75.923 47.283 37.958 1.00 52.29 ATOM 372 CA MET 204 75.076 46.103 37.948 1.00 50.42 ATOM 373 CB MET 204 75.032 45.487 36.548 1.00 47.74 ATOM 374 CG MET 204 74.243 46.297 35.541 1.00 43.40 ATOM 375 SD MET 204 72.491 46.348 35.953 1.00 40.93 ATOM 376 CE MET 204 71.947 44.78535.241 1.00 39.19 ATOM 377 C MET 204 75.670 45.107 38.925 1.00 49.42 ATOM 378 O MET 204 76.892 45.020 39.062 1.00 52.25 ATOM 379 N PRO 205 74.816 44.329 39.605 1.00 47.73 ATOM 380 CD PRO 205 73.344 44.414 39.549 1.00 48.94 ATOM 381 CA PRO 20575.250 43.326 40.580 1.00 47.34 ATOM 382 CB PRO 205 73.982 42.513 40.810 1.00 49.44 ATOM 383 CG PRO 205 72.907 43.562 40.725 1.00 50.62 ATOM 384 C PRO 205 76.431 42.442 40.168 1.00 47.12 ATOM 385 O PRO 205 77.299 42.160 40.990 1.00 51.21 ATOM386 N ASP 206 76.487 42.023 38.909 1.00 48.81 ATOM 387 CA ASP 206 77.583 41.160 38.465 1.00 49.88 ATOM 388 CB ASP 206 77.128 40.223 37.330 1.00 54.06 ATOM 389 CG ASP 206 76.598 40.967 36.107 1.00 57.34 ATOM 390 OD1 ASP 206 77.056 42.095 35.8111.00 52.21 ATOM 391 OD2 ASP 206 75.719 40.397 35.423 1.00 59.16 ATOM 392 C ASP 206 78.902 41.843 38.093 1.00 48.70 ATOM 393 O ASP 206 79.862 41.171 37.715 1.00 49.75 ATOM 394 N GLY 207 78.946 43.168 38.161 1.00 47.54 ATOM 395 CA GLY 207 80.17443.869 37.820 1.00 49.23 ATOM 396 C GLY 207 80.169 44.585 36.482 1.00 51.96 ATOM 397 O GLY 207 80.783 45.645 36.348 1.00 56.32 ATOM 398 N ASP 208 79.510 44.005 35.481 1.00 52.50 ATOM 399 CA ASP 208 79.435 44.624 34.157 1.00 48.00 ATOM 400 CBASP 208 78.968 43.609 33.115 1.00 53.23 ATOM 401 CG ASP 208 80.038 42.592 32.774 1.00 53.17 ATOM 402 OD1 ASP 208 81.130 43.006 32.335 1.00 57.42 ATOM 403 OD2 ASP 208 79.787 41.380 32.942 1.00 55.64 ATOM 404 C ASP 208 78.497 45.823 34.187 1.00 46.68 ATOM 405 O ASP 208 77.283 45.671 34.332 1.00 45.81 ATOM 406 N LYS 209 79.075 47.014 34.077 1.00 45.95 ATOM 407 CA LYS 209 78.313 48.257 34.115 1.00 45.87 ATOM 408 CB LYS 209 79.235 49.418 34.478 1.00 46.90 ATOM 409 C LYS 209 77.561 48.54632.812 1.00 41.17 ATOM 410 O LYS 209 77.951 48.074 31.745 1.00 39.51 ATOM 411 N VAL 210 76.500 49.344 32.916 1.00 39.35 ATOM 412 CA VAL 210 75.652 49.713 31.782 1.00 38.03 ATOM 413 CB VAL 210 74.136 49.584 32.140 1.00 32.13 ATOM 414 CG1 VAL 21073.269 49.926 30.937 1.00 27.92 ATOM 415 CG2 VAL 210 73.818 48.183 32.627 1.00 29.43 ATOM 416 C VAL 210 75.895 51.134 31.263 1.00 38.68 ATOM 417 O VAL 210 76.090 52.079 32.038 1.00 39.57 ATOM 418 N ASP 211 75.848 51.272 29.942 1.00 39.19 ATOM419 CA ASP 211 76.019 52.544 29.254 1.00 38.39 ATOM 420 CB ASP 211 76.794 52.327 27.946 1.00 40.36 ATOM 421 CG ASP 211 77.051 53.620 27.177 1.00 36.85 ATOM 422 OD4 ASP 211 76.193 54.528 27.167 1.00 37.95 ATOM 423 OD2 ASP 211 78.121 53.716 26.5531.00 33.87 ATOM 424 C ASP 211 74.601 53.040 28.958 1.00 40.60 ATOM 425 O ASP 211 73.919 52.517 28.073 1.00 40.36 ATOM 426 N LEU 212 74.185 54.074 29.680 1.00 41.55 ATOM 427 CA LEU 212 72.854 54.664 29.552 1.00 38.39 ATOM 428 CB LEU 212 72.75955.883 30.467 1.00 40.93 ATOM 429 CG LEU 212 71.575 55.979 31.428 1.00 45.32 ATOM 430 CD1 LEU 212 71.271 54.626 32.047 1.00 43.83 ATOM 431 CD2 LEU 212 71.900 57.007 32.502 1.00 44.93 ATOM 432 C LEU 212 72.448 55.050 28.133 1.00 37.61 ATOM 433 OLEU 212 71.318 54.805 27.719 1.00 33.71 ATOM 434 N GLU 213 73.360 55.670 27.393 1.00 41.23 ATOM 435 CA GLU 213 73.068 56.984 26.023 1.00 43.48 ATOM 436 CB GLU 213 74.181 56.986 25.481 1.00 47.66 ATOM 437 CG GLU 213 73.919 57.494 24.065 1.00 56.87 ATOM 438 CD GLU 213 75.121 58.180 23.433 1.00 60.87 ATOM 439 OE1 GLU 213 76.258 57.996 23.924 1.00 60.37 ATOM 440 OE2 GLU 213 74.921 58.894 22.423 1.00 61.13 ATOM 441 C GLU 213 72.889 54.880 25.102 1.00 39.29 ATOM 442 O GLU 213 71.96554.841 24.290 1.00 36.66 ATOM 443 N ALA 214 73.785 53.906 25.233 1.00 36.33 ATOM 444 CA ALA 214 73.739 52.693 24.422 1.00 34.89 ATOM 445 CB ALA 214 74.946 51.817 24.711 1.00 30.70 ATOM 446 C ALA 214 72.454 51.938 24.718 1.00 31.96 ATOM 447 OALA 214 71.739 51.523 23.804 1.00 33.93 ATOM 448 N PHE 215 72.151 51.798 26.003 1.00 28.47 ATOM 449 CA PHE 215 70.947 51.116 26.445 1.00 29.74 ATOM 450 CB PHE 215 70.819 51.223 27.962 1.00 23.73 ATOM 451 CG PHE 215 69.589 50.568 28.515 1.00 22.71 ATOM 452 CD1 PHE 215 69.603 49.220 28.858 1.00 22.53 ATOM 453 CD2 PHE 215 68.423 51.301 28.712 1.00 19.74 ATOM 454 CE1 PHE 215 68.477 48.606 29.391 1.00 20.75 ATOM 455 CE2 PHE 215 67.290 50.698 29.245 1.00 21.02 ATOM 456 CZ PHE 215 67.31849.346 29.586 1.00 19.50 ATOM 457 C PHE 215 69.730 51.742 25.771 1.00 34.64 ATOM 458 O PHE 215 68.872 51.034 25.239 1.00 39.86 ATOM 459 N SER 216 69.677 53.071 25.771 1.00 34.78 ATOM 460 CA SER 216 68.572 53.801 25.160 1.00 36.01 ATOM 461 CBSER 216 68.762 55.302 25.366 1.00 37.36 ATOM 462 OG SER 216 67.537 55.987 25.193 1.00 48.33 ATOM 463 C SER 216 68.458 53.475 23.664 1.00 37.06 ATOM 464 O SER 216 67.358 53.250 23.148 1.00 33.23 ATOM 465 N GLU 217 69.601 53.410 22.986 1.00 36.25 ATOM 466 CA GLU 217 69.645 53.091 21.562 1.00 36.99 ATOM 467 CB GLU 217 71.092 53.104 21.064 1.00 37.10 ATOM 468 CG GLU 217 71.682 54.491 20.912 1.00 44.30 ATOM 469 CD GLU 217 71.016 55.284 19.802 1.00 51.30 ATOM 470 OE1 GLU 217 71.439 55.14218.633 1.00 57.25 ATOM 471 OE2 GLU 217 70.070 56.046 20.096 1.00 52.50 ATOM 472 C GLU 217 69.019 51.726 21.286 1.00 36.93 ATOM 473 O GLU 217 68.191 51.577 20.381 1.00 41.06 ATOM 474 N PHE 218 69.395 50.740 22.093 1.00 30.27 ATOM 475 CA PHE 21868.875 49.388 21.947 1.00 27.20 ATOM 476 CB PHE 218 69.679 48.421 22.814 1.00 28.10 ATOM 477 CG PHE 218 71.124 48.330 22.428 1.00 24.84 ATOM 478 CD1 PHE 218 72.117 48.286 23.398 1.00 21.78 ATOM 479 CD2 PHE 218 71.495 48.301 21.087 1.00 24.78 ATOM 480 CE1 PHE 218 73.458 48.215 23.040 1.00 24.08 ATOM 481 CE2 PHE 218 72.834 48.230 20.719 1.00 25.33 ATOM 482 CZ PHE 218 73.818 48.187 21.697 1.00 25.04 ATOM 483 C PHE 218 67.381 49.281 22.261 1.00

28.23 ATOM 484 O PHE 218 66.639 48.605 21.543 1.00 33.52 ATOM 485 N THR 219 66.927 49.961 23.310 1.00 27.24 ATOM 486 CA THR 219 65.515 49.913 23.666 1.00 29.28 ATOM 487 CB THR 219 65.238 50.533 25.052 1.00 30.97 ATOM 488 OG1 THR 21965.724 51.880 25.090 1.00 35.50 ATOM 489 CG2 THR 219 65.901 49.712 26.149 1.00 30.78 ATOM 490 C THR 219 64.660 50.612 22.615 1.00 33.29 ATOM 491 O THR 219 63.473 50.317 22.474 1.00 36.85 ATOM 492 N LYS 220 65.276 51.515 21.860 1.00 35.23 ATOM493 CA LYS 220 64.579 52.253 20.816 1.00 38.97 ATOM 494 CB LYS 220 65.506 53.334 20.236 1.00 44.67 ATOM 495 CG LYS 220 64.805 54.491 19.513 1.00 58.02 ATOM 496 CD LYS 220 64.406 54.130 18.079 1.00 68.57 ATOM 497 CE LYS 220 63.732 55.296 17.3471.00 70.50 ATOM 498 NZ LYS 220 62.395 55.668 17.905 1.00 66.08 ATOM 499 C LYS 220 64.112 51.289 19.721 1.00 38.48 ATOM 500 O LYS 220 63.021 51.446 19.173 1.00 37.18 ATOM 501 N ILE 221 64.917 50.270 19.432 1.00 36.19 ATOM 502 CA ILE 221 64.56349.305 18.394 1.00 36.77 ATOM 503 CB ILE 221 65.756 48.996 17.457 1.00 34.41 ATOM 504 CG2 ILE 221 66.270 50.276 16.814 1.00 38.54 ATOM 505 CG1 ILE 221 66.864 48.267 18.221 1.00 32.93 ATOM 506 CD1 ILE 221 67.984 47.752 17.338 1.00 31.12 ATOM 507C ILE 221 64.002 47.971 18.888 1.00 38.22 ATOM 508 O ILE 221 63.499 47.181 18.089 1.00 38.90 ATOM 509 N ILE 222 64.048 47.719 20.191 1.00 35.75 ATOM 510 CA ILE 222 63.557 46.446 20.702 1.00 31.77 ATOM 511 CB ILE 222 64.086 46.152 22.130 1.00 33.14 ATOM 512 CG2 ILE 222 63.203 46.813 23.183 1.00 24.60 ATOM 513 CG1 ILE 222 64.147 44.638 22.350 1.00 32.60 ATOM 514 CD1 ILE 222 64.860 44.226 23.609 1.00 34.52 ATOM 515 C ILE 222 62.042 46.240 20.624 1.00 32.56 ATOM 516 O ILE 222 61.58145.109 20.452 1.00 35.74 ATOM 517 N THR 223 61.262 47.313 20.720 1.00 29.43 ATOM 518 CA THR 223 59.806 47.170 20.651 1.00 33.57 ATOM 519 CB THR 223 59.075 48.514 20.903 1.00 38.99 ATOM 520 OG1 THR 223 59.422 49.010 22.205 1.00 41.23 ATOM 521CG2 THR 223 57.558 48.325 20.836 1.00 36.98 ATOM 522 C THR 223 59.355 46.528 19.325 1.00 31.45 ATOM 523 O THR 223 58.571 45.571 19.334 1.00 26.77 ATOM 524 N PRO 224 59.824 47.054 18.173 1.00 31.35 ATOM 525 CD PRO 224 60.570 48.306 17.950 1.00 30.11 ATOM 526 CA PRO 224 59.424 46.462 16.891 1.00 30.38 ATOM 527 CB PRO 224 60.149 47.336 15.865 1.00 30.09 ATOM 528 CG PRO 224 60.200 48.659 16.530 1.00 31.86 ATOM 529 C PRO 224 59.882 45.007 16.795 1.00 29.51 ATOM 530 O PRO 224 59.147 44.15316.295 1.00 32.52 ATOM 531 N ALA 225 61.090 44.734 17.285 1.00 22.63 ATOM 532 CA ALA 225 61.650 43.385 17.268 1.00 20.88 ATOM 533 CB ALA 225 63.046 43.386 17.862 1.00 20.57 ATOM 534 C ALA 225 60.752 42.416 18.026 1.00 23.53 ATOM 535 O ALA 22560.455 41.323 17.544 1.00 25.07 ATOM 536 N ILE 226 60.296 42.828 19.202 1.00 22.61 ATOM 537 CA ILE 226 59.420 41.989 20.007 1.00 19.46 ATOM 538 CB ILE 226 59.120 42.644 21.360 1.00 20.25 ATOM 539 CG2 ILE 226 58.071 41.843 22.105 1.00 16.75 ATOM540 CG1 ILE 226 60.401 42.772 22.182 1.00 19.30 ATOM 541 CD1 ILE 226 60.240 43.645 23.413 1.00 20.92 ATOM 542 C ILE 226 58.112 41.768 19.251 1.00 21.28 ATOM 543 O ILE 226 57.553 40.670 19.256 1.00 23.75 ATOM 544 N THR 227 57.629 42.821 18.5981.00 24.46 ATOM 545 CA THR 227 56.393 42.752 17.826 1.00 25.81 ATOM 546 CB THR 227 56.020 44.136 17.260 1.00 31.00 ATOM 547 OG1 THR 227 55.772 45.039 18.345 1.00 35.43 ATOM 548 CG2 THR 227 54.776 44.049 16.388 1.00 29.01 ATOM 549 C THR 22756.508 41.728 16.691 1.00 22.85 ATOM 550 O THR 227 55.589 40.939 16.469 1.00 22.84 ATOM 551 N ARG 228 57.647 41.713 16.004 1.00 16.09 ATOM 552 CA ARG 228 57.862 40.765 14.919 1.00 16.97 ATOM 553 CB ARG 228 59.161 41.064 14.174 1.00 14.71 ATOM554 CG ARG 228 59.137 42.369 13.391 1.00 16.22 ATOM 555 CD ARG 228 60.309 42.447 12.422 1.00 20.90 ATOM 556 NE ARG 228 61.595 42.207 13.078 1.00 24.94 ATOM 557 CZ ARG 228 62.243 43.113 13.805 1.00 35.06 ATOM 558 NH1 ARG 228 61.729 44.328 13.9731.00 36.35 ATOM 559 NH2 ARG 228 63.404 42.807 14.370 1.00 32.78 ATOM 560 C ARG 228 57.866 39.326 15.431 1.00 21.63 ATOM 561 O ARG 228 57.477 38.407 14.704 1.00 24.47 ATOM 562 N VAL 229 58.304 39.128 16.675 1.00 20.00 ATOM 563 CA VAL 229 58.31937.793 17.266 1.00 18.39 ATOM 564 CB VAL 229 59.103 37.745 18.606 1.00 19.20 ATOM 565 CG1 VAL 229 58.938 36.382 19.265 1.00 14.19 ATOM 566 CG2 VAL 229 60.581 38.001 18.356 1.00 14.81 ATOM 567 C VAL 229 56.875 37.367 17.501 1.00 20.00 ATOM 568 OVAL 229 56.499 36.227 17.212 1.00 20.04 ATOM 569 N VAL 230 56.058 38.291 18.003 1.00 19.60 ATOM 570 CA VAL 230 54.651 37.996 18.247 1.00 18.72 ATOM 571 CB VAL 230 53.930 39.185 18.912 1.00 22.15 ATOM 572 CG1 VAL 230 52.452 38.862 19.113 1.00 15.66 ATOM 573 CG2 VAL 230 54.592 39.522 20.248 1.00 21.05 ATOM 574 C VAL 230 53.967 37.660 16.917 1.00 26.17 ATOM 575 O VAL 230 53.188 36.704 16.836 1.00 28.01 ATOM 576 N ASP 231 54.288 38.426 15.873 1.00 25.07 ATOM 577 CA ASP 231 53.714 38.21614.542 1.00 26.10 ATOM 578 CB ASP 231 54.169 39.309 13.568 1.00 22.15 ATOM 579 CG ASP 231 53.620 40.684 13.921 1.00 29.49 ATOM 580 OD1 ASP 231 52.587 40.767 14.624 1.00 30.93 ATOM 581 OD2 ASP 231 54.223 41.687 13.481 1.00 31.74 ATOM 582 C ASP231 54.087 36.842 13.989 1.00 27.35 ATOM 583 O ASP 231 53.245 36.154 13.408 1.00 25.89 ATOM 584 N PHE 232 55.347 36.451 14.175 1.00 24.29 ATOM 585 CA PHE 232 55.825 35.154 13.714 1.00 22.90 ATOM 586 CB PHE 232 57.302 34.956 14.090 1.00 20.56 ATOM 587 CG PHE 232 57.762 33.525 14.007 1.00 24.20 ATOM 588 CD1 PHE 232 57.952 32.910 12.772 1.00 23.44 ATOM 589 CD2 PHE 232 57.959 32.776 15.167 1.00 19.41 ATOM 590 CE1 PHE 232 58.329 31.567 12.689 1.00 19.53 ATOM 591 CE2 PHE 232 58.336 31.43115.100 1.00 21.09 ATOM 592 CZ PHE 232 58.520 30.824 13.858 1.00 21.61 ATOM 593 C PHE 232 54.984 34.047 14.341 1.00 24.18 ATOM 594 O PHE 232 54.481 33.160 13.645 1.00 22.26 ATOM 595 N ALA 233 54.810 34.127 15.656 1.00 23.90 ATOM 596 CA ALA 23354.048 33.128 16.397 1.00 22.60 ATOM 597 CB ALA 233 54.088 33.435 17.890 1.00 15.34 ATOM 598 C ALA 233 52.609 33.040 15.917 1.00 22.04 ATOM 599 O ALA 233 52.084 31.948 15.697 1.00 22.86 ATOM 600 N LYS 234 51.978 34.195 15.743 1.00 25.04 ATOM601 CA LYS 234 50.593 34.248 15.298 1.00 27.68 ATOM 602 CB LYS 234 50.096 35.691 15.292 1.00 31.41 ATOM 603 CG LYS 234 49.845 36.248 16.682 1.00 40.37 ATOM 604 CD LYS 234 49.212 37.626 16.604 1.00 57.53 ATOM 605 CE LYS 234 48.772 38.112 17.9741.00 64.28 ATOM 606 NZ LYS 234 48.164 39.473 17.904 1.00 67.19 ATOM 607 C LYS 234 50.358 33.588 13.939 1.00 26.42 ATOM 608 O LYS 234 49.269 33.067 13.674 1.00 31.34

ATOM 609 N LYS 235 51.382 33.588 13.093 1.00 24.38 ATOM 610 CA LYS 235 51.278 32.985 11.770 1.00 26.42 ATOM 611 CB LYS 235 52.244 33.664 10.805 1.00 24.92 ATOM 612 CG LYS 235 51.908 35.127 10.583 1.00 22.41 ATOM 613 CD LYS 235 52.84335.775 9.588 1.00 29.38 ATOM 614 CE LYS 235 52.481 37.234 9.395 1.00 33.49 ATOM 615 NZ LYS 235 53.354 37.869 8.376 1.00 40.13 ATOM 616 C LYS 235 51.470 31.469 11.759 1.00 30.02 ATOM 617 O LYS 235 51.417 30.838 10.699 1.00 30.37 ATOM 618 N LEU236 51.722 30.889 12.930 1.00 32.39 ATOM 619 CA LEU 236 51.878 29.443 13.053 1.00 36.24 ATOM 620 CB LEU 236 52.944 29.080 14.089 1.00 29.91 ATOM 621 CG LEU 236 54.373 29.516 13.765 1.00 24.69 ATOM 622 CD1 LEU 236 55.299 29.054 14.877 1.00 22.71 ATOM 623 CD2 LEU 236 54.811 28.942 12.427 1.00 24.48 ATOM 624 C LEU 236 50.520 28.891 13.470 1.00 41.22 ATOM 625 O LEU 236 49.936 29.333 14.467 1.00 41.45 ATOM 626 N PRO 237 50.012 27.895 12.729 1.00 47.86 ATOM 627 CD PRO 237 50.739 27.190 11.6571.00 49.32 ATOM 628 CA PRO 237 48.713 27.262 12.992 1.00 50.28 ATOM 629 CB PRO 237 48.669 26.128 11.962 1.00 55.25 ATOM 630 CG PRO 237 50.135 25.818 11.706 1.00 54.08 ATOM 631 C PRO 237 48.495 26.751 14.422 1.00 47.94 ATOM 632 O PRO 237 47.53327.134 15.087 1.00 42.48 ATOM 633 N MET 238 49.415 25.927 14.906 1.00 49.51 ATOM 634 CA MET 238 49.306 25.354 16.245 1.00 53.49 ATOM 635 CB MET 238 50.379 24.275 16.424 1.00 52.52 ATOM 636 CG MET 238 50.028 22.959 15.728 1.00 56.00 ATOM 637 SDMET 238 51.443 21.961 15.204 1.00 50.16 ATOM 638 CE MET 238 50.896 21.440 13.552 1.00 55.71 ATOM 639 C MET 238 49.352 26.362 17.395 1.00 54.20 ATOM 640 O MET 238 48.930 26.058 18.515 1.00 54.72 ATOM 641 N PHE 239 49.803 27.578 17.101 1.00 50.11 ATOM 642 CA PHE 239 49.917 28.619 18.117 1.00 41.11 ATOM 643 CB PHE 239 51.089 29.552 17.788 1.00 34.80 ATOM 644 CG PHE 239 51.336 30.607 18.826 1.00 30.25 ATOM 645 CD1 PHE 239 52.127 30.332 19.937 1.00 25.66 ATOM 646 CD2 PHE 239 50.786 31.87818.690 1.00 26.30 ATOM 647 CE1 PHE 239 52.368 31.307 20.896 1.00 30.28 ATOM 648 CE2 PHE 239 51.019 32.862 19.644 1.00 30.49 ATOM 649 CZ PHE 239 51.813 32.576 20.750 1.00 29.00 ATOM 650 C PHE 239 48.647 29.434 18.337 1.00 35.65 ATOM 651 O PHE239 48.151 29.521 19.457 1.00 30.27 ATOM 652 N SER 240 48.133 30.037 17.272 1.00 36.49 ATOM 653 CA SER 240 46.936 30.866 17.359 1.00 36.37 ATOM 654 CB SER 240 46.622 31.466 15.994 1.00 35.87 ATOM 655 C SER 240 45.707 30.145 17.936 1.00 40.37 ATOM 656 O SER 240 44.784 30.789 18.438 1.00 37.47 ATOM 657 N GLU 241 45.713 28.814 17.889 1.00 43.00 ATOM 658 CA GLU 241 44.605 28.004 18.404 1.00 46.31 ATOM 659 CB GLU 241 44.714 26.566 17.881 1.00 55.84 ATOM 660 CG GLU 241 44.750 26.422 16.3601.00 69.03 ATOM 661 CD GLU 241 45.141 25.015 15.900 1.00 74.99 ATOM 662 OE1 GLU 241 45.835 24.299 16.658 1.00 77.81 ATOM 663 OE2 GLU 241 44.765 24.629 14.770 1.00 70.58 ATOM 664 C GLU 241 44.587 27.961 19.933 1.00 42.60 ATOM 665 O GLU 24143.541 27.740 20.545 1.00 43.23 ATOM 666 N LEU 242 45.762 28.125 20.535 1.00 39.31 ATOM 667 CA LEU 242 45.926 28.086 21.987 1.00 34.54 ATOM 668 CB LEU 242 47.417 28.109 22.344 1.00 28.35 ATOM 669 CG LEU 242 48.311 26.974 21.853 1.00 27.59 ATOM670 CD1 LEU 242 49.750 27.307 22.180 1.00 20.72 ATOM 671 CD2 LEU 242 47.902 25.661 22.500 1.00 24.97 ATOM 672 C LEU 242 45.242 29.240 22.711 1.00 32.23 ATOM 673 O LEU 242 44.956 30.282 22.119 1.00 31.50 ATOM 674 N PRO 243 44.954 29.060 24.0101.00 34.39 ATOM 675 CD PRO 243 45.118 27.843 24.827 1.00 31.68 ATOM 676 CA PRO 243 44.309 30.134 24.773 1.00 34.39 ATOM 677 CB PRO 243 44.092 29.498 26.154 1.00 32.34 ATOM 678 CG PRO 243 44.081 28.026 25.892 1.00 33.80 ATOM 679 C PRO 243 45.30031.303 24.873 1.00 35.56 ATOM 680 O PRO 243 46.517 31.082 24.897 1.00 34.99 ATOM 681 N CYS 244 44.791 32.532 24.946 1.00 34.23 ATOM 682 CA CYS 244 45.648 33.714 25.062 1.00 37.03 ATOM 683 CB CYS 244 44.820 34.960 25.376 1.00 43.49 ATOM 684 SGCYS 244 43.820 35.531 24.007 1.00 71.28 ATOM 685 C CYS 244 46.716 33.555 26.135 1.00 34.99 ATOM 686 O CYS 244 47.894 33.802 25.882 1.00 37.49 ATOM 687 N GLU 245 46.305 33.125 27.326 1.00 33.03 ATOM 688 CA GLU 245 47.249 32.944 28.424 1.00 35.72 ATOM 689 CB GLU 245 46.559 32.469 29.71.6 1.00 37.85 ATOM 690 CG GLU 245 45.294 31.633 29.549 1.00 46.81 ATOM 691 CD GLU 245 44.029 32.478 29.480 1.00 44.81 ATOM 692 OE1 GLU 245 43.606 33.012 30.527 1.00 33.05 ATOM 693 OE2 GLU 245 43.454 32.59928.377 1.00 48.22 ATOM 694 C GLU 245 48.414 32.035 28.047 1.00 32.29 ATOM 695 O GLU 245 49.558 32.319 28.399 1.00 35.92 ATOM 696 N ASP 246 48.134 30.975 27.295 1.00 30.64 ATOM 697 CA ASP 246 49.182 30.058 26.855 1.00 28.23 ATOM 698 CB ASP 24648.575 28.809 26.208 1.00 30.51 ATOM 699 CG ASP 246 48.213 27.737 27.222 1.00 33.18 ATOM 700 OD1 ASP 246 48.265 28.006 28.439 1.00 31.26 ATOM 701 OD2 ASP 246 47.884 26.613 26.796 1.00 33.85 ATOM 702 C ASP 246 50.104 30.757 25.860 1.00 30.10 ATOM 703 O ASP 246 51.330 30.651 25.950 1.00 27.08 ATOM 704 N GLN 247 49.500 31.477 24.918 1.00 30.39 ATOM 705 CA GLN 247 50.249 32.208 23.901 1.00 29.08 ATOM 706 CB GLN 247 49.295 32.949 22.964 1.00 27.34 ATOM 707 CG GLN 247 48.390 32.034 22.1471.00 28.95 ATOM 708 CD GLN 247 47.531 32.796 21.153 1.00 30.74 ATOM 709 OE1 GLN 247 47.850 33.918 20.767 1.00 33.23 ATOM 710 NE2 GLN 247 46.439 32.185 20.729 1.00 35.19 ATOM 711 C GLN 247 51.190 33.196 24.575 1.00 27.51 ATOM 712 O GLN 24752.377 33.261 24.256 1.00 28.70 ATOM 713 N ILE 248 50.661 33.921 25.552 1.00 27.81 ATOM 714 CA ILE 248 51.431 34.908 26.295 1.00 29.41 ATOM 715 CB ILE 248 50.525 35.662 27.303 1.00 28.96 ATOM 716 CG2 ILE 248 51.356 36.476 28.279 1.00 28.67 ATOM717 CG1 ILE 248 49.555 36.571 26.543 1.00 28.83 ATOM 718 CD1 ILE 248 48.514 37.236 27.420 1.00 30.76 ATOM 719 C ILE 248 52.618 34.259 27.006 1.00 28.39 ATOM 720 O ILE 248 53.759 34.715 26.869 1.00 27.88 ATOM 721 N ILE 249 52.356 33.177 27.7321.00 26.07 ATOM 722 CA ILE 249 53.413 32.474 28.454 1.00 27.37 ATOM 723 CB ILE 249 52.839 31.294 29.281 1.00 30.32 ATOM 724 CG2 ILE 249 53.958 30.425 29.840 1.00 31.29 ATOM 725 CG1 ILE 249 51.987 31.831 30.429 1.00 30.31 ATOM 726 CD1 ILE 24951.295 30.753 31.230 1.00 31.30 ATOM 727 C ILE 249 54.510 31.974 27.509 1.00 28.63 ATOM 728 O ILE 249 55.701 32.100 27.808 1.00 29.59 ATOM 729 N LEU 250 54.110 31.442 26.357 1.00 29.03 ATOM 730 CA LEU 250 55.068 30.934 25.380 1.00 22.44 ATOM731 CB LEU 250 54.351 30.166 24.266 1.00 24.30 ATOM 732 CG LEU 250 53.665 28.866 24.687 1.00 23.20 ATOM 733 CD1 LEU 250 52.951 28.273 23.502 1.00 20.36 ATOM 734 CD2 LEU 250 54.685 27.880 25.238 1.00

19.45 ATOM 735 C LEU 250 55.919 32.055 24.794 1.00 18.97 ATOM 736 O LEU 250 57.133 31.903 24.648 1.00 18.37 ATOM 737 N LEU 251 55.291 33.180 24.468 1.00 20.63 ATOM 738 CA LEU 251 56.026 34.3.18 23.915 1.00 27.43 ATOM 739 CB LEU 25155.065 35.412 23.449 1.00 22.92 ATOM 740 CG LEU 251 54.364 35.093 22.128 1.00 24.72 ATOM 741 CD1 LEU 251 53.342 36.167 21.821 1.00 32.13 ATOM 742 CD2 LEU 251 55.389 34.981 21.009 1.00 22.46 ATOM 743 C LEU 251 57.026 34.875 24.930 1.00 27.23 ATOM 744 O LEU 251 58.202 35.078 24.614 1.00 26.48 ATOM 745 N LYS 252 56.561 35.094 26.156 1.00 27.34 ATOM 746 CA LYS 252 57.425 35.598 27.215 1.00 28.95 ATOM 747 CB LYS 252 56.649 35.715 28.527 1.00 32.89 ATOM 748 CG LYS 252 55.570 36.783 28.5301.00 35.06 ATOM 749 CD LYS 252 55.084 37.028 29.943 1.00 42.82 ATOM 750 CE LYS 252 54.124 38.191 30.003 1.00 53.05 ATOM 751 NZ LYS 252 53.677 38.451 31.398 1.00 64.03 ATOM 752 C LYS 252 58.605 34.647 27.405 1.00 27.66 ATOM 753 O LYS 252 59.73435.076 27.646 1.00 33.16 ATOM 754 N GLY 253 58.344 33.357 27.243 1.00 24.50 ATOM 755 CA GLY 253 59.386 32.364 27.402 1.00 22.33 ATOM 756 C GLY 253 60.423 32.273 26.297 1.00 23.99 ATOM 757 O GLY 253 61.589 32.016 26.581 1.00 30.77 ATOM 758 N CYS254 60.041 32.526 25.049 1.00 22.66 ATOM 759 CA CYS 254 60.986 32.405 23.934 1.00 20.75 ATOM 760 CB CYS 254 60.386 31.494 22.868 1.00 24.86 ATOM 761 SG CYS 254 58.996 32.276 22.014 1.00 25.55 ATOM 762 C CYS 254 61.399 33.702 23.242 1.00 23.79 ATOM 763 O CYS 254 62.262 33.685 22.357 1.00 22.18 ATOM 764 N CYS 255 60.788 34.814 23.625 1.00 19.49 ATOM 765 CA CYS 255 61.084 36.085 22.981 1.00 21.08 ATOM 766 CB CYS 255 60.336 37.220 23.669 1.00 18.21 ATOM 767 SG CYS 255 60.264 38.713 22.6771.00 22.96 ATOM 768 C CYS 255 62.570 36.413 22.842 1.00 21.87 ATOM 769 O CYS 255 63.050 36.641 21.729 1.00 22.23 ATOM 770 N MET 256 63.310 36.397 23.947 1.00 20.82 ATOM 771 CA MET 256 64.741 36.706 23.895 1.00 20.50 ATOM 772 CB MET 256 65.32236.801 25.312 1.00 22.50 ATOM 773 CG MET 256 66.808 37.139 25.354 1.00 16.67 ATOM 774 SD MET 256 67.205 38.732 24.605 1.00 24.46 ATOM 775 CE MET 256 69.027 38.764 24.791 1.00 19.21 ATOM 776 C MET 256 65.510 35.667 23.072 1.00 18.38 ATOM 777 OMET 256 66.401 36.005 22.293 1.00 17.68 ATOM 778 N GLU 257 65.149 34.404 23.248 1.00 20.33 ATOM 779 CA GLU 257 65.779 33.308 22.526 1.00 21.08 ATOM 780 CB GLU 257 65.148 31.982 22.943 1.00 22.28 ATOM 781 CG GLU 257 65.374 31.640 24.411 1.00 34.68 ATOM 782 CD GLU 257 64.515 30.486 24.907 1.00 43.20 ATOM 783 OE1 GLU 257 63.823 29.836 24.091 1.00 42.14 ATOM 784 OE2 GLU 257 64.530 30.230 26.128 1.00 50.15 ATOM 785 C GLU 257 65.650 33.503 21.018 1.00 19.26 ATOM 786 O GLU 257 66.63233.360 20.276 1.00 18.09 ATOM 787 N ILE 258 64.446 33.850 20.566 1.00 16.30 ATOM 788 CA ILE 258 64.199 34.065 19.141 1.00 18.09 ATOM 789 CB ILE 258 62.677 34.150 18.825 1.00 18.61 ATOM 790 CG2 ILE 258 62.441 34.653 17.395 1.00 16.23 ATOM 791CG1 ILE 258 62.032 32.771 19.021 1.00 13.80 ATOM 792 CD1 ILE 258 60.544 32.714 18.695 1.00 13.21 ATOM 793 C ILE 258 64.948 35.297 18.638 1.00 20.12 ATOM 794 O ILE 258 65.605 35.242 17.593 1.00 19.17 ATOM 795 N MET 259 64.903 36.387 19.404 1.00 22.71 ATOM 796 CA MET 259 65.602 37.611 19.015 1.00 17.09 ATOM 797 CB MET 259 65.249 38.772 19.941 1.00 18.80 ATOM 798 CG MET 259 63.782 39.159 19.894 1.00 17.66 ATOM 799 SD MET 259 63.457 40.748 20.678 1.00 25.77 ATOM 800 CE MET 259 63.77440.377 22.374 1.00 16.65 ATOM 801 C MET 259 67.111 37.397 18.973 1.00 19.51 ATOM 802 O MET 259 67.797 37.913 18.080 1.00 25.53 ATOM 803 N SER 260 67.625 36.605 19.908 1.00 19.58 ATOM 804 CA SER 260 69.056 36.324 19.947 1.00 16.90 ATOM 805 CBSER 260 69.434 35.631 21.251 1.00 15.56 ATOM 806 OG SER 260 69.093 36.455 22.352 1.00 22.98 ATOM 807 C SER 260 69.471 35.487 18.746 1.00 14.52 ATOM 808 O SER 260 70.496 35.761 18.129 1.00 22.82 ATOM 809 N LEU 261 68.663 34.490 18.397 1.00 16.50 ATOM 810 CA LEU 261 68.948 33.642 17.241 1.00 17.78 ATOM 811 CB LEU 261 67.878 32.552 17.092 1.00 18.38 ATOM 812 CG LEU 261 67.890 31.708 15.812 1.00 14.47 ATOM 813 CD1 LEU 261 69.159 30.877 15.728 1.00 16.76 ATOM 814 CD2 LEU 261 66.672 30.80615.793 1.00 14.06 ATOM 815 C LEU 261 68.959 34.519 15.992 1.00 20.40 ATOM 816 O LEU 261 69.885 34.450 15.181 1.00 22.00 ATOM 817 N ARG 262 67.934 35.356 15.854 1.00 21.02 ATOM 818 CA ARG 262 67.821 36.249 14.705 1.00 22.84 ATOM 819 CB ARG 26266.530 37.067 14.782 1.00 20.29 ATOM 820 CG ARG 262 65.311 36.267 14.364 1.00 23.33 ATOM 821 CD ARG 262 64.007 37.026 14.509 1.00 19.05 ATOM 822 NE ARG 262 62.959 36.321 13.775 1.00 21.32 ATOM 823 CZ ARG 262 61.780 36.837 13.441 1.00 23.44 ATOM824 NH1 ARG 262 61.465 38.081 13.780 1.00 22.99 ATOM 825 NH2 ARG 262 60.933 36.116 12.713 1.00 22.09 ATOM 826 C ARG 262 69.035 37.154 14.561 1.00 22.66 ATOM 827 O ARG 262 69.434 37.483 13.445 1.00 22.41 ATOM 828 N ALA 263 69.625 37.545 15.6891.00 23.52 ATOM 829 CA ALA 263 70.820 38.386 15.677 1.00 22.37 ATOM 830 CB ALA 263 70.986 39.089 17.018 1.00 22.76 ATOM 831 C ALA 263 72.052 37.530 15.366 1.00 22.85 ATOM 832 O ALA 263 72.882 37.897 14.529 1.00 25.50 ATOM 833 N ALA 264 72.13136.365 16.005 1.00 21.68 ATOM 834 CA ALA 264 73.242 35.438 15.826 1.00 20.26 ATOM 835 CB ALA 264 73.092 34.256 16.763 1.00 15.97 ATOM 836 C ALA 264 73.401 34.957 14.382 1.00 23.11 ATOM 837 O ALA 264 74.523 34.831 13.892 1.00 24.87 ATOM 838 NVAL 265 72.293 34.679 13.697 1.00 22.94 ATOM 839 CA VAL 265 72.380 34.226 12.306 1.00 28.98 ATOM 840 CB VAL 265 71.072 33.547 11.797 1.00 25.97 ATOM 841 CG1 VAL 265 70.751 32.330 12.638 1.00 26.27 ATOM 842 CG2 VAL 265 69.907 34.527 11.797 1.00 26.64 ATOM 843 C VAL 265 72.761 35.373 11.369 1.00 28.81 ATOM 844 O VAL 265 72.966 35.160 10.176 1.00 31.92 ATOM 845 N ARG 266 72.830 36.587 11.915 1.00 31.83 ATOM 846 CA ARG 266 73.210 37.774 11.150 1.00 33.19 ATOM 847 CB ARG 266 72.141 38.86111.258 1.00 31.67 ATOM 848 CG ARG 266 70.986 38.623 10.320 1.00 26.82 ATOM 849 CD ARG 266 69.913 39.668 10.454 1.00 33.95 ATOM 850 NE ARG 266 68.955 39.532 9.361 1.00 38.15 ATOM 851 CZ ARG 26.6 67.688 39.927 9.410 1.00 37.39 ATOM 852 NH1 ARG266 67.198 40.491 10.509 1.00 29.92 ATOM 853 NH2 ARG 266 66.918 39.770 8.340 1.00 31.24 ATOM 854 C ARG 266 74.565 38.307 11.604 1.00 36.31 ATOM 855 O ARG 266 74.821 39.516 11.575 1.00 38.56 ATOM 856 N TYR 267 75.416 37.393 12.056 1.00 34.21 ATOM 857 CA TYR 267 76.755 37.733 12.502 1.00 35.24 ATOM 858 CB TYR 267 77.283 36.640 13.440 1.00 32.37 ATOM 859 CG TYR 267 78.774 36.699 13.703 1.00 35.07

ATOM 860 CD1 TYR 267 79.303 37.555 14.669 1.00 33.94 ATOM 861 CE1 TYR 267 80.677 37.609 14.905 1.00 36.60 ATOM 862 CD2 TYR 267 79.658 35.894 12.979 1.00 34.68 ATOM 863 CE2 TYR 267 81.029 35.940 13.208 1.00 36.07 ATOM 864 CZ TYR 267 81.53336.797 14.170 1.00 37.14 ATOM 865 OH TYR 267 82.889 36.835 14.396 1.00 41.52 ATOM 866 C TYR 267 77.639 37.831 11.263 1.00 37.68 ATOM 867 O TYR 267 77.609 36.943 10.410 1.00 36.48 ATOM 868 N ASP 268 78.400 38.915 11.150 1.00 39.58 ATOM 869 CAASP 268 79.301 39.096 10.016 1.00 42.77 ATOM 870 CB ASP 268 79.170 40.511 9.434 1.00 44.38 ATOM 871 CG ASP 268 80.145 40.770 8.290 1.00 50.31 ATOM 872 OD1 ASP 268 80.290 39.901 7.400 1.00 55.79 ATOM 873 OD2 ASP 268 80.773 41.847 8.280 1.00 50.24 ATOM 874 C ASP 268 80.737 38.846 10.466 1.00 42.51 ATOM 875 O ASP 268 81.305 39.645 11.208 1.00 42.7S ATOM 876 N PRO 269 81.346 37.733 10.020 1.00 44.56 ATOM 877 CD PRO 269 80.770 36.697 9.146 1.00 42.66 ATOM 878 CA PRO 269 82.725 37.395 10.3931.00 45.98 ATOM 879 CB PRO 269 82.991 36.111 9.607 1.00 44.04 ATOM 880 CG PRO 269 81.631 35.506 9.4S8 1.00 43.33 ATOM 881 C PRO 269 83.710 38.492 10.004 1.00 50.31 ATOM 882 O PRO 269 84.630 38.800 10.761 1.00 49.83 ATOM 883 N ALA 270 83.48639.100 8.840 1.00 53.62 ATOM 884 CA ALA 270 84.348 40.165 8.329 1.00 54.54 ATOM 885 CB ALA 270 83.892 40.585 6.929 1.00 51.24 ATOM 886 C ALA 270 84.449 41.389 9.248 1.00 55.69 ATOM 887 O ALA 270 85.488 42.045 9.294 1.00 57.92 ATOM 888 N SER 27183.384 41.685 9.989 1.00 54.71 ATOM 889 CA SER 271 83.378 42.838 10.889 1.00 51.26 ATOM 890 CB SER 271 82.182 43.740 10.575 1.00 49.92 ATOM 891 OG SER 271 82.065 43.976 9.183 1.00 60.09 ATOM 892 C SER 271 83.305 42.443 12.360 1.00 50.78 ATOM893 O SER 271 83.482 43.288 13.240 1.00 52.11 ATOM 894 N ASP 272 83.051 41.162 12.619 1.00 48.96 ATOM 895 CA ASP 272 82.898 40.643 13.978 1.00 45.53 ATOM 896 CB ASP 272 84.206 40.765 14.776 1.00 44.82 ATOM 897 CG ASP 272 84.142 40.064 16.131 1.00 47.66 ATOM 898 OD1 ASP 272 84.750 40.581 17.091 1.00 48.64 ATOM 899 OD2 ASP 272 83.495 38.999 16.238 1.00 43.85 ATOM 900 C ASP 272 81.765 41.437 14.636 1.00 44.46 ATOM 901 O ASP 272 81.904 41.958 15.747 1.00 42.41 ATOM 902 N THR 273 80.65241.551 13.915 1.00 39.79 ATOM 903 CA THR 273 79.492 42.282 14.401 1.00 38.82 ATOM 904 CB THR 273 79.334 43.648 13.670 1.00 39.73 ATOM 905 OG1 THR 273 79.288 43.439 12.254 1.00 39.36 ATOM 906 CG2 THR 273 80.496 44.578 13.991 1.00 41.31 ATOM 907C THR 273 78.203 41.485 14.211 1.00 38.36 ATOM 908 O THR 273 78.151 40.546 13.408 1.00 33.79 ATOM 909 N LEU 274 77.187 41.835 14.995 1.00 36.91 ATOM 910 CA LEU 274 75.869 41.212 14.912 1.00 34.49 ATOM 911 CB LEU 274 75.342 40.822 16.297 1.00 30.37 ATOM 912 CG LEU 274 75.948 39.651 17.069 1.00 32.97 ATOM 913 CD1 LEU 274 75.297 39.593 18.440 1.00 28.23 ATOM 914 CD2 LEU 274 75.749 38.341 16.318 1.00 26.86 ATOM 915 C LEU 274 74.956 42.289 14.352 1.00 35.35 ATOM 916 O LEU 274 75.17143.478 14.601 1.00 37.47 ATOM 917 N THR 275 73.942 41.890 13.599 1.00 34.05 ATOM 918 CA THR 275 73.020 42.868 13.052 1.00 32.62 ATOM 919 CB THR 275 72.824 42.674 11.542 1.00 35.14 ATOM 920 OG1 THR 275 74.108 42.590 10.909 1.00 39.50 ATOM 921CG2 THR 275 72.064 43.851 10.952 1.00 30.94 ATOM 922 C THR 275 71.699 42.746 13.793 1.00 30.92 ATOM 923 O THR 275 71.100 41.670 13.845 1.00 36.53 ATOM 924 N LEU 276 71.291 43.835 14.434 1.00 28.10 ATOM 925 CA LEU 276 70.051 43.868 15.192 1.00 27.78 ATOM 926 CB LEU 276 70.205 44.780 16.420 1.00 22.51 ATOM 927 CG LEU 276 71.383 44.532 17.373 1.00 25.89 ATOM 928 CD1 LEU 276 71.225 45.408 18.608 1.00 20.70 ATOM 929 CD2 LEU 276 71.456 43.069 17.782 1.00 20.79 ATOM 930 C LEU 276 68.93044.376 14.296 1.00 27.27 ATOM 931 O LEU 276 69.068 45.430 13.672 1.00 29.06 ATOM 932 N SER 277 67.854 43.598 14.187 1.00 25.97 ATOM 933 CA SER 277 66.697 43.957 13.366 1.00 28.63 ATOM 934 CB SER 277 65.990 45.177 13.967 1.00 27.78 ATOM 935 OGSER 277 65.561 44.905 15.290 1.00 22.65 ATOM 936 C SER 277 67.067 44.209 11.897 1.00 30.31 ATOM 937 O SER 277 66.374 44.939 11.181 1.00 28.52 ATOM 938 N GLY 278 68.168 43.597 11.465 1.00 31.24 ATOM 939 CA GLY 278 68.638 43.754 10.101 1.00 39.59 ATOM 940 C GLY 278 68.999 45.178 9.706 1.00 44.55 ATOM 941 O GLY 278 69.104 45.479 8.517 1.00 46.66 ATOM 942 N GLU 279 69.234 46.046 10.686 1.00 43.47 ATOM 943 CA GLU 279 69.566 47.435 10.387 1.00 43.87 ATOM 944 CB GLU 279 68.314 48.312 10.5151.00 44.28 ATOM 945 CG GLU 279 67.703 48.322 11.908 1.00 52.30 ATOM 946 CD GLU 279 66.440 49.159 12.001 1.00 60.23 ATOM 947 OE1 GLU 279 66.398 50.074 12.853 1.00 63.06 ATOM 948 OE2 GLU 279 65.485 48.894 11.238 1.00 65.67 ATOM 949 C GLU 27970.700 48.038 11.216 1.00 42.40 ATOM 950 O GLU 279 71.330 49.001 10.787 1.00 43.89 ATOM 951 N MET 280 70.977 47.472 12.388 1.00 40.86 ATOM 952 CA MET 280 72.027 48.009 13.248 1.00 32.80 ATOM 953 CB MET 280 71.435 48.415 14.603 1.00 29.25 ATOM954 CG MET 280 72.384 49.193 15.506 1.00 31.64 ATOM 955 SD MET 280 71.830 49.235 17.232 1.00 34.02 ATOM 956 CE MET 280 70.566 50.495 17.197 1.00 26.56 ATOM 957 C MET 280 73.172 47.033 13.465 1.00 32.77 ATOM 958 O MET 280 72.983 45.971 14.058 1.D0 34.61 ATOM 959 N ALA 281 74.351 47.375 12.959 1.00 31.87 ATOM 960 CA ALA 281 75.523 46.526 13.147 1.00 34.71 ATOM 961 CB ALA 281 76.519 46.727 12.023 1.00 34.42 ATOM 962 C ALA 281 76.125 46.950 14.482 1.00 36.76 ATOM 963 O ALA 281 76.416 48.12914.693 1.00 34.59 ATOM 964 N VAL 282 76.275 45.993 15.390 1.00 37.16 ATOM 965 CA VAL 282 76.798 46.263 16.721 1.00 37.83 ATOM 966 CB VAL 282 75.692 46.023 17.780 1.00 37.58 ATOM 967 CG1 VAL 282 76.219 46.271 19.175 1.00 48.99 ATOM 968 CG2 VAL282 74.514 46.939 17.514 1.00 43.59 ATOM 969 C VAL 282 78.017 45.400 17.046 1.00 39.04 ATOM 970 O VAL 282 78.081 44.230 16.660 1.00 39.16 ATOM 971 N LYS 283 78.989 45.993 17.735 1.00 38.75 ATOM 972 CA LYS 283 80.205 45.287 18.136 1.00 42.18 ATOM 973 CB LYS 283 81.428 46.208 18.045 1.00 47.46 ATOM 974 CG LYS 283 81.803 46.617 16.632 1.00 51.71 ATOM 975 CD LYS 283 83.092 47.416 16.618 1.00 59.26 ATOM 976 CE LYS 283 83.481 47.813 15.202 1.00 62.52 ATOM 977 NZ LYS 283 82.492 48.74214.588 1.00 66.27 ATOM 978 C LYS 283 80.075 44.746 19.559 1.00 38.78 ATOM 979 O LYS 283 79.283 45.257 20.356 1.00 40.63 ATOM 980 N ARG 284 80.900 43.753 19.881 1.00 36.01 ATOM 981 CA ARG 284 80.908 43.104 21.189 1.00 38.62 ATOM 982 CB ARG 28482.150 42.224 21.327 1.00 38.83 ATOM 983 CG ARG 284 82.220 41.091 20.333 1.00 41.87 ATOM 984 CD ARG 284 83.521 40.335 20.451 1.00 39.60 ATOM 985 NE ARG 284 83.506 39.120 19.644 1.00

45.18 ATOM 986 CZ ARG 284 83.259 37.905 20.128 1.00 44.79 ATOM 987 NH1 ARG 284 83.005 37.739 21.421 1.00 41.84 ATOM 988 NH2 ARG 284 83.271 36.852 19.319 1.00 42.27 ATOM 989 C ARG 284 80.829 44.051 22.385 1.00 41.18 ATOM 990 O ARG 28479.995 43.867 23.274 1.00 44.38 ATOM 991 N GLU 285 81.703 45.052 22.416 1.00 38.71 ATOM 992 CA GLU 285 81.724 46.002 23.525 1.00 37.18 ATOM 993 CB GLU 285 82.950 46.906 23.422 1.00 36.65 ATOM 994 C GLU 285 80.444 46.838 23.614 1.00 35.71 ATOM995 O GLU 285 79.921 47.074 24.704 1.00 33.00 ATOM 996 N GLN 286 79.920 47.245 22.463 1.00 32.01 ATOM 997 CA GLN 286 78.714 48.061 22.425 1.00 32.31 ATOM 998 CB GLN 286 78.440 48.525 20.997 1.00 38.24 ATOM 999 CG GLN 286 79.565 49.352 20.392 1.00 42.42 ATOM 1000 CD GLN 286 79.277 49.761 18.964 1.00 44.79 ATOM 1001 OE1 GLN 286 79.103 48.910 18.089 1.00 42.21 ATOM 1002 NE2 GLN 286 79.215 51.063 18.719 1.00 47.53 ATOM 1003 C GLN 286 77.484 47.355 23.002 1.00 33.08 ATOM 1004 O GLN 286 76.77047.929 23.827 1.00 30.95 ATOM 1005 N LEU 287 77.245 46.114 22.579 1.00 31.49 ATOM 1006 CA LEU 287 76.095 45.350 23.068 1.00 31.01 ATOM 1007 CB LEU 287 75.892 44.073 22.242 1.00 24.63 ATOM 1008 CG LEU 287 74.498 43.780 21.661 1.00 27.34 ATOM1009 CD1 LEU 287 74.382 42.282 21.359 1.00 20.50 ATOM 1010 CD2 LEU 287 73.393 44.205 22.616 1.00 14.41 ATOM 1011 C LEU 287 76.298 44.986 24.538 1.00 32.80 ATOM 1012 O LEU 287 75.351 45.014 25.334 1.00 32.10 ATOM 1013 N LYS 288 77.536 44.64124.885 1.00 32.54 ATOM 1014 CA LYS 288 77.897 44.280 26.251 1.00 30.70 ATOM 1015 CB LYS 288 79.376 43.893 26.315 1.00 31.24 ATOM 1016 CG LYS 288 79.834 43.382 27.662 1.00 34.69 ATOM 1017 CD LYS 288 81.227 42.784 27.574 1.00 37.69 ATOM 1018 CELYS 288 81.638 42.177 28.904 1.00 42.86 ATOM 1019 NZ LYS 288 82.883 41.369 28.786 1.00 49.63 ATOM 1020 C LYS 288 77.611 45.448 27.189 1.00 28.74 ATOM 1021 O LYS 288 76.827 45.319 28.129 1.00 34.45 ATOM 1022 N ASN 289 78.190 46.602 26.882 1.00 26.57 ATOM 1023 CA ASN 289 78.011 47.803 27.691 1.00 30.84 ATOM 1024 CB ASN 289 79.012 48.819 27.274 1.00 26.04 ATOM 1025 CG ASN 289 80.437 48.485 27.570 1.00 35.16 ATOM 1026 OD1 ASN 289 80.700 47.718 28.499 1.00 42.54 ATOM 1027 ND2 ASN 28981.371 48.998 26.784 1.00 32.82 ATOM 1028 C ASN 289 76.602 48.371 27.620 1.00 35.05 ATOM 1029 O ASN 289 76.154 49.039 28.550 1.00 36.94 ATOM 1030 N GLY 290 75.909 48.113 26.515 1.00 32.43 ATOM 1031 CA GLY 290 74.556 48.614 26.345 1.00 28.66 ATOM 1032 C GLY 290 73.525 48.024 27.289 1.00 28.48 ATOM 1033 0 GLY 290 72.377 48.467 27.308 1.00 28.17 ATOM 1034 N GLY 291 73.908 47.002 28.047 1.00 28.66 ATOM 1035 CA GLY 291 72.969 46.408 28.980 1.00 29.19 ATOM 1036 C GLY 291 72.976 44.89429.075 1.00 29.76 ATOM 1037 O GLY 291 72.595 44.340 30.105 1.00 34.44 ATOM 1038 N LEU 292 73.399 44.213 28.017 1.00 29.69 ATOM 1039 CA LEU 292 73.410 42.755 28.036 1.00 30.64 ATOM 1040 CB LEU 292 73.421 42.194 26.611 1.00 27.07 ATOM 1041 CG LEU292 72.113 42.348 25.833 1.00 23.27 ATOM 1042 CD1 LEU 292 72.202 41.580 24.532 1.00 22.24 ATOM 1043 CD2 LEU 292 70.950 41.827 26.661 1.00 23.80 ATOM 1044 C LEU 292 74.530 42.125 28.861 1.00 29.22 ATOM 1045 O LEU 292 74.365 41.033 29.404 1.00 31.02 ATOM 1046 N GLY 293 75.671 42.800 28.945 1.00 30.26 ATOM 1047 CA GLY 293 76.788 42.259 29.700 1.00 28.37 ATOM 1048 C GLY 293 77.307 40.995 29.040 1.00 29.85 ATOM 1049 O GLY 293 77.460 40.951 27.820 1.00 32.37 ATOM 1050 N VAL 294 77.53739.953 29.832 1.00 30.08 ATOM 1051 CA VAL 294 78.041 38.687 29.308 1.00 31.62 ATOM 1052 CB VAL 294 78.466 37.716 30.442 1.00 29.11 ATOM 1053 CG1 VAL 294 79.649 38.292 31.191 1.00 31.37 ATOM 1054 CG2 VAL 294 77.304 37.443 31.396 1.00 26.69 ATOM1055 C VAL 294 77.079 37.978 28.351 1.00 32.81 ATOM 1056 O VAL 294 77.496 37.095 27.591 1.00 33.00 ATOM 1057 N VAL 295 75.801 38.356 28.380 1.00 30.45 ATOM 1058 CA VAL 295 74.814 37.752 27.487 1.00 28.02 ATOM 1059 CB VAL 295 73.378 38.232 27.7931.00 29.96 ATOM 1060 CG1 VAL 295 72.380 37.575 26.838 1.00 22.55 ATOM 1061 CG2 VAL 295 73.016 37.903 29.232 1.00 20.10 ATOM 1062 C VAL 295 75.203 38.115 26.057 1.00 29.90 ATOM 1063 O VAL 295 75.047 37.312 25.140 1.00 34.47 ATOM 1064 N SER 29675.762 39.309 25.886 1.00 29.11 ATOM 1065 CA SER 296 76.215 39.771 24.581 1.00 30.96 ATOM 1066 CB SER 296 76.785 41.184 24.702 1.00 27.26 ATOM 1067 OG SER 296 77.300 41.648 23.469 1.00 22.93 ATOM 1068 C SER 296 77.294 38.811 24.080 1.00 36.41 ATOM 1069 O SER 296 77.238 38.341 22.939 1.00 38.84 ATOM 1070 N ASP 297 78.254 38.501 24.954 1.00 35.29 ATOM 1071 CA ASP 297 79.346 37.585 24.629 1.00 32.14 ATOM 1072 CB ASP 297 80.245 37.356 25.851 1.00 36.57 ATOM 1073 CG ASP 297 80.958 38.61626.307 1.00 41.75 ATOM 1074 OD1 ASP 29T 81.492 39.352 25.447 1.00 45.45 ATOM 1075 OD2 ASP 297 80.999 38.861 27.532 1.00 45.15 ATOM 1076 C ASP 297 78.768 36.249 24.191 1.00 29.61 ATOM 1077 O ASP 297 79.242 35.644 23.231 1.00 32.90 ATOM 1078 NALA 298 77.738 35.804 24.903 1.00 27.85 ATOM 1079 CA ALA 298 77.071 34.544 24.608 1.00 27.89 ATOM 1080 CB ALA 298 75.998 34.258 25.657 1.00 21.67 ATOM 1081 C ALA 298 76.462 34.539 23.202 1.00 28.26 ATOM 1082 O ALA 298 76.648 33.579 22.446 1.00 30.19 ATOM 1083 N ILE 299 75.744 35.606 22.853 1.00 25.20 ATOM 1084 CA ILE 299 75.119 35.708 21.537 1.00 23.46 ATOM 1085 CB ILE 299 74.200 36.944 21.427 1.00 21.63 ATOM 1086 C 2ILE 299 73.491 36.946 20.078 1.00 22.20 ATOM 1087 CG1 ILE 299 73.14536.914 22.536 1.00 19.79 ATOM 1088 CD1 ILE 299 72.245 38.139 22.578 1.00 18.33 ATOM 1089 C ILE 299 76.181 35.752 20.444 1.00 26.28 ATOM 1090 O ILE 299 76.043 35.095 19.414 1.00 31.72 ATOM 1091 N PHE 300 77.247 36.512 20.675 1.00 29.35 ATOM 1092A PHE 300 78.338 36.613 19.709 1.00 29.01 ATOM 1093 CB PHE 300 79.386 37.622 20.182 1.00 29.53 ATOM 1094 CG PHE 300 79.239 38.978 19.562 1.00 27.60 ATOM 1095 CD1 PHE 300 78.481 39.964 20.179 1.00 24.86 ATOM 1096 CD2 PHE 300 79.853 39.266 18.3501.00 27.39 ATOM 1097 CE1 PHE 300 78.337 41.218 19.597 1.00 25.66 ATOM 1098 CE2 PHE 300 79.715 40.518 17.761 1.00 25.97 ATOM 1099 CZ PHE 300 78.956 41.495 18.384 1.00 21.03 ATOM 1100 C PHE 300 78.988 35.248 19.496 1.00 30.34 ATOM 1101 O PHE 30079.309 34.873 18.367 1.00 29.35 ATOM 1102 N GLU 301 79.181 34.507 20.582 1.00 31.04 ATOM 1103 CA GLU 301 79.775 33.178 20.499 1.00 33.60 ATOM 1104 CB GLU 301 80.012 32.607 21.898 1.00 31.64 ATOM 1105 C GLU 301 78.851 32.265 19.696 1.00 33.90 ATOM 1106 O GLU 301 79.315 31.473 18.872 1.00 33.33 ATOM 1107 N LEU 302 77.546 32.386 19.935 1.00 31.13 ATOM 1108 CA LEU 302 76.556 31.581 19.227 1.00 27.57 ATOM 1109 CB LEU 302 75.150 31.842 19.776 1.00 25.24 ATOM 1110 CG LEU 302 73.994 31.13119.059 1.00 28.59

ATOM 1111 CD1 LEU 302 74.066 29.634 19.299 1.00 25.52 ATOM 1112 CD2 LEU 302 72.660 31.682 19.532 1.00 19.30 ATOM 1113 C LEU 302 76.601 31.904 17.739 1.00 26.80 ATOM 1114 O LEU 302 76.682 31.003 16.904 1.00 27.81 ATOM 1115 N GLY 303 76.57633.195 17.416 1.00 26.47 ATOM 1116 CA GLY 303 76.611 33.624 16.030 1.00 26.99 ATOM 1117 C GLY 303 77.845 33.133 15.295 1.00 33.46 ATOM 1118 O GLY 303 77.757 32.646 14.164 1.00 32.33 ATOM 1119 N LYS 304 78.994 33.232 15.956 1.00 34.63 ATOM 1120CA LYS 304 80.269 32.813 15.383 1.00 36.20 ATOM 1121 CB LYS 304 81.399 33.115 16.372 1.00 41.96 ATOM 1122 CG LYS 304 82.779 33.179 15.757 1.00 47.05 ATOM 1123 CD LYS 304 83.800 33.610 16.796 1.00 59.47 ATOM 1124 CE LYS 304 85.179 33.791 16.1811.00 65.89 ATOM 1125 NZ LYS 304 85.182 34.863 15.144 1.00 71.01 ATOM 1126 C LYS 304 80.276 31.332 14.992 1.00 33.17 ATOM 1127 O LYS 304 80.752 30.974 13.913 1.00 34.44 ATOM 1128 N SER 305 79.739 30.482 15.861 1.00 31.40 ATOM 1129 CA SER 30579.687 29.048 15.594 1.00 33.10 ATOM 1130 CB SER 305 79.513 28.266 16.900 1.00 34.10 ATOM 1131 OG SER 305 78.391 28.727 17.633 1.00 40.61 ATOM 1132 C SER 305 78.597 28.664 14.589 1.00 33.02 ATOM 1133 O SER 305 78.771 27.718 13.816 1.00 35.32 ATOM 1134 N LEU 306 77.488 29.404 14.580 1.00 32.14 ATOM 1135 CA LEU 306 76.391 29.121 13.653 1.00 31.02 ATOM 1136 CB LEU 306 75.138 29.936 13.996 1.00 22.76 ATOM 1137 CG LEU 306 74.361 29.487 15.235 1.00 24.42 ATOM 1138 CD1 LEU 306 73.094 30.31115.380 1.00 23.13 ATOM 1139 CD2 LEU 306 74.016 28.009 15.126 1.00 25.53 ATOM 1140 C LEU 306 76.780 29.354 12.198 1.00 33.11 ATOM 1141 O LEU 306 76.161 28.796 11.293 1.00 32.60 ATOM 1142 N SER 307 77.821 30.153 11.975 1.00 36.12 ATOM 1143 CA SER307 78.296 30.448 10.624 1.00 38.80 ATOM 1144 CB SER 307 79.514 31.373 10.677 1.00 41.64 ATOM 1145 OG SER 307 79.224 32.556 11.401 1.00 54.66 ATOM 1146 C SER 307 78.650 29.182 9.845 1.00 36.98 ATOM 1147 O SER 307 78.302 29.055 8.669 1.00 42.87 ATOM 1148 N ALA 308 79.315 28.239 10.509 1.00 35.72 ATOM 1149 CA ALA 308 79.719 26.983 9.879 1.00 32.70 ATOM 1150 CB ALA 308 80.683 26.227 10.782 1.00 33.88 ATOM 1151 C ALN 308 78.531 26.093 9.521 1.00 34.83 ATOM 1152 O ALA 308 78.620 25.2788.600 1.00 39.61 ATOM 1153 N PHE 309 77.424 26.250 10.244 1.00 31.54 ATOM 1154 CA PHE 309 76.226 25.453 9.999 1.00 32.43 ATOM 1155 CB PHE 309 75.259 25.558 11.182 1.00 30.89 ATOM 1156 CG PHE 309 75.718 24.826 12.415 1.00 33.73 ATOM 1157 CD1 PHE309 76.769 25.314 13.183 1.00 40.48 ATOM 1158 CD2 PHE 309 75.091 23.654 12.816 1.00 35.96 ATOM 1159 CE1 PHE 309 77.189 24.643 14.334 1.00 37.87 ATOM 1160 CE2 PHE 309 75.502 22.975 13.962 1.00 38.44 ATOM 1161 CZ PHE 309 76.553 23.471 14.722 1.00 37.34 ATOM 1162 C PHE 309 75.507 25.809 8.693 1.00 34.76 ATOM 1163 O PHE 309 74.810 24.969 8.118 1.00 36.18 ATOM 1164 N ASN 310 75.693 27.040 8.218 1.00 35.80 ATOM 1165 CA ASN 310 75.060 27.506 6.980 1.00 41.00 ATOM 1166 CB ASN 310 75.705 26.8525.755 1.00 51.94 ATOM 1167 CG ASN 310 77.053 27.452 5.419 1.00 67.92 ATOM 1168 OD1 ASN 310 77.139 28.439 4.687 1.00 77.32 ATOM 1169 ND2 ASN 310 78.116 26.869 5.962 1.00 72.62 ATOM 1170 C ASN 310 73.560 27.245 6.985 1.00 38.15 ATOM 1171 O ASN310 73.034 26.515 6.141 1.00 35.87 ATOM 1172 N LEU 311 72.885 27.819 7.971 1.00 33.94 ATOM 1173 CA LEU 311 71.450 27.651 8.111 1.00 32.09 ATOM 1174 CB LEU 311 71.011 28.009 9.533 1.00 28.06 ATOM 1175 CG LEU 311 71.656 27.301 10.724 1.00 26.38 ATOM 1176 CD1 LEU 311 71.092 27.883 12.012 1.00 23.56 ATOM 1177 CD2 LEU 311 71.409 25.801 10.651 1.00 21.24 ATOM 1178 C LEU 311 70.705 28.542 7.124 1.00 33.00 ATOM 1179 O LEU 311 71.173 29.630 6.782 1.00 35.47 ATOM 1180 N ASP 312 69.569 28.0576.638 1.00 27.78 ATOM 1181 CA ASP 312 68.749 28.841 5.733 1.00 27.06 ATOM 1182 CB ASP 312 68.385 28.049 4.456 1.00 25.84 ATOM 1183 CG ASP 312 67.580 26.778 4.724 1.00 25.67 ATOM 1184 OD1 ASP 312 67.124 26.541 5.860 1.00 28.20 ATOM 1185 OD2 ASP312 67.387 26.008 3.762 1.00 27.62 ATOM 1186 C ASP 312 67.517 29.314 6.514 1.00 28.51 ATOM 1187 O ASP 312 67.371 28.990 7.703 1.00 25.35 ATOM 1188 N ASP 313 66.633 30.060 5.855 1.00 22.16 ATOM 1189 CA ASP 313 65.430 30.589 6.494 1.00 21.37 ATOM1190 CB ASP 313 64.625 31.431 5.499 1.00 25.11 ATOM 1191 CG ASP 313 65.380 32.666 5.025 1.00 31.54 ATOM 1192 OD1 ASP 313 65.119 33.115 3.890 1.00 35.35 ATOM 1193 OD2 ASP 313 66.225 33.193 5.783 1.00 35.37 ATOM 1194 C ASP 313 64.524 29.535 7.1201.00 21.11 ATOM 1195 O ASP 313 63.904 29.783 8.158 1.00 23.68 ATOM 1196 N THR 314 64.440 28.367 6.489 1.00 22.88 ATOM 1197 CA THR 314 63.591 27.281 6.981 1.00 22.81 ATOM 1198 CB THR 314 63.472 26.155 5.927 1.00 26.00 ATOM 1199 OG1 THR 31462.873 26.679 4.732 1.00 20.14 ATOM 1200 CG2 THR 314 62.629 25.010 6.457 1.00 17.51 ATOM 1201 C THR 314 64.086 26.706 8.310 1.00 19.46 ATOM 1202 O THR 314 63.312 26.529 9.247 1.00 19.33 ATOM 1203 N GLU 315 65.381 26.431 8.392 1.00 17.49 ATOM1204 CA GLU 315 65.965 25.885 9.611 1.00 20.62 ATOM 1205 CB GLU 315 67.426 25.514 9.358 1.00 14.39 ATOM 1206 CG GLU 315 67.539 24.339 8.400 1.00 13.07 ATOM 1207 CD GLU 315 68.923 24.125 7.835 1.00 14.98 ATOM 1208 OE1 GLU 315 69.634 25.116 7.5521.00 17.71 ATOM 1209 OE2 GLU 315 69.287 22.948 7.651 1.00 17.88 ATOM 1210 C GLU 315 65.810 26.883 10.762 1.00 20.57 ATOM 1211 O GLU 315 65.368 26.518 11.854 1.00 18.43 ATOM 1212 N VAL 316 66.096 28.154 10.488 1.00 19.19 ATOM 1213 CA VAL 31665.955 29.203 11.490 1.00 16.53 ATOM 1214 CB VAL 316 66.418 30.567 10.933 1.00 17.42 ATOM 1215 CG1 VAL 316 66.149 31.687 11.940 1.00 13.89 ATOM 1216 CG2 VAL 316 67.900 30.506 10.594 1.00 14.31 ATOM 1217 C VAL 316 64.488 29.291 11.927 1.00 19.53 ATOM 1218 O VAL 316 64.191 29.448 13.110 1.00 19.86 ATOM 1219 N ALA 317 63.575 29.159 10.970 1.00 19.02 ATOM 1220 CA ALA 317 62.145 29.215 11.254 1.00 16.95 ATOM 1221 CB ALA 317 61.357 29.239 9.951 1.00 17.68 ATOM 1222 C ALA 317 61.674 28.04712.126 1.00 14.13 ATOM 1223 O ALA 317 60.875 28.228 13.045 1.00 15.34 ATOM 1224 N LEU 318 62.154 26.847 11.819 1.00 17.41 ATOM 1225 CA LEU 318 61.769 25.653 12.569 1.00 19.10 ATOM 1226 CB LEU 318 62.186 24.398 11.802 1.00 18.21 ATOM 1227 CG LEU318 61.443 24.209 10.473 1.00 19.02 ATOM 1228 CD1 LEU 318 62.105 23.128 9.646 1.00 16.10 ATOM 1229 CD2 LEU 318 59.987 23.875 10.735 1.00 11.32 ATOM 1230 C LEU 318 62.399 25.685 13.954 1.00 22.38 ATOM 1231 O LEU 318 61.782 25.278 14.945 1.00 21.64 ATOM 1232 N LEU 319 63.619 26.207 14.016 1.00 20.97 ATOM 1233 CA LEU 319 64.338 26.344 15.270 1.00 19.71 ATOM 1234 CB LEU 319 65.715 26.951 15.005 1.00 20.56 ATOM 1235 CG LEU 319 66.722 27.036 16.152 1.00 32.05 ATOM 1236 CD1 LEU 319 66.70425.760 16.963 1.00

33.15 ATOM 1237 CD2 LEU 319 68.109 27.303 15.590 1.00 28.25 ATOM 1238 C LEU 319 63.496 27.254 16.164 1.00 20.66 ATOM 1239 O LEU 319 63.215 26.920 17.313 1.00 24.47 ATOM 1240 N GLN 320 63.026 28.365 15.604 1.00 19.25 ATOM 1241 CA GLN 32062.191 29.307 16.346 1.00 19.02 ATOM 1242 CB GLN 320 61.842 30.526 15.488 1.00 19.11 ATOM 1243 CG GLN 320 63.032 31.377 15.101 1.00 20.02 ATOM 1244 CD GLN 320 62.665 32.562 14.224 1.00 23.65 ATOM 1245 OE1 GLN 320 63.487 33.445 13.997 1.00 22.68 ATOM 1246 NE2 GLN 320 61.440 32.574 13.704 1.00 20.77 ATOM 1247 C GLN 320 60.905 28.635 16.811 1.00 20.52 ATOM 1248 O GLN 320 60.465 28.845 17.938 1.00 22.04 ATOM 1249 N ALA 321 60.306 27.825 15.942 1.00 21.01 ATOM 1250 CA ALA 321 59.069 27.12816.280 1.00 16.83 ATOM 1251 CB ALA 321 58.556 26.358 15.079 1.00 16.58 ATOM 1252 C ALA 321 59.288 26.185 17.462 1.00 18.15 ATOM 1253 O ALA 321 58.427 26.069 18.344 1.00 13.03 ATOM 1254 N VAL 322 60.442 25.523 17.481 1.00 14.89 ATOM 1255 CA VAL322 60.774 24.599 18.559 1.00 19.05 ATOM 1256 CB VAL 322 62.051 23.779 18.233 1.00 21.50 ATOM 1257 CG1 VAL 322 62.510 22.990 19.457 1.00 21.49 ATOM 1258 CG2 VAL 322 61.773 22.819 17.073 1.00 15.42 ATOM 1259 C VAL 322 60.947 25.375 19.867 1.00 19.89 ATOM 1260 O VAL 322 60.478 24.940 20.919 1.00 21.58 ATOM 1261 N LEU 323 61.591 26.537 19.788 1.00 20.25 ATOM 1262 CA LEU 323 61.804 27.387 20.959 1.00 19.32 ATOM 1263 CB LEU 323 62.683 28.586 20.597 1.00 12.95 ATOM 1264 CG LEU 323 64.12928.273 20.217 1.00 20.70 ATOM 1265 CD1 LEU 323 64.805 29.503 19.641 1.00 13.23 ATOM 1266 CD2 LEU 323 64.883 27.767 21.438 1.00 22.91 ATOM 1267 C LEU 323 60.468 27.884 21.497 1.00 20.25 ATOM 1268 O LEU 323 60.251 27.918 22.706 1.00 25.88 ATOM1269 N LEU 324 59.571 28.251 20.587 1.00 23.08 ATOM 1270 CA LEU 324 58.248 28.753 20.944 1.00 21.24 ATOM 1271 CB LEU 324 57.555 29.333 19.707 1.00 18.45 ATOM 1272 CG LEU 324 56.119 29.847 19.868 1.00 17.07 ATOM 1273 CD1 LEU 324 56.083 31.09220.752 1.00 15.39 ATOM 1274 CD2 LEU 324 55.545 30.162 18.498 1.06 17.90 ATOM 1275 C LEU 324 57.342 27.706 21.598 1.00 21.54 ATOM 1276 O LEU 324 56.742 27.967 22.642 1.00 23.41 ATOM 1277 N MET 325 57.249 26.521 21.003 1.00 24.63 ATOM 1278 CA MET325 56.380 25.476 21.545 1.00 25.35 ATOM 1279 CB MET 325 55.901 24.536 20.430 1.00 25.53 ATOM 1280 CG MET 325 55.235 25.220 19.232 1.00 21.89 ATOM 1281 SD MET 325 53.871 26.337 19.649 1.00 25.50 ATOM 1282 CE MET 325 52.705 25.250 20.397 1.00 17.66 ATOM 1283 C MET 325 57.031 24.676 22.675 1.00 27.58 ATOM 1284 O MET 325 56.988 23.450 22.690 1.00 28.61 ATOM 1285 N SER 326 57.613 25.376 23.638 1.00 27.98 ATOM 1286 CA SER 326 58.265 24.718 24.757 1.00 31.60 ATOM 1287 CB SER 326 59.52725.493 25.155 1.00 35.80 ATOM 1288 OG SER 326 60.123 24.966 26.327 1.00 43.74 ATOM 1289 C SER 326 57.313 24.624 25.939 1.00 32.12 ATOM 1290 O SER 326 56.590 25.574 26.240 1.00 30.94 ATOM 1291 N THR 327 57.276 23.464 26.583 1.00 35.41 ATOM 1292CA THR 327 56.420 23.278 27.747 1.00 39.61 ATOM 1293 CB THR 327 55.777 21.890 27.758 1.00 38.84 ATOM 1294 OG1 THR 327 56.784 20.890 27.538 1.00 42.53 ATOM 1295 CG2 THR 327 54.716 21.802 26.679 1.00 40.78 ATOM 1296 C THR 327 57.232 23.471 29.0221.00 43.86 ATOM 1297 O THR 327 56.785 23.133 30.118 1.00 42.40 ATOM 1298 N ASP 328 58.417 24.054 28.869 1.00 47.35 ATOM 1299 CA ASP 328 59.309 24.308 29.987 1.00 49.43 ATOM 1300 CB ASP 328 60.750 24.358 29.482 1.00 58.03 ATOM 1301 CG ASP 32861.718 23.687 30.425 1.00 72.16 ATOM 1302 OD1 ASP 328 61.816 24.117 31.595 1.00 82.32 ATOM 1303 OD2 ASP 328 62.378 22.720 29.994 1.00 81.63 ATOM 1304 C ASP 328 58.951 25.625 30.676 1.00 47.99 ATOM 1305 O ASP 328 59.830 26.373 31.093 1.00 53.33 ATOM 1306 N ARG 329 57.657 25.910 30.780 1.00 48.33 ATOM 1307 CA ARG 329 57.177 27.135 31.413 1.00 47.67 ATOM 1308 CB ARG 329 56.562 28.091 30.379 1.00 47.64 ATOM 1309 CG ARG 329 57.550 28.802 29.450 1.00 47.87 ATOM 1310 CD ARG 329 57.893 27.96828.226 1.00 44.00 ATOM 1311 NE ARG 329 58.759 28.682 27.288 1.00 41.17 ATOM 1312 CZ ARG 329 60.087 28.605 27.283 1.00 48.58 ATOM 1313 NH1 ARG 329 60.719 27.848 28.172 1.00 52.94 ATOM 1314 NH2 ARG 329 60.784 29.257 26.362 1.00 43.16 ATOM 1315 CARG 329 56.126 26.778 32.457 1.00 48.01 ATOM 1316 O ARG 329 55.573 25.677 32.437 1.00 50.22 ATOM 1317 N SER 330 55.832 27.716 33.351 1.00 47.37 ATOM 1318 CA SER 330 54.848 27.490 34.402 1.00 47.64 ATOM 1319 CB SER 330 55.376 28.021 35.736 1.00 46.62 ATOM 1320 C SER 330 53.506 28.139 34.074 1.00 46.40 ATOM 1321 O SER 330 53.460 29.252 33.548 1.00 48.49 ATOM 1322 N GLY 331 52.421 27.424 34.359 1.00 44.16 ATOM 1323 CA GLY 331 51.090 27.956 34.123 1.00 41.44 ATOM 1324 C GLY 331 50.42427.660 32.790 1.00 42.83 ATOM 1325 O GLY 331 49.478 28.351 32.413 1.00 45.88 ATOM 1326 N LEU 332 50.889 26.643 32.075 1.00 40.10 ATOM 1327 CA LEU 332 50.288 26.300 30.789 1.00 39.27 ATOM 1328 CB LEU 332 51.301 25.596 29.885 1.00 37.42 ATOM 1329CG LEU 332 52.436 26.426 29.291 1.00 35.35 ATOM 1330 CD1 LEU 332 53.374 25.505 28.530 1.00 31.61 ATOM 1331 CD2 LEU 332 51.875 27.511 28.376 1.00 31.82 ATOM 1332 C LEU 332 49.058 25.415 30.951 1.00 39.32 ATOM 1333 O LEU 332 49.060 24.467 31.7381.00 42.74 ATOM 1334 N LEU 333 48.009 25.730 30.202 1.00 37.62 ATOM 1335 CA LEU 333 46.778 24.953 30.241 1.00 41.30 ATOM 1336 CB LEU 333 45.586 25.835 29.852 1.00 43.52 ATOM 1337 CG LEU 333 45.125 26.904 30.848 1.00 49.39 ATOM 1338 CD1 LEU 33344.296 27.970 30.142 1.00 46.19 ATOM 1339 CD2 LEU 333 44.330 26.248 31.968 1.00 51.29 ATOM 1340 C LEU 333 46.859 23.762 29.285 1.00 41.39 ATOM 1341 O LEU 333 46.565 22.628 29.657 1.00 43.41 ATOM 1342 N CYA 334 47.3.17 24.024 28.067 1.00 42.18 ATOM 1343 CA CYA 334 47.409 23.003 27.029 1.00 39.56 ATOM 1344 CB CYA 334 47.004 23.616 25.691 1.00 45.48 ATOM 1345 SG CYA 334 45.517 24.616 25.785 1.00 51.57 ATOM 1346 AS CYA 334 44.187 22.808 25.555 1.00 90.90 ATOM 1347 C CYA 334 48.776 22.34726.891 1.00 38.28 ATOM 1348 O CYA 334 49.273 22.178 25.778 1.00 40.95 ATOM 1349 N VAL 335 49.345 21.913 28.009 1.00 36.05 ATOM 1350 CA VAL 335 50.661 21.278 28.006 1.00 35.78 ATOM 1351 CB VAL 335 50.996 20.679 29.399 1.00 35.53 ATOM 1352 CG1VAL 335 52.413 20.123 29.407 1.00 32.76 ATOM 1353 CG2 VAL 335 50.822 21.729 30.490 1.00 28.87 ATOM 1354 C VAL 335 50.776 20.170 26.950 1.00 36.41 ATOM 1355 O VAL 335 51.756 20.104 26.202 1.00 34.26 ATOM 1356 N ASP 336 49.756 19.323 26.880 1.00 38.42 ATOM 1357 CA ASP 336 49.736 18.209 25.942 1.00 39.71 ATOM 1358 CB ASP 336 48.485 17.359 26.179 1.00 51.53 ATOM 1359 CG ASP 336 48.534 16.028 25.452 1.00 65.98 ATOM 1360 OD1 ASP 336 49.240 15.114 25.934 1.00 70.75 ATOM 1361 OD2 ASP 33647.858 15.891 24.406 1.00 72.15

ATOM 1362 C ASP 336 49.794 18.668 24.486 1.00 37.72 ATOM 1363 O ASP 336 50.686 18.259 23.733 1.00 32.08 ATOM 1364 N LYS 337 48.858 19.532 24.100 1.00 33.78 ATOM 1365 CA LYS 337 48.797 20.040 22.731 1.00 28.00 ATOM 1366 CB LYS 337 47.62621.022 22.574 1.00 22.46 ATOM 1367 C LYS 337 50.116 20.704 22.334 1.00 29.06 ATOM 1368 O LYS 337 50.607 20.512 21.220 1.00 28.41 ATOM 1369 N ILE 338 50.705 21.449 23.267 1.00 27.56 ATOM 1370 CA ILE 338 51.964 22.138 23.022 1.00 25.03 ATOM 1371CB ILE 338 52.274 23.149 24.144 1.00 19.49 ATOM 1372 CG2 ILE 338 53.577 23.876 23.859 1.00 19.00 ATOM 1373 CG1 ILE 338 51.135 24.167 24.232 1.00 21.97 ATOM 1374 CD1 ILE 338 51.277 25.175 25.348 1.00 26.67 ATOM 1375 C ILE 338 53.119 21.153 22.8261.00 29.97 ATOM 1376 O ILE 338 53.935 21.328 21.914 1.00 31.00 ATOM 1377 N GLU 339 53.165 20.100 23.642 1.00 33.52 ATOM 1378 CA GLU 339 54.213 19.080 23.516 1.00 35.34 ATOM 1379 CB GLU 339 54.136 18.062 24.659 1.00 39.97 ATOM 1380 CG GLU 33954.653 18.585 25.986 1.00 53.23 ATOM 1381 CD GLU 339 54.549 17.579 27.126 1.00 61.16 ATOM 1382 OE1 GLU 339 53.602 16.759 27.131 1.00 64.30 ATOM 1383 OE2 GLU 339 55.412 17.622 28.031 1.00 57.76 ATOM 1384 C GLU 339 54.091 18.353 22.178 1.00 31.63 ATOM 1385 O GLU 339 55.086 18.123 21.491 1.00 28.96 ATOM 1386 N LYS 340 52.861 18.006 21.810 1.00 30.95 ATOM 1387 CA LYS 340 52.602 17.313 20.554 1.00 31.58 ATOM 1388 CB LYS 340 51.121 16.966 20.438 1.00 31.83 ATOM 1389 C LYS 340 53.057 18.15919.358 1.00 29.84 ATOM 1390 O LYS 340 53.696 17.640 18.438 1.00 31.58 ATOM 1391 N SER 341 52.765 19.460 19.388 1.00 25.33 ATOM 1392 CA SER 341 53.165 20.351 18.297 1.00 23.92 ATOM 1393 CB SER 341 52.468 21.707 18.400 1.00 24.02 ATOM 1394 OG SER341 52.700 22.302 19.657 1.00 48.88 ATOM 1395 C SER 341 54.677 20.533 18.240 1.00 24.39 ATOM 1396 O SER 341 55.254 20.593 17.150 1.00 24.71 ATOM 1397 N GLN 342 55.324 20.606 19.405 1.00 25.45 ATOM 1398 CA GLN 342 56.777 20.751 19.437 1.00 26.66 ATOM 1399 CB GLN 342 57.311 20.975 20.853 1.00 22.77 ATOM 1400 CG GLN 342 58.805 21.307 20.840 1.00 25.76 ATOM 1401 CD GLN 342 59.427 21.371 22.214 1.00 28.46 ATOM 1402 OE1 GLN 342 59.342 20.422 22.990 1.00 34.22 ATOM 1403 NE2 GLN 342 60.08022.483 22.517 1.00 30.01 ATOM 1404 C GLN 342 57.425 19.504 18.843 1.00 23.37 ATOM 1405 O GLN 342 58.414 19.598 18.106 1.00 23.65 ATOM 1406 N GLU 343 56.864 18.340 19.162 1.00 21.48 ATOM 1407 CA GLU 343 57.370 17.076 18.641 1.00 20.74 ATOM 1408CB GLU 343 56.599 15.902 19.247 1.00 22.09 ATOM 1409 C GLU 343 57.225 17.094 17.119 1.00 19.18 ATOM 1410 O GLU 343 58.156 16.743 16.393 1.00 21.11 ATOM 1411 N ALA 344 56.077 17.570 16.648 1.00 19.93 ATOM 1412 CA ALA 344 55.803 17.662 15.217 1.00 20.20 ATOM 1413 CB ALA 344 54.411 18.216 14.989 1.00 16.46 ATOM 1414 C ALA 344 56.850 18.539 14.528 1.00 20.75 ATOM 1415 O ALA 344 57.432 18.140 13.514 1.00 25.13 ATOM 1416 N TYR 345 57.105 19.722 15.088 1.00 21.31 ATOM 1417 CA TYR 345 58.10720.631 14.531 1.00 15.93 ATOM 1418 CB TYR 345 58.127 21.969 15.282 1.00 17.29 ATOM 1419 CG TYR 345 57.049 22.927 14.833 1.00 16.11 ATOM 1420 CD1 TYR 345 56.017 23.296 15.689 1.00 9.93 ATOM 1421 CE1 TYR 345 54.999 24.138 15.263 1.00 16.95 ATOM1422 CD2 TYR 345 57.041 23.431 13.531 1.00 19.84 ATOM 1423 CE2 TYR 345 56.026 24.276 13.094 1.00 17.13 ATOM 1424 CZ TYR 345 5S.005 24.622 13.963 1.00 18.12 ATOM 1425 OH TYR 345 53.980 25.430 13.530 1.00 26.25 ATOM 1426 C TYR 345 59.493 20.00814.554 1.00 20.65 ATOM 1427 O TYR 345 60.240 20.129 13.583 1.00 20.75 ATOM 1428 N LEU 346 59.832 19.337 15.655 1.00 22.14 ATOM 1429 CA LEU 346 61.134 18.684 15.803 1.00 19.43 ATOM 1430 CB LEU 346 61.267 18.041 17.186 1.00 19.92 ATOM 1431 CG LEU346 61.683 18.945 18.347 1.00 25.56 ATOM 1432 CD1 LEU 346 61.440 18.244 19.677 1.00 22.06 ATOM 1433 CD2 LEU 346 63.147 19.332 18.197 1.00 17.62 ATOM 1434 C LEU 346 61.359 17.635 14.723 1.00 19.30 ATOM 1435 O LEU 346 62.441 17.560 14.142 1.00 22.84 ATOM 1436 N LEU 347 60.337 16.826 14.456 1.00 25.17 ATOM 1437 CA LEU 347 60.423 15.790 13.427 1.00 24.55 ATOM 1438 CB LEU 347 59.187 14.892 13.453 1.00 25.47 ATOM 1439 CG LEU 347 59.2S6 13.654 14.345 1.00 30.65 ATOM 1440 CD1 LEU 347 57.94112.890 14.258 1.00 34.28 ATOM 1441 CD2 LEU 347 60.416 12.765 13.908 1.00 28.26 ATOM 1442 C LEU 347 60.584 16.400 12.042 1.00 24.00 ATOM 1443 O LEU 347 61.399 15.932 11.245 1.00 29.74 ATOM 1444 N ALA 348 59.809 17.443 11.761 1.00 22.72 ATOM 1445CA ALA 348 59.875 18.125 10.475 1.00 19.19 ATOM 1446 CB ALA 348 58.789 19.188 10.388 1.00 22.73 ATOM 1447 C ALA 348 61.246 18.762 10.316 1.00 20.34 ATOM 1448 O ALA 348 61.881 18.633 9.274 1.00 23.94 ATOM 1449 N PHE 349 61.707 19.402 11.388 1.00 22.19 ATOM 1450 CA PHE 349 63.001 20.078 11.435 1.00 19.41 ATOM 1451 CB PHE 349 63.185 20.701 12.832 1.00 17.45 ATOM 1452 CG PHE 349 64.371 21.632 12.963 1.00 18.70 ATOM 1453 CD1 PHE 349 65.183 21.943 11.874 1.00 19.09 ATOM 1454 CD2 PHE 34964.669 22.203 14.199 1.00 21.81 ATOM 1455 CE1 PHE 349 66.270 22.811 12.012 1.00 21.49 ATOM 1456 CE2 PHE 349 65.753 23.072 14.351 1.00 18.58 ATOM 1457 CZ PHE 349 66.555 23.376 13.256 1.00 18.67 ATOM 1458 C PHE 349 64.110 19.071 11.136 1.00 20.96 ATOM 1459 O PHE 349 64.961 19.311 10.283 1.00 25.19 ATOM 1460 N GLU 350 64.076 17.935 11.824 1.00 23.96 ATOM 1461 CA GLU 350 65.077 16.888 11.642 1.00 27.98 ATOM 1462 CB GLU 350 64.794 15.721 12.591 1.00 28.90 ATOM 1463 CG GLU 350 65.738 14.54212.413 1.00 39.36 ATOM 1464 CD GLU 350 65.603 13.497 13.505 1.00 41.62 ATOM 1465 OE1 GLU 350 64.475 13.260 13.988 1.00 43.67 ATOM 1466 OE2 GLU 350 66.636 12.908 13.876 1.00 49.64 ATOM 1467 C GLU 350 65.100 16.385 10.203 1.00 27.12 ATOM 1468 OGLU 350 66.158 16.288 9.577 1.00 27.44 ATOM 1469 N HIS 351 63.918 16.088 9.678 1.00 27.36 ATOM 1470 CA HIS 351 63.787 15.591 8.318 1.00 23.97 ATOM 1471 CB HIS 351 62.366 15.087 8.090 1.00 22.89 ATOM 1472 CG HIS 351 61.991 13.945 8.986 1.00 24.58 ATOM 1473 CD2 HIS 351 62.736 13.209 9.846 1.00 25.83 ATOM 1474 ND1 HIS 351 60.709 13.448 9.073 1.00 26.50 ATOM 1475 CE1 HIS 351 60.677 12.460 9.948 1.00 24.81 ATOM 1476 NE2 HIS 351 61.896 12.295 10.431 1.00 28.42 ATOM 1477 C HIS 351 64.200 16.6357.278 1.00 24.22 ATOM 1478 O HIS 351 64.757 16.287 6.236 1.00 25.79 ATOM 1479 N TYR 352 63.969 17.912 7.572 1.00 21.04 ATOM 1480 CA TYR 352 64.363 18.974 6.654 1.00 18.98 ATOM 1481 CB TYR 352 63.770 20.321 7.067 1.00 17.08 ATOM 1482 CG TYR 35264.127 21.413 6.090 1.00 21.83 ATOM 1483 CD1 TYR 352 63.537 21.467 4.828 1.00 20.07 ATOM 1484 CE1 TYR 352 63.941 22.411 3.883 1.00 23.51 ATOM 1485 CD2 TYR 352 65.121 22.339 6.388 1.00 19.94 ATOM 1486 CE2 TYR 352 65.531 23.284 5.452 1.00 20.85 ATOM 1487 CZ TYR 352 64.942 23.313 4.203 1.00

24.80 ATOM 1488 OH TYR 352 65.380 24.221 3.269 1.00 26.74 ATOM 1489 C TYR 352 65.889 19.055 6.624 1.00 20.58 ATOM 1490 O TYR 352 66.492 19.276 5.570 1.00 22.72 ATOM 1491 N VAL 353 66.508 18.877 7.789 1.00 28.34 ATOM 1492 CA VAL 35367.967 18.892 7.904 1.00 22.38 ATOM 1493 CB VAL 353 68.419 18.755 9.389 1.00 26.46 ATOM 1494 CG1 VAL 353 69.915 18.527 9.478 1.00 20.92 ATOM 1495 CG2 VAL 353 68.053 20.009 10.165 1.00 22.46 ATOM 1496 C VAL 353 68.518 17.725 7.078 1.00 23.51 ATOM 1497 O VAL 353 69.535 17.865 6.391 1.00 24.73 ATOM 1498 N ASN 354 67.850 16.575 7.158 1.00 20.93 ATOM 1499 CA ASN 354 68.252 15.392 6.397 1.00 27.25 ATOM 1500 CB ASN 354 67.320 14.210 6.680 1.00 28.43 ATOM 1501 CG ASN 354 67.521 13.607 8.0581.00 31.50 ATOM 1502 OD1 ASN 354 68.565 13.787 8.692 1.00 37.79 ATOM 1503 ND2 ASN 354 66.521 12.867 8.524 1.00 26.44 ATOM 1504 C ASN 354 68.182 15.721 4.908 1.00 31.27 ATOM 1505 O ASN 354 69.066 15.347 4.134 1.00 34.22 ATOM 1506 N HIS 35567.124 16.429 4.520 1.00 30.49 ATOM 1507 CA HIS 355 66.917 16.826 3.132 1.00 26.88 ATOM 1508 CB HIS 355 65.548 17.494 2.975 1.00 27.27 ATOM 1509 CG HIS 355 65.319 18.103 1.625 1.00 37.76 ATOM 1510 CD2 HIS 355 65.439 19.382 1.196 1.00 35.28 ATOM1511 ND1 HIS 355 64.913 17.369 0.532 1.00 34.93 ATOM 1512 CE1 HIS 355 64.789 18.169 -0.513 1.00 34.84 ATOM 1513 NE2 HIS 355 65.104 19.394 -0.135 1.00 33.13 ATOM 1514 C HIS 355 68.016 17.748 2.610 1.00 24.66 ATOM 1515 O HIS 355 68.420 17.630 1.4561.00 26.62 ATOM 1516 N ARG 356 68.487 18.670 3.448 1.00 25.86 ATOM 1517 CA ARG 356 69.536 19.608 3.040 1.00 26.94 ATOM 1518 CB ARG 356 69.620 20.791 3.996 1.00 20.57 ATOM 1519 CG ARG 356 68.453 21.727 3.899 1.00 19.69 ATOM 1520 CD ARG 35668.866 23.110 4.340 1.00 23.81 ATOM 1521 NE ARG 356 69.768 23.746 3.388 1.00 23.14 ATOM 1522 CZ ARG 356 70.641 24.697 3.702 1.00 24.11 ATOM 1523 NH1 ARG 356 70.755 25.129 4.949 1.00 26.29 ATOM 1524 NH2 ARG 356 71.384 25.242 2.754 1.00 32.79 ATOM 1525 C ARG 356 70.921 19.002 2.875 1.00 29.38 ATOM 1526 O ARG 356 71.795 19.607 2.257 1.00 32.91 ATOM 1527 N LYS 357 71.133 17.848 3.498 1.00 33.39 ATOM 1528 CA LYS 357 72.401 17.128 3.417 1.00 35.97 ATOM 1529 CB LYS 357 72.479 16.363 2.0891.00 40.55 ATOM 1530 CG LYS 357 71.327 15.381 1.891 1.00 44.03 ATOM 1531 CD LYS 357 71.360 14.722 0.523 1.00 52.31 ATOM 1532 CE LYS 357 70.171 13.787 0.343 1.00 56.99 ATOM 1533 NZ LYS 357 70.208 13.085 -0.970 1.00 64.78 ATOM 1534 C LYS 35773.657 17.981 3.629 1.00 38.55 ATOM 1535 O LYS 357 74.518 18.079 2.748 1.00 42.50 ATOM 1536 N HIS 358 73.751 18.601 4.802 1.00 35.00 ATOM 1537 CA HIS 358 74.906 19.418 5.155 1.00 32.94 ATOM 1538 CB HIS 358 74.732 20.018 6.552 1.00 27.62 ATOM1539 CG HIS 358 73.669 21.067 6.643 1.00 26.64 ATOM 1540 CD2 HIS 358 72.330 20.968 6.819 1.00 20.85 ATOM 1541 ND1 HIS 358 73.950 22.416 6.587 1.00 24.71 ATOM 1542 CE1 HIS 358 72.831 23.103 6.724 1.00 21.02 ATOM 1543 NE2 HIS 358 71.834 22.2486.865 1.00 21.42 ATOM 1544 C HIS 358 76.140 18.520 5.176 1.00 36.60 ATOM 1545 O HIS 358 76.072 17.379 5.635 1.00 38.73 ATOM 1546 N ASN 359 77.267 19.037 4.702 1.00 41.40 ATOM 1547 CA ASN 359 78.515 18.277 4.689 1.00 45.02 ATOM 1548 CB ASN 35979.441 18.799 3.587 1.00 42.57 ATOM 1549 C ASN 359 79.193 18.386 6.058 1.00 46.59 ATOM 1550 O ASN 359 80.405 18.588 6.150 1.00 52.31 ATOM 1551 N ILE 360 78.400 18.254 7.117 1.00 45.14 ATOM 1552 CA ILE 360 78.896 18.348 8.487 1.00 43.69 ATOM1553 CB ILE 360 78.330 19.597 9.207 1.00 40.08 ATOM 1554 CG2 ILE 360 78.824 19.657 10.645 1.00 32.11 ATOM 1555 CG1 ILE 360 78.733 20.864 8.452 1.00 41.47 ATOM 1556 CD1 ILE 360 78.057 22.115 8.954 1.00 44.93 ATOM 1557 C ILE 360 78.452 17.101 9.2421.00 43.63 ATOM 1558 O ILE 360 77.257 16.797 9.313 1.00 45.20 ATOM 1559 N PRO 361 79.413 16.337 9.780 1.00 43.91 ATOM 1560 CD PRO 361 80.871 16.540 9.699 1.00 47.07 ATOM 1561 CA PRO 361 79.087 15.118 10.526 1.00 41.66 ATOM 1562 CB PRO 36180.462 14.495 10.782 1.00 43.73 ATOM 1563 CG PRO 361 81.383 15.679 10.830 1.00 45.45 ATOM 1564 C PRO 361 78.332 15.403 11.832 1.00 36.42 ATOM 1565 O PRO 361 78.679 16.325 12.572 1.00 35.74 ATOM 1566 N HIS 362 77.291 14.610 12.088 1.00 33.14 ATOM 1567 CA HIS 362 76.462 14.726 13.292 1.00 34.09 ATOM 1568 CB HIS 362 77.288 14.413 14.547 1.00 33.82 ATOM 1569 CG HIS 362 78.132 13.18 14.424 1.00 36.04 ATOM 1570 CD2 HIS 362 77.793 11.885 14.224 1.00 34.77 ATOM 1571 ND1 HIS 362 79.50913.212 14.482 1.00 37.16 ATOM 1572 CE1 HIS 362 79.983 11.990 14.325 1.00 37.16 ATOM 1573 NE2 HIS 362 78.962 11.165 14.167 1.00 40.13 ATOM 1574 C HIS 362 75.829 16.110 13.417 1.00 31.00 ATOM 1575 O HIS 362 75.617 16.608 14.525 1.00 30.22 ATOM1576 N PHE 363 75.478 16.690 12.272 1.00 33.06 ATOM 1577 CA PHE 363 74.878 18.021 12.200 1.00 28.08 ATOM 1578 CB PHE 363 74.503 18.355 10.747 1.00 25.26 ATOM 1579 CG PHE 363 73.923 19.733 10.567 1.00 24.91 ATOM 1580 CD1 PHE 363 74.750 20.81710.320 1.00 27.60 ATOM 1581 CD2 PHE 363 72.552 19.948 10.664 1.00 25.52 ATOM 1582 CE1 PHE 363 74.221 22.1.00 10.175 1.00 29.70 ATOM 1583 CE2 PHE 363 72.014 21.227 10.522 1.00 25.88 ATOM 1584 CZ PHE 363 72.850 22.304 10.278 1.00 21.49 ATOM 1585C PHE 363 73.659 18.201 13.099 1.00 23.79 ATOM 1586 O PHE 363 73.587 19.164 13.863 1.00 24.48 ATOM 1587 N TRP 364 72.707 17.277 13.012 1.00 23.13 ATOM 1588 CA TRP 364 71.484 17.369 13.805 1.00 25.06 ATOM 1589 CB TRP 364 70.536 16.201 13.494 1.00 21.17 ATOM 1590 CG TRP 364 69.247 16.220 14.271 1.00 23.14 ATOM 1591 CD2 TRP 364 68.261 17.266 14.296 1.00 27.68 ATOM 1592 CE2 TRP 364 67.229 16.845 15.165 1.00 28.31 ATOM 1593 CE3 TRP 364 68.149 18.517 13.671 1.00 26.46 ATOM 1594 CD1 TRP 36468.784 15.241 15.096 1.00 23.76 ATOM 1595 NE1 TRP 364 67.576 15.607 15.637 1.00 32.12 ATOM 1596 CZ2 TRP 364 66.100 17.628 15.427 1.00 25.63 ATOM 1597 CZ3 TRP 364 67.028 19.294 13.931 1.00 25.55 ATOM 1598 CH2 TRP 364 66.017 18.845 14.803 1.00 29.79 ATOM 1599 C TRP 364 71.715 17.531 15.312 1.00 27.80 ATOM 1600 O TRP 364 71.212 18.486 15.904 1.00 26.96 ATOM 1601 N PRO 365 72.458 16.605 15.955 1.00 30.69 ATOM 1602 CD PRO 365 72.974 15.308 15.481 1.00 31.45 ATOM 1603 CA PRO 365 72.68716.757 17.397 1.00 27.97 ATOM 1604 CB PRO 365 73.506 15.512 17.752 1.00 26.50 ATOM 1605 CG PRO 365 73.057 14.509 16.757 1.00 33.47 ATOM 1606 C PRO 365 73.457 18.043 17.709 1.00 27.10 ATOM 1607 O PRO 365 73.154 18.736 18.681 1.00 26.88 ATOM 1608N LYS 366 74.440 18.365 16.873 1.00 26.99 ATOM 1609 CA LYS 366 75.230 19.577 17.061 1.00 30.69 ATOM 1610 CB LYS 366 76.275 19.708 15.957 1.00 28.53 ATOM 1611 CG LYS 366 77.481 18.804 16.106 1.00 28.89 ATOM 1612 CD LYS 366 78.430 19.027 14.9391.00 32.51

ATOM 1613 CE LYS 366 79.743 18.294 15.116 1.00 38.52 ATOM 1614 NZ LYS 366 80.632 18.506 13.939 1.00 45.28 ATOM 1615 C LYS 366 74.349 20.831 17.079 1.00 36.18 ATOM 1616 O LYS 366 74.472 21.672 17.972 1.00 39.82 ATOM 1617 N LEU 367 73.46420.950 16.091 1.00 37.54 ATOM 1618 CA LEU 367 72.557 22.092 15.994 1.00 36.14 ATOM 1619 CB LEU 367 71.803 22.070 14.659 1.00 32.20 ATOM 1620 CG LEU 367 70.764 23.179 14.447 1.00 36.16 ATOM 1621 CD1 LEU 367 71.402 24.567 14.618 1.00 20.60 ATOM1622 CD2 LEU 367 70.139 23.030 13.065 1.00 34.30 ATOM 1623 C LEU 367 71.561 22.060 17.143 1.00 36.84 ATOM 1624 O LEU 367 71.231 23.091 17.729 1.00 36.94 ATOM 1625 N LEU 368 71.083 20.866 17.459 1.00 37.81 ATOM 1626 CA LEU 368 70.130 20.683 18.5361.00 34.83 ATOM 1627 CB LEU 368 69.763 19.205 18.622 1.00 36.98 ATOM 1628 CG LEU 368 68.421 18.777 19.205 1.00 40.34 ATOM 1629 CD1 LEU 368 67.276 19.595 18.619 1.00 36.28 ATOM 1630 CD2 LEU 368 68.241 17.299 18.908 1.00 39.39 ATOM 1631 C LEU 36870.755 21.182 19.843 1.00 38.32 ATOM 1632 O LEU 368 70.059 21.711 20.707 1.00 41.87 ATOM 1633 N MET 369 72.075 21.057 19.962 1.00 39.46 ATOM 1634 CA MET 369 72.790 21.515 21.154 1.00 40.12 ATOM 1635 CB MET 369 74.219 20.971 21.168 1.00 41.26 ATOM 1636 CG MET 369 74.307 19.493 21.521 1.00 47.83 ATOM 1637 SD MET 369 75.961 18.810 21.289 1.00 55.72 ATOM 1638 CE MET 369 76.809 19.474 22.727 1.00 54.37 ATOM 1639 C MET 369 72.805 23.039 21.251 1.00 42.81 ATOM 1640 O MET 369 72.990 23.60122.335 1.00 47.81 ATOM 1641 N LYS 370 72.622 23.708 20.115 1.00 40.09 ATOM 1642 CA LYS 370 72.588 25.165 20.080 1.00 33.65 ATOM 1643 CB LYS 370 72.751 25.677 18.650 1.00 30.83 ATOM 1644 CG LYS 370 74.138 25.435 18.078 1.00 30.98 ATOM 1645 CDLYS 370 75.188 26.198 18.867 1.00 37.82 ATOM 1646 CE LYS 370 76.591 2S.938 18.351 1.00 36.05 ATOM 1647 NZ LYS 370 77.034 24.562 18.667 1.00 48.68 ATOM 1648 C LYS 370 71.293 25.684 20.702 1.00 33.32 ATOM 1649 O LYS 370 71.218 26.842 21.112 1.00 34.75 ATOM 1650 N VAL 371 70.277 24.826 20.779 1.00 31.90 ATOM 1651 CA VAL 371 69.006 25.197 21.395 1.00 31.77 ATOM 1652 CB VAL 371 67.933 24.092 21.214 1.00 30.28 ATOM 1653 CG1 VAL 371 66.673 24.429 21.995 1.00 30.02 ATOM 1654 CG2 VAL 37167.596 23.933 19.746 1.00 32.23 ATOM 1655 C VAL 371 69.277 25.417 22.885 1.00 34.44 ATOM 1656 O VAL 37L 68.722 26.331 23.499 1.00 33.35 ATOM 1657 N THR 372 70.161 24.590 23.443 1.00 33.1S ATOM 1658 CA THR 372 70.551 24.675 24.847 1.00 32.47 ATOM 1659 CB THR 372 71.541 23.556 25.207 1.00 32.11 ATOM 1660 OG1 THR 372 70.955 22.288 24.891 1.00 35.33 ATOM 1661 CG2 THR 372 71.894 23.603 26.688 1.00 32.54 ATOM 1662 C THR 372 71.226 26.020 25.108 1.00 34.49 ATOM 1663 O THR 372 70.936 26.69626.099 1.00 34.07 ATOM 1664 N ASP 373 72.120 26.405 24.202 1.00 32.77 ATOM 1665 CA ASP 373 72.830 27.671 24.315 1.00 28.08 ATOM 1666 CB ASP 373 73.803 27.841 23.147 1.00 31.59 ATOM 1667 CG ASP 373 74.910 26.789 23.142 1.00 ,37.29 ATOM 1668 OD1ASP 373 75.170 26.169 24.196 1.00 40.82 ATOM 1669 OD2 ASP 373 75.531 26.586 22.079 1.00 40.81 ATOM 1670 C ASP 373 71.830 28.821 24.353 1.00 29.21 ATOM 1671 O ASP 373 71.931 29.709 25.200 1.00 31.85 ATOM 1672 N LEU 374 70.843 28.775 23.463 1.00 24.71 ATOM 1673 CA LEU 374 69.813 29.802 23.403 1.00 25.25 ATOM 1674 CB LEU 374 68.906 29.587 22.188 1.00 25.61 ATOM 1675 CG LEU 374 69.480 30.084 20.858 1.00 25.51 ATOM 1676 CD1 LEU 374 68.741 29.469 19.677 1.00 23.53 ATOM 1677 CD2 LEU 37469.405 31.596 20.820 1.00 21.92 ATOM 1678 C LEU 374 68.994 29.827 24.686 1.00 26.84 ATOM 1679 O LEU 374 68.591 30.895 25.151 1.00 28.96 ATOM 1680 N ARG 375 68.746 28.651 25.254 1.00 31.00 ATOM 1681 CA ARG 375 67.996 28.554 26.502 1.00 32.86 ATOM 1682 CB ARG 375 67.831 27.090 26.924 1.00 36.80 ATOM 1683 CG ARG 375 66.861 26.297 26.071 1.00 44.91 ATOM 1684 CD ARG 375 65.433 26.731 26.338 1.00 58.99 ATOM 1685 NE ARG 375 64.501 26.210 25.342 1.00 72.26 ATOM 1686 CZ ARG 375 63.909 25.02025.404 1.00 77.46 ATOM 1687 NH1 ARG 375 64.147 24.201 26.422 1.00 80.94 ATOM 1688 NH2 ARG 375 63.062 24.657 24.447 1.00 75.58 ATOM 1689 C ARG 375 68.771 29.317 27.570 1.00 32.27 ATOM 1690 O ARG 375 68.199 30.125 28.304 1.00 33.75 ATOM 1691 NMET 376 70.084 29.098 27.602 1.00 32.65 ATOM 1692 CA MET 376 70.967 29.753 28.560 1.00 35.83 ATOM 1693 CB MET 376 72.392 29.210 28.434 1.00 39.25 ATOM 1694 CG MET 376 72.526 27.751 28.839 1.00 54.45 ATOM 1695 SD MET 376 74.245 27.212 28.944 1.00 73.93 ATOM 1696 CE MET 376 74.421 26.270 27.434 1.00 67.01 ATOM 1697 C MET 376 70.960 31.267 28.378 1.00 35.38 ATOM 1698 O MET 376 70.882 32.015 29.353 1.00 34.73 ATOM 1699 N ILE 377 71.038 31.716 27.129 1.00 32.51 ATOM 1700 CA ILE 377 71.01633.142 26.816 1.00 26.55 ATOM 1701 CB ILE 377 71.182 33.370 25.299 1.00 24.84 ATOM 1702 CG2 ILE 377 70.817 34.797 24.923 1.00 26.63 ATOM 1703 CG1 ILE 377 72.616 33.038 24.890 1.00 20.66 ATOM 1704 CD1 ILE 377 72.872 33.104 23.409 1.00 20.74 ATOM1705 C ILE 377 69.706 33.755 27.313 1.00 25.47 ATOM 1706 O ILE 377 69.696 34.848 27.881 1.00 29.99 ATOM 1707 N GLY 378 68.608 33.033 27.127 1.00 25.11 ATOM 1708 CA GLY 378 67.321 33.522 27.580 1.00 27.82 ATOM 1709 C GLY 378 67.279 33.613 29.0951.00 30.90 ATOM 1710 O GLY 378 66.740 34.5.79 29.651 1.00 31.19 ATOM 1711 N ALA 379 67.851 32.611 29.761 1.00 31.62 ATOM 1712 CA ALA 379 67.896 32.547 31.223 1.00 30.74 ATOM 1713 CB ALA 379 68.433 31.198 31.671 1.00 30.82 ATOM 1714 C ALA 37968.756 33.668 31.801 1.00 30.07 ATOM 1715 O ALA 379 68.327 34.384 32.708 1.00 31.05 ATOM 1716 N CYA 380 69.966 33.817 31.273 1.00 29.72 ATOM 1717 CA CYA 380 70.873 34.866 31.723 1.00 33.36 ATOM 1718 CB CYA 380 72.201 34.809 30.963 1.00 38.31 ATOM 1719 SG CYA 380 73.249 33.407 31.386 1.00 50.99 ATOM 1720 AS CYA 380 74.982 33.655 29.929 1.00 70.37 ATOM 1721 C CYA 380 70.226 36.232 31.535 1.00 33.40 ATOM 1722 O CYA 380 70.246 37.062 32.442 1.00 36.41 ATOM 1723 N HIS 381 69.615 36.45630.374 1.00 32.55 ATOM 1724 CA HIS 381 68.965 37.734 30.114 1.00 26.41 ATOM 1725 CB HIS 381 68.434 37.811 28.681 1.00 20.89 ATOM 1726 CG HIS 381 67.593 39.023 28.423 1.00 15.78 ATOM 1727 CD2 HIS 381 67.928 40.277 28.041 1.00 12.67 ATOM 1728 ND1HIS 381 66.226 39.031 28.605 1.00 17.88 ATOM 1729 CE1 HIS 381 65.756 40.239 28.353 1.00 16.27 ATOM 1730 NE2 HIS 381 66.768 41.013 28.008 1.00 17.18 ATOM 1731 C HIS 381 67.839 38.023 31.102 1.00 26.73 ATOM 1732 O HIS 381 67.621 39.176 31.464 1.00 30.46 ATOM 1733 N ALA 382 67.111 36.991 31.521 1.00 26.68 ATOM 1734 CA ALA 382 66.010 37.176 32.464 1.00 27.90 ATOM 1735 CB ALA 382 65.237 35.878 32.642 1.00 25.29 ATOM 1736 C ALA 382 66.511 37.697 33.810 1.00 31.23 ATOM 1737 O ALA 382 65.92738.617 34.378 1.00 37.67 ATOM 1738 N SER 383 67.596 37.114 34.316 1.00

34.15 ATOM 1739 CA SER 383 68.174 37.550 35.588 1.00 37.23 ATOM 1740 CB SER 383 69.294 36.605 36.027 1.00 40.21 ATOM 1741 OG SER 383 68.785 35.324 36.361 1.00 53.99 ATOM 1742 C SER 383 68.727 38.958 35.417 1.00 33.67 ATOM 1743 O SER 38368.532 39.827 36.268 1.00 40.73 ATOM 1744 N ARG 384 69.411 39.171 34.298 1.00 29.95 ATOM 1745 CA ARG 384 70.000 40.458 33.957 1.00 29.77 ATOM 1746 CB ARG 384 70.684 40.350 32.594 1.00 30.79 ATOM 1747 CG ARG 384 71.481 41.558 32.167 1.00 31.34 ATOM 1748 CD ARG 384 72.781 41.638 32.918 1.00 33.62 ATOM 1749 NE ARG 384 73.657 42.660 32.358 1.00 41.68 ATOM 1750 CZ ARG 384 74.584 43.310 33.052 1.00 41.20 ATOM 1751 NH1 ARG 384 74.756 43.047 34.339 1.00 42.11 ATOM 1752 NH2 ARG 384 75.34944.213 32.455 1.00 37.27 ATOM 1753 C ARG 384 68.910 41.536 33.911 1.00 35.72 ATOM 1754 O ARG 384 69.090 42.635 34.439 1.00 41.66 ATOM 1755 N PHE 385 67.768 41.196 33.318 1.00 34.30 ATOM 1756 CA PHE 385 66.646 42.119 33.199 1.00 32.40 ATOM 1757CB PHE 385 65.527 41.502 32.356 1.00 29.02 ATOM 1758 CG PHE 385 64.344 42.407 32.163 1.00 26.56 ATOM 1759 CD1 PHE 385 64.317 43.320 31.119 1.00 26.59 ATOM 1760 CD2 PHE 385 63.263 42.355 33.037 1.00 24.69 ATOM 1761 CE1 PHE 385 63.231 44.173 30.9471.00 31.70 ATOM 1762 CE2 PHE 385 62.174 43.202 32.875 1.00 26.79 ATOM 1763 CZ PHE 385 62.158 44.115 31.827 1.00 31.59 ATOM 1764 C PHE 385 66.121 42.492 34.578 1.00 32.98 ATOM 1765 O PHE 385 65.822 43.659 34.839 1.00 33.91 ATOM 1766 N LEU 38666.003 41.499 35.456 1.00 33.91 ATOM 1767 CA LEU 386 65.533 41.736 36.818 1.00 38.66 ATOM 1768 CB LEU 386 65.547 40.440 37.633 1.00 43.79 ATOM 1769 CG LEU 386 64.327 39.521 37.525 1.00 49.81 ATOM 1770 CD1 LEU 386 64.652 38.147 38.099 1.00 51.12 ATOM 1771 CD2 LEU 386 63.135 40.148 38.246 1.00 49.17 ATOM 1772 C LEU 386 66.445 42.761 37.475 1.00 38.95 ATOM 1773 O LEU 386 65.979 43.682 38.146 1.00 42.16 ATOM 1774 N HIS 387 67.745 42.613 37.248 1.00 33.62 ATOM 1775 CA HIS 387 68.723 43.53137.808 1.00 39.73 ATOM 1776 CB HIS 387 70.138 42.980 37.639 1.00 40.71 ATOM 1777 CG HIS 387 70.403 41.749 38.449 1.00 52.03 ATOM 1778 CD2 HIS 387 69.573 40.967 39.181 1.00 53.85 ATOM 1779 ND1 HIS 387 71.657 41.189 38.566 1.00 54.79 ATOM 1780CE1 HIS 387 71.590 40.114 39.334 1.00 56.55 ATOM 1781 NE2 HIS 387 70.336 39.958 39.720 1.00 57.48 ATOM 1782 C HIS 387 68.594 44.913 37.175 1.00 42.08 ATOM 1783 O HIS 387 68.712 45.926 37.865 1.00 44.12 ATOM 1784 N MET 388 68.318 44.957 35.8741.00 42.38 ATOM 1785 CA MET 388 68.154 46.229 35.175 1.00 38.00 ATOM 1786 CB MET 388 67.840 46.006 33.692 1.00 40.21 ATOM 1787 CG MET 388 69.009 45.555 32.829 1.00 41.26 ATOM 1788 SD MET 388 68.500 45.427 31.089 1.00 45.51 ATOM 1789 CE MET 38869.089 43.802 30.645 1.00 42.40 ATOM 1790 C MET 388 67.025 47.044 35.810 1.00 38.11 ATOM 1791 O MET 388 67.155 48.255 35.997 1.00 38.41 ATOM 1792 N LYS 389 65.926 46.374 36.144 1.00 39.67 ATOM 1793 CA LYS 389 64.773 47.036 36.750 1.00 44.96 ATOM 1794 CB LYS 389 63.570 46.087 36.818 1.00 49.52 ATOM 1795 CG LYS 389 62.674 46.102 35.588 1.00 56.74 ATOM 1796 CD LYS 389 62.145 47.509 35.278 1.00 68.05 ATOM 1797 CE LYS 389 61.287 48.100 36.403 1.00 71.47 ATOM 1798 NZ LYS 389 60.038 47.33036.661 1.00 71.98 ATOM 1799 C LYS 389 65.041 47.604 38.141 1.00 46.60 ATOM 1800 O LYS 389 64.516 48.661 38.499 1.00 47.25 ATOM 1801 N VAL 390 65.832 46.893 38.935 1.00 47.15 ATOM 1802 CA VAL 390 66.129 47.353 40.284 1.00 50.75 ATOM 1803 CB VAL390 66.686 46.202 41.182 1.00 50.42 ATOM 1804 CG1 VAL 390 68.095 45.802 40.770 1.00 47.93 ATOM 1805 CG2 VAL 390 66.650 46.612 42.640 1.00 56.67 ATOM 1806 C VAL 390 67.072 48.558 40.286 1.00 49.82 ATOM 1807 O VAL 390 66.971 49.426 41.152 1.00 52.44 ATOM 1808 N GLU 391 67.926 48.651 39.272 1.00 46.14 ATOM 1809 CA GLU 391 68.888 49.741 39.173 1.00 43.84 ATOM 1810 CB GLU 391 70.150 49.268 38.449 1.00 41.44 ATOM 1811 CG GLU 391 70.837 48.074 39.095 1.00 51.12 ATOM 1812 CD GLU 391 71.21848.325 40.540 1.00 57.29 ATOM 1813 OE1 GLU 391 71.970 49.287 40.802 1.00 58.15 ATOM 1814 OE2 GLU 391 70.764 47.559 41.416 1.00 62.51 ATOM 1815 C GLU 391 68.386 51.015 38.501 1.00 45.94 ATOM 1816 O GLU 391 68.567 52.114 39.033 1.00 51.14 ATOM1817 N CYA 392 67.727 50.872 37.354 1.00 45.84 ATOM 1818 CA CYA 392 67.255 52.029 36.598 1.00 41.60 ATOM 1819 CB CYA 392 67.681 51.889 35.140 1.00 42.06 ATOM 1820 SG CYA 392 69.452 52.008 34.968 1.00 44.47 ATOM 1821 AS CYA 392 69.867 50.81233.150 1.00 54.22 ATOM 1822 C CYA 392 65.779 52.395 36.683 1.00 42.27 ATOM 1823 O CYA 392 64.937 51.564 37.029 1.00 43.91 ATOM 1824 N PRO 393 65.451 53.674 36.414 1.00 42.79 ATOM 1825 CD PRO 393 66.384 54.774 36.106 1.00 38.59 ATOM 1826 CA PRO393 64.067 54.159 36.459 1.00 44.20 ATOM 1827 CB PRO 393 64.218 55.667 36.238 1.00 39.88 ATOM 1828 CG PRO 393 65.487 55.789 35.459 1.00 35.88 ATOM 1829 C PRO 393 63.178 53.513 35.398 1.00 45.29 ATOM 1830 O PRO 393 63.600 53.308 34.257 1.00 43.97 ATOM 1831 N THR 394 61.935 53.238 35.782 1.00 48.20 ATOM 1832 CA THR 394 60.959 52.607 34.901 1.00 53.71 ATOM 1833 CB THR 394 59.605 52.429 35.629 1.00 59.59 ATOM 1834 OG1 THR 394 58.690 51.717 34.787 1.00 66.50 ATOM 1835 CG2 THR 394 59.01353.787 36.004 1.00 61.00 ATOM 1836 C THR 394 60.752 53.358 33.581 1.00 51.35 ATOM 1837 O THR 394 60.419 52.751 32.563 1.00 54.39 ATOM 1838 N GLU 395 61.008 54.664 33.595 1.00 47.65 ATOM 1839 CA GLU 395 60.845 55.509 32.414 1.00 44.43 ATOM 1840CB GLU 395 60.988 56.978 32.804 1.00 43.85 ATOM 1841 C GLU 395 61.788 55.175 31.250 1.00 42.93 ATOM 1842 O GLU 395 61.589 55.649 30.129 1.00 41.39 ATOM 1843 N LEU 396 62.818 54.375 31.517 1.00 39.38 ATOM 1844 CA LEU 396 63.782 53.989 30.486 1.00 35.70 ATOM 1845 CB LEU 396 65.185 53.867 31.090 1.00 34.96 ATOM 1846 CG LEU 396 65.854 55.141 31.609 1.00 36.47 ATOM 1847 CD1 LEU 396 67.234 54.807 32.150 1.00 34.21 ATOM 1848 CD2 LEU 396 65.959 56.164 30.491 1.00 32.74 ATOM 1849 C LEU 39663.407 52.671 29.803 1.00 34.60 ATOM 1850 O LEU 396 64.086 52.223 28.873 1.00 30.36 ATOM 1851 N PHE 397 62.325 52.059 30.269 1.00 33.02 ATOM 1852 CA PHE 397 61.868 50.792 29.725 1.00 33.39 ATOM 1853 CB PHE 397 61.615 49.782 30.852 1.00 34.30 ATOM 1854 CG PHE 397 62.834 49.439 31.665 1.00 32.62 ATOM 1855 CD1 PHE 397 63.296 50.301 32.654 1.00 32.35 ATOM 1856 CD2 PHE 397 63.504 48.241 31.461 1.00 31.28 ATOM 1857 CE1 PHE 397 64.407 49.976 33.426 1.00 27.01 ATOM 1858 CE2 PHE 397 64.61647.905 32.229 1.00 33.34 ATOM 1859 CZ PHE 397 65.067 48.775 33.213 1.00 31.29 ATOM 1860 C PHE 397 60.580 50.961 28.934 1.00 33.17 ATOM 1861 O PHE 397 59.540 51.318 29.498 1.00 31.99 ATOM 1862 N PRO 398 60.636 50.752 27.606 1.00 32.45 ATOM 1863CD PRO 398 61.821 50.493 26.768 1.00 28.15

ATOM 1864 CA PRO 398 59.429 50.885 26.786 1.00 30.02 ATOM 1865 CB PRO 398 59.921 50.483 25.394 1.00 28.15 ATOM 1866 CG PRO 398 61.352 50.923 25.397 1.00 24.89 ATOM 1867 C PRO 398 58.384 49.900 27.326 1.00 28.39 ATOM 1868 O PRO 398 58.73548.810 27.789 1.00 28.00 ATOM 1869 N PRO 399 57.092 50.262 27.267 1.00 32.45 ATOM 1870 CD PRO 399 56.577 51.511 26.672 1.00 34.93 ATOM 1871 CA PRO 399 55.989 49.421 27.753 1.00 32.54 ATOM 1872 CB PRO 399 54.755 50.122 27.188 1.00 34.47 ATOM1873 CG PRO 399 55.159 51.564 27.196 1.00 31.37 ATOM 1874 C PRO 399 56.044 47.946 27.338 1.00 32.18 ATOM 1875 O PRO 399 55.950 47.054 28.188 1.00 32.58 ATOM 1876 N LEU 400 56.195 47.689 26.041 1.00 30.15 ATOM 1877 CA LEU 400 56.259 46.314 25.5411.00 32.32 ATOM 1878 CB LEU 400 56.211 46.297 24.011 1.00 28.67 ATOM 1879 CG LEU 400 56.028 44.927 23.351 1.00 28.77 ATOM 1880 CD1 LEU 400 54.802 44.234 23.919 1.00 22.73 ATOM 1881 CD2 LEU 400 55.897 45.096 21.846 1.00 27.89 ATOM 1882 C LEU 40057.496 45.561 26.051 1.00 32.27 ATOM 1883 O LEU 400 57.437 44.358 26.307 1.00 32.87 ATOM 1884 N PHE 401 58.602 46.279 26.215 1.00 32.27 ATOM 1885 CA PHE 401 59.847 45.695 26.710 1.00 32.39 ATOM 1886 CB PHE 401 60.946 46.769 26.711 1.00 31.38 ATOM 1887 CG PHE 401 62.290 46.286 27.194 1.00 35.12 ATOM 1888 CD1 PHE 401 62.835 45.089 26.729 1.00 34.68 ATOM 1889 CD2 PHE 401 63.030 47.051 28.097 1.00 34.57 ATOM 1890 CE1 PHE 401 64.100 44.662 27.155 1.00 30.27 ATOM 1891 CE2 PHE 401 64.29146.635 28.526 1.00 33.57 ATOM 1892 CZ PHE 401 64.828 45.438 28.054 1.00 35.74 ATOM 1893 C PHE 401 59.599 45.169 28.129 1.00 32.21 ATOM 1894 O PHE 401 60.002 44.056 28.478 1.00 33.36 ATOM 1895 N LEU 402 58.902 45.967 28.929 1.00 31.85 ATOM 1896CA LEU 402 58.582 45.602 30.302 1.00 35.06 ATOM 1897 CB LEU 402 57.948 46.789 31.029 1.00 34.76 ATOM 1898 CG LEU 402 58.878 47.852 31.591 1.00 33.48 ATOM 1899 CD1 LEU 402 58.060 49.010 32.152 1.00 32.58 ATOM 1900 CD2 LEU 402 59.753 47.217 32.6621.00 26.27 ATOM 1901 C LEU 402 57.626 44.426 30.393 1.00 36.80 ATOM 1902 O LEU 402 57.793 43.545 31.239 1.00 35.43 ATOM 1903 N GLU 403 56.600 44.443 29.547 1.00 38.50 ATOM 1904 CA GLU 403 55.581 43.401 29.540 1.00 40.24 ATOM 1905 CB GLU 40354.435 43.792 28.605 1.00 44.03 ATOM 1906 CG GLU 403 53.239 42.850 28.666 1.00 55.53 ATOM 1907 CD GLU 403 52.180 43.159 27.618 1.00 66.67 ATOM 1908 OE1 GLU 403 52.151 44.299 27.095 1.00 70.81 ATOM 1909 OE2 GLU 403 51.370 42.255 27.315 1.00 73.80 ATOM 1910 C GLU 403 56.096 42.018 29.162 1.00 38.00 ATOM 1911 O GLU 403 55.745 41.029 29.805 1.00 38.78 ATOM 1912 N VAL 404 56.934 41.955 28.132 1.00 37.39 ATOM 1913 CA VAL 404 57.475 40.686 27.652 1.00 37.05 ATOM 1914 CB VAL 404 58.180 40.85526.286 1.00 35.57 ATOM 1915 CG1 VAL 404 58.677 39.513 25.776 1.00 36.85 ATOM 1916 CG2 VAL 404 57.222 41.451 25.287 1.00 42.03 ATOM 1917 C VAL 404 58.438 40.000 28.609 1.00 38.69 ATOM 1918 O VAL 404 58.436 38.774 28.727 1.00 40.71 ATOM 1919 NPHE 405 59.267 40.785 29.286 1.00 39.34 ATOM 1920 CA PHE 405 60.250 40.221 30.198 1.00 39.33 ATOM 1921 CB PHE 405 61.620 40.840 29.913 1.00 33.87 ATOM 1922 CG PHE 405 62.107 40.609 28.509 1.00 32.17 ATOM 1923 CD1 PHE 405 62.355 41.683 27.660 1.00 31.34 ATOM 1924 CD2 PHE 405 62.315 39.317 28.032 1.00 31.98 ATOM 1925 CE1 PHE 405 62.801 41.476 26.352 1.00 30.79 ATOM 1926 CE2 PHE 405 62.759 39.099 26.730 1.00 26.06 ATOM 1927 CZ PHE 405 63.004 40.182 25.889 1.00 27.98 ATOM 1928 C PHE 40559.905 40.322 31.682 1.00 42.64 ATOM 1929 O PHE 405 60.785 40.188 32.534 1.00 45.10 ATOM 1930 N GLU 406 58.630 40.536 31.988 1.00 48.95 ATOM 1931 CA GLU 406 58.181 40.641 33.373 1.00 56.93 ATOM 1932 CB GLU 406 56.820 41.324 33.432 1.00 56.94 ATOM 1933 C GLU 406 58.116 39.263 34.040 1.00 61.92 ATOM 1934 O GLU 406 57.988 38.256 33.308 1.00 67.61 ATOM 1 O1 HOH 501 67.588 36.828 11.225 1.00 27.32 ATOM 2 O1 HOH 502 68.647 41.203 12.940 1.00 39.54 ATOM 3 O1 HOH 503 64.072 40.115 12.4071.00 32.47 ATOM 4 O1 HOH 504 62.312 39.659 16.075 1.00 17.39 ATOM 5 O1 HOH 505 63.449 46.468 15.530 1.00 30.46 ATOM 6 O1 HOH 506 67.191 15.561 -0.279 1.00 35.96 ATOM 7 O1 HOH 507 67.100 11.855 0.295 1.00 20.00 ATOM 8 O1 HOH 508 61.004 15.5100.047 1.00 20.00 ATOM 9 O1 HOH 509 59.851 10.761 6.050 1.00 20.00 ATOM 10 O1 HOH 510 57.553 11.824 10.360 1.00 44.63 ATOM 11 O1 HOH 511 54.101 13.545 8.720 1.00 20.00 ATOM 12 O1 HOH 512 55.923 15.916 12.205 1.00 29.31 ATOM 13 O1 HOH 513 50.90019.934 8.193 1.00 20.00 ATOM 14 O1 HOH 514 50.474 22.912 7.942 1.00 45.34 ATOM 15 O1 HOH 515 49.737 20.631 11.530 1.00 20.00 ATOM 16 O1 HOH 516 50.829 25.467 13.330 1.00 20.00 ATOM 17 O1 HOH 517 53.818 25.833 10.682 1.00 42.12 ATOM 18 O1 HOH518 52.591 31.216 7.313 1.00 35.55 ATOM 19 O1 HOH 519 58.510 31.667 2.158 1.00 20.00 ATOM 20 O1 HOH 520 58.235 36.751 2.232 1.00 20.00 ATOM 21 O1 HOH 521 62.484 37.992 5.537 1.00 20.00 ATOM 22 O1 HOH 522 68.184 36.969 5.889 1.00 50.08 ATOM 23O1 HOH 523 66.889 33.781 8.584 1.00 20.00 ATOM 24 O1 HOH 524 67.217 30.836 3.085 1.00 34.44 ATOM 25 O1 HOH 525 64.336 28.325 3.098 1.00 20.00 ATOM 26 O1 HOH 526 67.667 26.625 1.519 1.00 20.00 ATOM 27 O1 HOH 527 76.757 22.883 5.467 1.00 36.94 ATOM 28 O1 HOH 528 72.250 17.936 6.950 1.00 36.00 ATOM 29 O1 HOH 529 71.760 14.791 8.058 1.00 40.18 ATOM 30 O1 HOH 530 72.884 14.751 11.484 1.00 41.44 ATOM 31 O1 HOH 531 69.235 12.986 11.709 1.00 39.38 ATOM 32 O1 HOH 532 69.402 12.036 14.891 1.00 40.68 ATOM 33 O1 HOH 533 64.560 10.910 15.076 1.00 20.00 ATOM 34 O1 HOH 534 63.169 10.413 11.722 1.00 20.00 ATOM 35 O1 HOH 535 66.042 11.455 11.077 1.00 41.05 ATOM 36 O1 HOH 536 76.285 12.458 10.677 1.00 20.00 ATOM 37 O1 HOH 537 81.094 22.52013.435 1.00 48.70 ATOM 38 O1 HOH 538 80.505 25.457 14.849 1.00 46.30 ATOM 39 O1 HOH 539 77.669 21.932 18.119 1.00 43.79 ATOM 40 O1 HOH 540 77.187 28.903 21.137 1.00 40.22 ATOM 41 O1 HOH 541 76.420 30.760 23.658 1.00 29.63 ATOM 42 O1 HOH 54283.028 32.743 20.922 1.00 38.14 ATOM 43 O1 HOH 543 82.842 43.133 17.983 1.00 39.36 ATOM 44 O1 HOH 544 77.484 34.040 9.664 1.00 36.37 ATOM 45 O1 HOH 545 75.904 32.986 12.256 1.00 34.93 ATOM 46 O1 HOH 546 74.185 29.689 9.761 1.00 38.60 ATOM 47 O1HOH 547 64.936 20.644 23.365 1.00 36.83 ATOM 48 O1 HOH 548 61.750 22.313 25.288 1.00 34.81 ATOM 49 O1 HOH 549 59.544 21.463 26.162 1.00 20.00 ATOM 50 O1 HOH 550 62.300 27.528 24.386 1.00 35.89 ATOM 51 O1 HOH 551 58.228 29.424 24.603 1.00 25.47 ATOM 52 O1 HOH 552 57.368 32.196 30.527 1.00 45.27 ATOM 53 O1 HOH 553 62.063 36.304 30.245 1.00 42.26 ATOM 54 O1 HOH 554 64.722 36.725 28.906 1.00 24.66 ATOM 55 O1 HOH 555 62.207 35.851 26.642 1.00

30.36 ATOM 56 O1 HOH 556 63.608 33.715 25.707 1.00 42.74 ATOM 57 O1 HOH 557 62.979 38.422 32.977 1.00 49.93 ATOM 58 O1 HOH 558 66.911 33.364 34.901 1.00 50.02 ATOM 59 O1 HOH 559 72.608 29.636 31.674 1.00 37.60 ATOM 60 O1 HOH 560 76.96740.633 32.514 1.00 44.81 ATOM 61 O1 HOH 561 73.613 41.817 36.847 1.00 31.79 ATOM 62 O1 HOH 562 75.773 46.227 30.514 1.00 29.06 ATOM 63 O1 HOH 563 79.903 46.178 30.800 1.00 41.67 ATOM 64 O1 HOH 564 69.746 51.175 33.564 1.00 20.00 ATOM 65 O1 HOH565 74.320 52.047 39.438 1.00 20.00 ATOM 66 O1 HOH 566 65.900 53.647 27.404 1.00 40.45 ATOM 67 O1 HOH 567 68.848 53.076 17.895 1.00 39.25 ATOM 68 O1 HOH 568 63.507 48.672 13.581 1.00 43.77 ATOM 69 O1 HOH 569 64.625 46.825 10.331 1.00 20.00 ATOM70 O1 HOH 570 55.882 41.431 11.148 1.00 20.00 ATOM 71 O1 HOH 571 52.830 43.513 20.032 1.00 35.18 ATOM 72 O1 HOH 572 56.990 49.485 24.052 1.00 37.30 ATOM 73 O1 HOH 573 54.188 47.024 30.900 1.00 52.93 ATOM 74 O1 HOH 574 57.823 44.590 34.025 1.00 53.64 ATOM 75 O1 HOH 575 47.827 29.597 30.690 1.00 37.61 ATOM 76 O1 HOH 576 53.030 24.901 32.732 1.00 45.06 ATOM 77 O1 HOH 577 47.569 19.105 28.647 1.00 38.88 ATOM 78 O1 HOH 578 47.232 20.282 25.561 1.00 20.00 ATOM 79 O1 HOH 579 51.960 14.86925.534 1.00 49.45 ATOM 80 O1 HOH 580 52.831 23.395 1.634 1.00 20.00 ATOM 81 O1 HOH 581 51.472 22.968 -0.900 1.00 25.10 ATOM 82 O1 HOH 582 77.238 52.503 8.906 1.00 47.05 ATOM 2004 C1 DMT 1 67.320 42.326 18.648 1.00 28.58 ATOM 2005 C2 DMT 168.927 43.263 23.318 1.00 29.26 ATOM 2006 C3 DMT 1 67.236 43.583 19.236 1.00 24.54 ATOM 2007 C4 DMT 1 69.268 44.313 24.111 1.00 28.48 ATOM 2008 C5 DMT 1 68.003 43.859 20.363 1.00 28.76 ATOM 2009 C6 DMT 1 68.654 44.389 25.458 1.00 28.16 ATOM2010 C7 DMT 1 68.811 42.902 20.875 1.00 26.80 ATOM 2011 C8 DMT 1 67.803 43.410 25.793 1.00 29.83 ATOM 2012 C9 DMT 1 68.921 41.665 20.324 1.00 26.77 ATOM 2013 C10 DMT 1 67.464 42.358 24.989 1.00 28.60 ATOM 2014 C11 DMT 1 68.165 41.349 19.185 1.00 25.29 ATOM 2015 C12 DMT 1 68.059 42.281 23.675 1.00 26.74 ATOM 2016 C13 DMT 1 66.475 42.038 17.456 1.00 21.51 ATOM 2017 C14 DMT 1 68.916 45.478 26.380 1.00 21.05 ATOM 2018 C15 DMT 1 66.989 40.910 16.417 1.00 22.84 ATOM 2019 C16 DMT 1 68.09046.870 26.009 1.00 19.41 ATOM 2020 C17 DMT 1 65.982 40.730 15.243 1.00 27.07 ATOM 2021 C18 DMT 1 70.279 46.131 26.085 1.00 16.03 ATOM 2022 C19 DMT 1 67.903 45.249 20.974 1.00 19.56 ATOM 2023 C20 DMT 1 69.853 40.599 20.901 1.00 4.52 ATOM 2024 N1DMT 1 68.280 41.070 16.042 1.00 17.57 ATOM 2025 O1 DMT 1 67.209 43.465 27.087 1.00 25.94 ATOM 2026 O2 DMT 1 69.547 43.191 22.015 1.00 30.23 ATOM 2027 O3 DMT 1 66.449 40.778 14.118 1.00 29.45 ATOM 2028 O4 DMT 1 64.820 40.564 15.546 1.00 26.46 END

APPENDIX 4 TR_TRIAC.PDB REMARK REMARK TR_triac full length numbering REMARK Rfactor 0.236 Rfree 0.241 REMARK Resolution 25. 2.5 all reflections REMARK REMARK Three cacodylate-modified cysteines: REMARK Cys334, Cys380, Cys392 REMARKmodeled as free arsenic atoms REMARK REMARK conserved polar HOH numbered as in TR_t3.pdb REMARK rearrangements start 600 REMARK REMARK side chain of certain residues modeled as ALA due to poor density; REMARK however, residue name reflects trueresidue for clarity REMARK REMARK clone obtained from Murray et. al. REMARK deposited sequence confirmed, REMARK differing from that reported by Thompson et. al. REMARK in the following codons: REMARK 281 Thr--Ala REMARK 285 Lys--Glu REMARKidentical to that reported by Mitsuhashi et. al. REMARK gb:RNTRAVI X07409 JRNL AUTH M. B. MURRAY, N. D. ZILZ, N. L. MCCREARY, M. J. MACDONALD JRNL AUTH 2 H. C. TOWLE JRNL TITL ISOLATION AND CHARACTERIZATION OF RAT CDNA CLONES FOR TWO JRNL TITL 2DISTINCT THYROID HORMONE RECPTORS JRNL REF JBC V. 263 25 1988 JRNL AUTH C. C. THOMPSON, C. WEINBERGER, R. LEBO, R. M. EVANS JRNL TITL IDENTIFICATION OF A NOVEL THYROID HORMONE RECEPTOR EXPRESSED JRNL TITL 2 IN THE MAMMALIAN CENTRAL NERVOUS SYSTEM JRNL REF SCIENCE V. 237 1987 JRNL AUTH T. MITSUHASHI, G. TENNYSON, V. NIKODEM JRNL TITL NUCLEOTIDE SEQUENCE OF NOVEL CDNAS GENERATED BY ALTERNATIVE JRNL TITL 2 SPLICING OF A RAT THYROID HORMONE RECEPTOR GENE TRANSCRIPT JRNL REF NUC. ACIDS. RES. V.16 12 1988 REMARK ATOM 1 CB ARG 157 9.880 -24.199 7.196 1.00 57.79 ATOM 2 CG ARG 157 11.380 -24.411 7.340 1.00 57.79 ATOM 3 CD ARG 157 11.960 -23.602 8.486 1.00 57.79 ATOM 4 NE ARG 157 11.492 -24.098 9.778 1.00 57.79 ATOM 5 CZ ARG 157 12.284-24.379 10.809 1.00 57.79 ATOM 6 NH1 ARG 157 13.598 -24.212 10.714 1.00 57.79 ATOM 7 NH2 ARG 157 11.762 -24.854 11.932 1.00 57.79 ATOM 8 C ARG 157 7.774 -24.838 5.974 1.00 38.50 ATOM 9 O ARG 157 7.553 -24.416 4.840 1.00 57.79 ATOM 10 N ARG 1579.929 -25.500 5.089 1.00 38.50 ATOM 11 CA ARG 157 9.183 -25.276 6.360 1.00 38.50 ATOM 12 N PRO 158 6.802 -24.951 6.895 1.00 23.08 ATOM 13 CD PRO 158 6.945 -25.424 8.282 1.00 28.38 ATOM 14 CA PRO 158 5.415 -24.562 6.617 1.00 23.08 ATOM 15 CB PRO158 4.704 -24.824 7.948 1.00 28.38 ATOM 16 CG PRO 158 5.801 -24.735 8.966 1.00 28.38 ATOM 17 C PRO 158 5.210 -23.124 6.132 1.00 23.08 ATOM 18 O PRO 158 5.678 -22.167 6.753 1.00 28.38 ATOM 19 N GLU 159 4.504 -23.000 5.012 1.00 19.26 ATOM 20 CAGLU 159 4.191 -21.717 4.389 1.00 19.26 ATOM 21 CB GLU 159 4.022 -21.912 2.878 1.00 24.58 ATOM 22 CG GLU 159 5.317 -22.009 2.086 1.00 24.58 ATOM 23 CD GLU 159 5.849 -20.651 1.659 1.00 24.58 ATOM 24 OE1 GLU 159 5.034 -19.722 1.476 1.00 24.58 ATOM25 OE2 GLU 159 7.080 -20.513 1.490 1.00 24.58 ATOM 26 C GLU 159 2.879 -21.193 4.968 1.00 19.26 ATOM 27 O GLU 159 2.152 -21.931 5.636 1.00 24.58 ATOM 28 N PRO 160 2.579 -19.899 4.765 1.00 17.44 ATOM 29 CD PRO 160 3.442 -18.817 4.259 1.00 13.94 ATOM 30 CA PRO 160 1.323 -19.360 5.299 1.00 17.44 ATOM 31 CB PRO 160 1.414 -17.872 4.956 1.00 13.94 ATOM 32 CG PRO 160 2.880 -17.604 4.952 1.00 13.94 ATOM 33 C PRO 160 0.098 -20.006 4.639 1.00 17.44 ATOM 34 O PRO 160 0.067 -20.207 3.423 1.00 13.94 ATOM 35 N THR 161 -0.895 -20.352 5.450 1.00 17.00 ATOM 36 CA THR 161 -2.119 -20.957 4.941 1.00 17.00 ATOM 37 CB THR 161 -2.958 -21.587 6.086 1.00 20.43 ATOM 38 OG1 THR 161 -3.441 -20.557 6.959 1.00 20.43 ATOM 39 CG2 THR 161 -2.121 -22.5766.888 1.00 20.43 ATOM 40 C THR 161 -2.929 -19.843 4.284 1.00 17.00 ATOM 41 O THR 161 -2.691 -18.660 4.547 1.00 20.43 ATOM 42 N PRO 162 -3.918 -20.200 3.449 1.00 12.94 ATOM 43 CD PRO 162 -4.311 -21.559 3.038 1.00 17.56 ATOM 44 CA PRO 162 -4.743-19.190 2.780 1.00 12.94 ATOM 45 CB PRO 162 -5.846 -20.029 2.143 1.00 17.56 ATOM 46 CG PRO 162 -5.147 -21.303 1.816 1.00 17.56 ATOM 47 C PRO 162 -5.317 -18.171 3.763 1.00 12.94 ATOM 48 O PRO 162 -5.305 -16.964 3.503 1.00 17.56 ATOM 49 N GLU 163-5.790 -18.668 4.903 1.00 19.45 ATOM 50 CA GLU 163 -6.374 -17.828 5.943 1.00 19.45 ATOM 51 CB GLU 163 -6.994 -18.690 7.047 1.00 49.96 ATOM 52 CG GLU 163 -8.178 -19.558 6.606 1.00 49.96 ATOM 53 CD GLU 163 -7.782 -20.720 5.697 1.00 49.96 ATOM 54OE1 GLU 163 -6.735 -21.361 5.951 1.00 49.96 ATOM 55 OE2 GLU 163 -8.527 -20.999 4.731 1.00 49.96 ATOM 56 C GLU 163 -5.330 -16.897 6.548 1.00 19.45 ATOM 57 O GLU 163 -5.614 -15.731 6.832 1.00 49.96 ATOM 58 N GLU 164 -4.120 -17.417 6.734 1.00 22.03 ATOM 59 CA GLU 164 -3.033 -16.634 7.305 1.00 22.03 ATOM 60 CB GLU 164 -1.875 -17.541 7.725 1.00 17.15 ATOM 61 CG GLU 164 -2.198 -18.414 8.937 1.00 17.15 ATOM 62 CD GLU 164 -1.114 -19.434 9.249 1.00 17.15 ATOM 63 OE1 GLU 164 -0.283 -19.710 8.3611.00 17.15 ATOM 64 OE2 GLU 164 -1.099 -19.968 10.379 1.00 17.15 ATOM 65 C GLU 164 -2.559 -15.542 6.354 1.00 22.03 ATOM 66 O GLU 164 -2.160 -14.470 6.802 1.00 17.15 ATOM 67 N TRP 165 -2.607 -15.805 5.048 1.00 10.72 ATOM 68 CA TRP 165 -2.205-14.803 4.063 1.00 10.72 ATOM 69 CB TRP 165 -2.223 -15.377 2.644 1.00 2.00 ATOM 70 CG TRP 165 -0.928 -16.003 2.227 1.00 2.00 ATOM 71 CD2 TRP 165 0.350 -15.358 2.131 1.00 2.00 ATOM 72 CE2 TRP 165 1.275 -16.326 1.685 1.00 2.00 ATOM 73 CE3 TRP 1650.804 -14.054 2.379 1.00 2.00 ATOM 74 CD1 TRP 165 -0.731 -17.298 1.848 1.00 2.00 ATOM 75 NE1 TRP 165 0.587 -17.500 1.521 1.00 2.00 ATOM 76 CZ2 TRP 165 2.627 -16.036 1.479 1.00 2.00 ATOM 77 CZ3 TRP 165 2.152 -13.764 2.174 1.00 2.00 ATOM 78 CH2TRP 165 3.046 -14.754 1.729 1.00 2.00 ATOM 79 C TRP 165 -3.137 -13.601 4.149 1.00 10.72 ATOM 80 O TRP 165 -2.717 -12.463 3.925 1.00 2.00 ATOM 81 N ASP 166 -4.408 -13.861 4.441 1.00 14.80 ATOM 82 CA ASP 166 -5.397 -12.796 4.580 1.00 14.80 ATOM83 CB ASP 166 -6.812 -13.370 4.698 1.00 28.74 ATOM 84 CG ASP 166 -7.298 -13.999 3.403 1.00 28.74 ATOM 85 OD1 ASP 166 -6.909 -13.511 2.320 1.00 28.74 ATOM 86 OD2 ASP 166 -8.071 -14.978 3.466 1.00 28.74 ATOM 87 C ASP 166 -5.063 -11.981 5.819 1.00 14.80 ATOM 88 O ASP 166 -5.056 -10.749 5.775 1.00 28.74 ATOM 89 N LEU 167 -4.745 -12.682 6.906 1.00 11.01 ATOM 90 CA LEU 167 -4.383 -12.044 8.166 1.00 11.01 ATOM 91 CB LEU 167 -4.036 -13.103 9.214 1.00 31.53 ATOM 92 CG LEU 167 -4.672 -12.97510.601 1.00 31.53 ATOM 93 CD1 LEU 167 -3.806 -13.709 11.619 1.00 31.53 ATOM 94 CD2 LEU 167 -4.820 -11.507 10.989 1.00 31.53 ATOM 95 C LEU 167 -3.161 -11.159 7.933 1.00 11.01 ATOM 96 O LEU 167 -3.120 -10.006 8.367 1.00 31.53 ATOM 97 N ILE 168-2.180 -11.714 7.228 1.00 13.18 ATOM 98 CA ILE 168 -0.937 -11.027 6.900 1.00 13.18 ATOM 99 CB ILE 168 0.015 -11.968 6.113 1.00 18.30 ATOM 100 CG2 ILE 168 1.118 -11.182 5.414 1.00 18.30 ATOM 101 CG1 ILE 168 0.604 -13.013 7.063 1.00 18.30 ATOM102 CD1 ILE 168 1.379 -14.111 6.373 1.00 18.30 ATOM 103 C ILE 168 -1.185 -9.747 6.107 1.00 13.18 ATOM 104 O ILE 168 -0.637 -8.697 6.437 1.00 18.30

ATOM 105 N HIS 169 -2.032 -9.831 5.084 1.00 12.99 ATOM 106 CA HIS 169 -2.342 -8.674 4.245 1.00 12.99 ATOM 107 CB HIS 169 -3.218 -9.087 3.062 1.00 13.09 ATOM 108 CG HIS 169 -2.553 -10.045 2.126 1.00 13.09 ATOM 109 CD2 HIS 169 -1.247-10.223 1.811 1.00 13.09 ATOM 110 ND1 HIS 169 -3.249 -11.000 1.416 1.00 13.09 ATOM 111 CE1 HIS 169 -2.403 -11.728 0.710 1.00 13.09 ATOM 112 NE2 HIS 169 -1.181 -11.277 0.936 1.00 13.09 ATOM 113 C HIS 169 -3.017 -7.550 5.017 1.00 12.99 ATOM 114 OHIS 169 -2.680 -6.377 4.839 1.00 13.09 ATOM 115 N VAL 170 -3.978 -7.909 5.862 1.00 13.36 ATOM 116 CA VAL 170 -4.696 -6.926 6.664 1.00 13.36 ATOM 117 CB VAL 110 -5.863 -7.572 7.443 1.00 20.12 ATOM 118 CG1 VAL 170 -6.541 -6.540 8.340 1.00 20.12 ATOM 119 CG2 VAL 170 -6.869 -8.165 6.471 1.00 20.12 ATOM 120 C VAL 170 -3.741 -6.246 7.639 1.00 13.36 ATOM 121 O VAL 170 -3.728 -5.019 7.744 1.00 20.12 ATOM 122 N ALA 171 -2.920 -7.043 8.320 1.00 11.04 ATOM 123 CA ALA 171 -1.953 -6.515 9.277 1.00 11.04 ATOM 124 CB ALA 171 -1.249 -7.653 10.005 1.00 13.43 ATOM 125 C ALA 171 -0.931 -5.613 8.588 1.00 11.04 ATOM 126 O ALA 171 -0.658 -4.507 9.058 1.00 13.43 ATOM 127 N THR 172 -0.382 -6.076 7.469 1.00 12.51 ATOM 128 CA THR 172 0.606 -5.3016.723 1.00 12.51 ATOM 129 CB THR 172 1.062 -6.032 5.445 1.00 14.17 ATOM 130 OG1 THR 172 1.548 -7.338 5.782 1.00 14.17 ATOM 131 CG2 THR 172 2.175 -5.255 4.756 1.00 14.17 ATOM 132 C THR 172 0.045 -3.936 6.337 1.00 12.51 ATOM 133 O THR 172 0.701-2.910 6.537 1.00 14.17 ATOM 134 N GLU 173 -1.178 -3.921 5.815 1.00 17.79 ATOM 135 CA GLU 173 -1.818 -2.675 5.421 1.00 17.79 ATOM 136 CB GLU 173 -3.130 -2.946 4.682 1.00 49.44 ATOM 137 CG GLU 173 -3.823 -1.679 4.171 1.00 49.44 ATOM 138 CD GLU173 -2.930 -0.835 3.266 1.00 49.44 ATOM 139 OE1 GLU 173 -2.075 -1.408 2.552 1.00 49.44 ATOM 140 OE2 GLU 173 -3.085 0.404 3.269 1.00 49.44 ATOM 141 C GLU 173 -2.072 -1.780 6.628 1.00 17.79 ATOM 142 O GLU 173 -1.854 -0.568 6.557 1.00 49.44 ATOM143 N ALA 174 -2.525 -2.375 7.731 1.00 13.12 ATOM 144 CA ALA 174 -2.798 -1.631 8.957 1.00 13.12 ATOM 145 CB ALA 174 -3.226 -2.576 10.068 1.00 17.51 ATOM 146 C ALA 174 -1.556 -0.856 9.375 1.00 13.12 ATOM 147 O ALA 174 -1.634 0.319 9.735 1.00 17.51 ATOM 148 N HIS 175 -0.409 -1.521 9.317 1.00 12.20 ATOM 149 CA HIS 175 0.851 -0.895 9.679 1.00 12.20 ATOM 150 CB HIS 175 1.944 -1.949 9.886 1.00 17.52 ATOM 151 CG HIS 175 3.302 -1.365 10.136 1.00 17.52 ATOM 152 CD2 HIS 175 3.733 -0.46811.055 1.00 17.52 ATOM 153 ND1 HIS 175 4.400 -1.679 9.364 1.00 17.52 ATOM 154 CE1 HIS 175 5.447 -0.999 9.793 1.00 17.52 ATOM 155 NE2 HIS 175 5.070 -0.258 10.818 1.00 17.52 ATOM 156 C HIS 175 1.311 0.133 8.654 1.00 12.20 ATOM 157 O HIS 175 1.7001.240 9.024 1.00 17.52 ATOM 158 N ARG 176 1.291 -0.233 7.375 1.00 12.54 ATOM 159 CA ARG 176 1.735 0.677 6.328 1.00 12.54 ATOM 160 CB ARG 176 1.662 0.017 4.950 1.00 50.41 ATOM 161 CG ARG 176 2.683 -1.088 4.730 1.00 50.41 ATOM 162 CD ARG 1762.666 -1.565 3.299 1.00 50.41 ATOM 163 NE ARG 176 3.682 -2.571 2.989 1.00 50.41 ATOM 164 CZ ARG 176 3.577 -3.472 2.012 1.00 50.41 ATOM 165 NH1 ARG 176 2.496 -3.513 1.236 1.00 50.41 ATOM 166 NH2 ARG 176 4.536 -4.376 1.841 1.00 50.41 ATOM 167 CARG 176 0.972 1.988 6.306 1.00 12.54 ATOM 168 O ARG 176 1.561 3.040 6.087 1.00 50.41 ATOM 169 N SER 177 -0.326 1.935 6.581 1.00 24.74 ATOM 170 CA SER 177 -1.147 3.145 6.584 1.00 24.74 ATOM 171 CB SER 177 -2.622 2.792 6.414 1.00 21.56 ATOM 172OG SER 177 -3.069 1.913 7.436 1.00 21.56 ATOM 173 C SER 177 -0.960 4.013 7.832 1.00 24.74 ATOM 174 O SER 177 -1.401 5.159 7.863 1.00 21.56 ATOM 175 N THR 178 -0.347 3.453 8.870 1.00 17.96 ATOM 176 CA THR 178 -0.104 4.181 10.115 1.00 17.96 ATOM177 CB THR 178 -0.736 3.440 11.323 1.00 19.76 ATOM 178 OG1 THR 178 -0.265 2.091 11.361 1.00 19.76 ATOM 179 CG2 THR 178 -2.253 3.443 11.211 1.00 19.76 ATOM 180 C THR 178 1.376 4.395 10.382 1.00 17.96 ATOM 181 O THR 178 1.760 4.880 11.445 1.00 19.76 ATOM 182 N ASN 179 2.207 4.024 9.417 1.00 25.88 ATOM 183 CA ASN 179 3.654 4.180 9.546 1.00 25.88 ATOM 184 CB ASN 179 4.362 2.974 8.943 1.00 44.29 ATOM 185 CG ASN 179 5.817 2.871 9.368 1.00 44.29 ATOM 186 OD1 ASN 179 6.129 2.768 10.564 1.00 44.29 ATOM 187 ND2 ASN 179 6.719 2.830 8.391 1.00 44.29 ATOM 188 C ASN 179 4.078 5.458 8.823 1.00 25.88 ATOM 189 O ASN 179 4.150 5.495 7.590 1.00 44.29 ATOM 190 N ALA 180 4.332 6.502 9.604 1.00 45.20 ATOM 191 CA ALA 180 4.740 7.818 9.126 1.00 45.20 ATOM 192 CB ALA 180 5.026 8.743 10.313 1.00 36.14 ATOM 193 C ALA 180 5.931 7.808 8.170 1.00 45.20 ATOM 194 O ALA 180 6.918 7.097 8.372 1.00 36.14 ATOM 195 N ALA 181 5.784 8.552 7.086 1.00 44.05 ATOM 196 CA ALA 181 6.834 8.661 6.072 1.00 44.05 ATOM 197 CB ALA 181 8.170 9.116 6.722 1.00 50.21 ATOM 198 C ALA 181 7.069 7.427 5.196 1.00 44.05 ATOM 199 O ALA 181 7.663 7.550 4.118 1.00 50.21 ATOM 200 N GLY 182 6.567 6.268 5.622 1.00 39.06 ATOM 201 CA GLY 182 6.756 5.040 4.867 1.00 39.06 ATOM 202 C GLY 182 8.202 4.769 4.482 1.00 39.06 ATOM 203 O GLY 182 9.096 4.785 5.334 1.00 48.58 ATOM 204 N SER 183 8.438 4.564 3.189 1.00 64.55 ATOM 205 CA SER 183 9.781 4.270 2.693 1.00 64.55 ATOM 206 CB SER 183 9.690 3.402 1.430 1.00 67.68 ATOM 207 OG SER 183 8.822 3.978 0.467 1.00 67.68 ATOM 208 C SER 183 10.643 5.510 2.437 1.00 64.55 ATOM 209 O SER 183 11.839 5.407 2.158 1.00 67.68 ATOM 210 N HIS 184 10.035 6.683 2.579 1.00 52.73 ATOM 211 CA HIS 184 10.725 7.953 2.352 1.00 52.73 ATOM 212 CB HIS 184 9.772 8.955 1.698 1.00 44.77 ATOM 213 C HIS 184 11.364 8.582 3.595 1.00 52.73 ATOM 214 O HIS 184 11.837 9.722 3.540 1.00 44.77 ATOM 215 N TRP 185 11.420 7.842 4.699 1.00 54.14 ATOM 216 CA TRP 185 11.977 8.389 5.940 1.00 54.14 ATOM 217 CB TRP 185 11.813 7.395 7.104 1.00 40.24 ATOM 218 CG TRP 185 12.605 6.123 6.991 1.00 40.24 ATOM 219 CD2 TRP 185 13.894 5.873 7.551 1.00 40.24 ATOM 220 CE2 TRP 185 14.245 4.543 7.221 1.00 40.24 ATOM 221 CE3 TRP 185 14.791 6.6418.300 1.00 40.24 ATOM 222 CD1 TRP 185 12.227 4.973 6.359 1.00 40.24 ATOM 223 NE1 TRP 185 13.210 4.015 6.496 1.00 40.24 ATOM 224 CZ2 TRP 185 15.461 3.968 7.619 1.00 40.24 ATOM 225 CZ3 TRP 185 15.996 6.073 8.696 1.00 40.24 ATOM 226 CH2 TRP 18516.319 4.747 8.353 1.00 40.24 ATOM 227 C TRP 185 13.432 8.870 5.819 1.00 54.14 ATOM 228 O TRP 185 13.759 10.008 6.168 1.00 40.24 ATOM 229 N LYS 186 14.277 8.032 5.232 1.00 43.72 ATOM 230 CA LYS 186 15.694 8.329 5.035 1.00

43.72 ATOM 231 CB LYS 186 16.353 7.168 4.282 1.00 64.14 ATOM 232 CG LYS 186 17.830 7.355 3.945 1.00 64.14 ATOM 233 CD LYS 186 18.758 7.175 5.139 1.00 64.14 ATOM 234 CE LYS 186 20.195 7.060 4.652 1.00 64.14 ATOM 235 NZ LYS 186 20.3485.838 3.805 1.00 64.14 ATOM 236 C LYS 186 15.900 9.634 4.263 1.00 43.72 ATOM 237 O LYS 186 16.948 10.256 4.366 1.00 64.14 ATOM 238 N GLN 187 14.892 10.032 3.491 1.00 58.06 ATOM 239 CA GLN 187 14.958 11.244 2.682 1.00 58.06 ATOM 240 CB GLN 18714.288 10.997 1.321 1.00 74.68 ATOM 241 CG GLN 187 14.639 9.662 0.667 1.00 74.68 ATOM 242 CD GLN 187 16.133 9.397 0.607 1.00 74.68 ATOM 243 OE1 GLN 187 16.926 10.312 0.381 1.00 74.68 ATOM 244 NE2 GLN 187 16.528 8.156 0.855 1.00 74.68 ATOM 245 CGLN 187 14.322 12.466 3.342 1.00 58.06 ATOM 246 O GLN 187 14.897 13.551 3.358 1.00 74.68 ATOM 247 N ARG 188 13.117 12.280 3.866 1.00 54.11 ATOM 248 CA ARG 188 12.363 13.360 4.505 1.00 54.11 ATOM 249 CB ARG 188 10.889 13.115 4.334 1.00 53.33 ATOM 250 C ARG 188 12.654 13.626 5.977 1.00 54.11 ATOM 251 O ARG 188 11.879 14.298 6.659 1.00 53.33 ATOM 252 N ARG 189 13.754 13.090 6.473 1.00 39.52 ATOM 253 CA ARG 189 14.089 13.271 7.875 1.00 39.52 ATOM 254 CB ARG 189 14.594 11.959 8.482 1.00 60.85 ATOM 255 CG ARG 189 15.969 11.555 7.991 1.00 60.85 ATOM 256 CD ARG 189 16.442 10.298 8.693 1.00 60.85 ATOM 257 NE ARG 189 17.833 9.963 8.385 1.00 60.85 ATOM 258 CZ ARG 189 18.627 9.261 9.190 1.00 60.85 ATOM 259 NH1 ARG 189 18.178 8.80510.356 1.00 60.85 ATOM 260 NH2 ARG 189 19.882 9.021 8.841 1.00 60.85 ATOM 261 C ARG 189 15.109 14.378 8.109 1.00 39.52 ATOM 262 O ARG 189 16.037 14.565 7.320 1.00 60.85 ATOM 263 N LYS 190 14.934 15.100 9.212 1.00 44.13 ATOM 264 CA LYS 19015.834 16.183 9.586 1.00 44.13 ATOM 265 CB LYS 190 15.068 17.500 9.680 1.00 45.33 ATOM 266 C LYS 190 16.472 15.846 10.928 1.00 44.13 ATOM 267 O LYS 190 15.827 15.272 11.805 1.00 45.33 ATOM 268 N PHE 191 17.748 16.184 11.067 1.00 35.64 ATOM 269CA PHE 191 18.489 15.928 12.291 1.00 35.64 ATOM 270 CB PHE 191 19.993 16.008 12.025 1.00 53.94 ATOM 271 CG PHE 191 20.550 14.827 11.286 1.00 53.94 ATOM 272 CD1 PHE 191 20.209 14.596 9.958 1.00 53.94 ATOM 273 CD2 PHE 191 21.430 13.949 11.915 1.00 53.94 ATOM 274 CE1 PHE 191 20.735 13.510 9.265 1.00 53.94 ATOM 275 CE2 PHE 191 21.964 12.859 11.230 1.00 53.94 ATOM 276 CZ PHE 191 21.615 12.639 9.900 1.00 53.94 ATOM 277 C PHE 191 18.135 16.928 13.384 1.00 35.64 ATOM 278 O PHE 191 17.997 18.12713.120 1.00 53.94 ATOM 279 N LEU 192 17.978 16.439 14.610 1.00 44.53 ATOM 280 CA LEU 192 17.683 17.315 15.736 1.00 44.53 ATOM 281 CB LEU 192 17.326 16.493 16.980 1.00 22.94 ATOM 282 CG LEU 192 16.931 17.259 18.246 1.00 22.94 ATOM 283 CD1 LEU192 15.568 17.906 18.064 1.00 22.94 ATOM 284 CD2 LEU 192 16.909 16.308 19.427 1.00 22.94 ATOM 285 C LEU 192 18.974 18.101 15.980 1.00 44.53 ATOM 286 O LEU 192 20.049 17.507 16.129 1.00 22.94 ATOM 287 N PRO 193 18.895 19.444 15.977 1.00 34.26 ATOM 288 CD PRO 193 17.670 20.241 15.781 1.00 46.23 ATOM 289 CA PRO 193 20.058 20.311 16.198 1.00 34.26 ATOM 290 CB PRO 193 19.417 21.670 16.465 1.00 46.23 ATOM 291 CG PRO 193 18.213 21.641 15.579 1.00 46.23 ATOM 292 C PRO 193 20.917 19.84417.372 1.00 34.26 ATOM 293 O PRO 193 20.413 19.614 18.471 1.00 46.23 ATOM 294 N ASP 194 22.217 19.716 17.125 1.00 42.67 ATOM 295 CA ASP 194 23.174 19.254 18.128 1.00 42.67 ATOM 296 CB ASP 194 24.583 19.226 17.536 1.00 68.50 ATOM 297 CG ASP 19424.731 18.185 16.450 1.00 68.50 ATOM 298 OD1 ASP 194 25.066 17.027 16.782 1.00 68.50 ATOM 299 OD2 ASP 194 24.498 18.518 15.269 1.00 68.50 ATOM 300 C ASP 194 23.187 20.003 19.457 1.00 42.67 ATOM 301 O ASP 194 23.545 19.432 20.486 1.00 68.50 ATOM302 N ASP 195 22.817 21.280 19.438 1.00 47.52 ATOM 303 CA ASP 195 22.793 22.070 20.666 1.00 47.52 ATOM 304 CB ASP 195 22.586 23.559 20.351 1.00 85.02 ATOM 305 CG ASP 195 21.327 23.824 19.537 1.00 85.02 ATOM 306 OD1 ASP 195 20.291 24.188 20.1381.00 85.02 ATOM 307 OD2 ASP 195 21.377 23.683 18.294 1.00 85.02 ATOM 308 C ASP 195 21.715 21.561 21.627 1.00 47.52 ATOM 309 O ASP 195 21.762 21.826 22.831 1.00 85.02 ATOM 310 N ILE 196 20.760 20.810 21.089 1.00 44.54 ATOM 311 CA ILE 196 19.66320.259 21.875 1.00 44.54 ATOM 312 CB ILE 196 18.379 20.137 21.023 1.00 39.66 ATOM 313 CG2 ILE 196 17.223 19.627 21.874 1.00 39.66 ATOM 314 CG1 ILE 196 18.031 21.496 20.407 1.00 39.66 ATOM 315 CD1 ILE 196 16.816 21.475 19.503 1.00 39.66 ATOM 316C ILE 196 20.030 18.882 22.420 1.00 44.54 ATOM 317 O ILE 196 20.582 18.046 21.705 1.00 39.66 ATOM 318 N GLY 197 19.714 18.652 23.690 1.00 42.85 ATOM 319 CA GLY 1.97 20.006 17.372 24.307 1.00 42.85 ATOM 320 C GLY 197 21.371 17.285 24.956 1.00 42.85 ATOM 321 O GLY 197 21.815 16.198 25.318 1.00 40.22 ATOM 322 N GLN 198 22.029 18.425 25.137 1.00 53.07 ATOM 323 CA GLN 198 23.351 18.444 25.754 1.00 53.07 ATOM 324 CB GLN 198 24.357 19.103 24.810 1.00 44.23 ATOM 325 C GLN 198 23.344 19.15327.110 1.00 53.07 ATOM 326 O GLN 198 24.396 19.545 27.616 1.00 44.23 ATOM 327 N SER 199 22.170 19.244 27.729 1.00 35.30 ATOM 328 CA SER 199 22.037 19.918 29.019 1.00 35.30 ATOM 329 CB SER 199 21.472 21.328 28.806 1.00 58.72 ATOM 330 OG SER 19922.093 21.971 27.704 1.00 58.72 ATOM 331 C SER 199 21.168 19.169 30.036 1.00 35.30 ATOM 332 O SER 199 20.135 19.681 30.482 1.00 58.72 ATOM 333 N PRO 200 21.544 17.928 30.387 1.00 34.70 ATOM 334 CD PRO 200 22.656 17.108 29.872 1.00 38.71 ATOM335 CA PRO 200 20.740 17.184 31.362 1.00 34.70 ATOM 336 CB PRO 200 21.311 15.769 31.266 1.00 38.71 ATOM 337 CG PRO 200 22.737 15.992 30.878 1.00 38.71 ATOM 338 C PRO 200 20.923 17.784 32.759 1.00 34.70 ATOM 339 O PRO 200 22.006 17.692 33.341 1.00 38.71 ATOM 340 N ILE 201 19.876 18.413 33.286 1.00 42.94 ATOM 341 CA ILE 201 19.961 19.041 34.604 1.00 42.94 ATOM 342 CB ILE 201 20.059 20.582 34.491 1.00 51.32 ATOM 343 CG2 ILE 201 21.468 20.991 34.078 1.00 51.32 ATOM 344 CG1 ILE 201 19.00921.111 33.510 1.00 51.32 ATOM 345 CD1 ILE 201 19.169 22.582 33.164 1.00 51.32 ATOM 346 C ILE 201 18.871 18.676 35.610 1.00 42.94 ATOM 347 O ILE 201 19.049 18.875 36.814 1.00 51.32 ATOM 348 N VAL 202 17.737 18.172 35.133 1.00 50.33 ATOM 349 CAVAL 202 16.661 17.787 36.043 1.00 50.33 ATOM 350 CB VAL 202 15.296 17.722 35.326 1.00 36.59 ATOM 351 CG1 VAL 202 14.202 17.311 36.304 1.00 36.59 ATOM 352 CG2 VAL 202 14.968 19.074 34.714 1.00 36.59 ATOM 353 C VAL 202 17.007 16.435 36.665 1.00 50.33 ATOM 354 O VAL 202 17.335 15.481 35.955 1.00 36.59 ATOM 355 N SER 203 16.960 16.375 37.991 1.00 49.46

ATOM 356 CA SER 203 17.289 15.166 38.736 1.00 49.46 ATOM 357 CB SER 203 17.298 15.467 40.241 1.00 64.20 ATOM 358 OG SER 203 17.673 14.330 41.003 1.00 64.20 ATOM 359 C SER 203 16.356 13.992 38.463 1.00 49.46 ATOM 360 O SER 203 15.14714.166 38.310 1.00 64.20 ATOM 361 N MET 204 16.944 12.800 38.419 1.00 41.99 ATOM 362 CA MET 204 16.223 11.551 38.205 1.00 41.99 ATOM 363 CB MET 204 16.320 11.096 36.746 1.00 48.64 ATOM 364 CG MET 204 15.470 11.895 35.771 1.00 48.64 ATOM 365 SDMET 204 13.702 11.783 36.114 1.00 48.64 ATOM 366 CE MET 204 13.284 10.257 35.264 1.00 48.64 ATOM 367 C MET 204 16.900 10.528 39.109 1.00 41.99 ATOM 368 O MET 204 18.127 10.417 39.121 1.00 48.64 ATOM 369 N PRO 205 16.108 9.754 39.869 1.00 38.42 ATOM 370 CD PRO 205 14.633 9.815 39.866 1.00 52.20 ATOM 371 CA PRO 205 16.586 8.724 40.797 1.00 38.42 ATOM 372 CB PRO 205 15.334 7.888 41.041 1.00 52.20 ATOM 373 CG PRO 205 14.254 8.919 41.028 1.00 52.20 ATOM 374 C PRO 205 17.769 7.858 40.3401.00 38.42 ATOM 375 O PRO 205 18.724 7.675 41.092 1.00 52.20 ATOM 376 N ASP 206 17.720 7.349 39.111 1.00 49.06 ATOM 377 CA ASP 206 18.791 6.490 38.601 1.00 49.06 ATOM 378 CB ASP 206 18.282 5.627 37.437 1.00 74.42 ATOM 379 CG ASP 206 17.6906.450 36.305 1.00 74.42 ATOM 380 OD1 ASP 206 18.397 7.335 35.770 1.00 74.42 ATOM 381 OD2 ASP 206 16.516 6.199 35.948 1.00 74.42 ATOM 382 C ASP 206 20.106 7.177 38.214 1.00 49.06 ATOM 383 O ASP 206 21.069 6.506 37.838 1.00 74.42 ATOM 384 N GLY207 20.139 8.505 38.272 1.00 42.48 ATOM 385 CA GLY 207 21.355 9.225 37.928 1.00 42.48 ATOM 386 C GLY 207 21.330 9.965 36.601 1.00 42.48 ATOM 387 O GLY 207 21.890 11.058 36.494 1.00 42.50 ATOM 388 N ASP 208 20.725 9.365 35.581 1.00 46.70 ATOM389 CA ASP 208 20.636 9.999 34.266 1.00 46.70 ATOM 390 CB ASP 208 20.162 8.994 33.212 1.00 61.56 ATOM 391 CG ASP 208 21.143 7.856 33.006 1.00 61.56 ATOM 392 OD1 ASP 208 20.723 6.684 33.122 1.00 61.56 ATOM 393 OD2 ASP 208 22.330 8.134 32.724 1.00 61.56 ATOM 394 C ASP 208 19.666 11.176 34.339 1.00 46.70 ATOM 395 O ASP 208 18.462 10.983 34.506 1.00 61.56 ATOM 396 N LYS 209 20.200 12.389 34.238 1.00 41.30 ATOM 397 CA LYS 209 19.389 13.602 34.308 1.00 41.30 ATOM 398 CB LYS 209 20.254 14.78234.732 1.00 41.38 ATOM 399 C LYS 209 18.657 13.916 33.004 1.00 41.30 ATOM 400 O LYS 209 19.052 13.458 31.930 1.00 41.38 ATOM 401 N VAL 210 17.603 14.723 33.109 1.00 43.36 ATOM 402 CA VAL 210 16.792 15.107 31.954 1.00 43.36 ATOM 403 CB VAL 21015.275 15.014 32.282 1.00 30.23 ATOM 404 CG1 VAL 210 14.440 15.358 31.055 1.00 30.23 ATOM 405 CG2 VAL 210 14.923 13.624 32.782 1.00 30.23 ATOM 406 C VAL 210 17.088 16.522 31.442 1.00 43.36 ATOM 407 O VAL 210 17.395 17.430 32.221 1.00 30.23 ATOM408 N ASP 211 17.004 16.685 30.125 1.00 27.49 ATOM 409 CA ASP 211 17.217 17.966 29.458 1.00 27.49 ATOM 410 CB ASP 211 18.073 17.765 28.198 1.00 30.75 ATOM 411 CG ASP 211 18.360 19.068 27.447 1.00 30.75 ATOM 412 OD1 ASP 211 19.473 19.196 26.9001.00 30.75 ATOM 413 OD2 ASP 211 17.484 19.955 27.370 1.00 30.75 ATOM 414 C ASP 211 15.819 18.445 29.073 1.00 27.49 ATOM 415 O ASP 211 15.197 17.892 28.166 1.00 30.75 ATOM 416 N LEU 212 15.343 19.488 29.745 1.00 31.99 ATOM 417 CA LEU 212 14.01320.042 29.492 1.00 31.99 ATOM 418 CB LEU 212 13.778 21.274 30.369 1.00 35.19 ATOM 419 CG LEU 212 13.606 20.997 31.864 1.00 35.19 ATOM 420 CD1 LEU 212 13.621 22.298 32.652 1.00 35.19 ATOM 421 CD2 LEU 212 12.309 20.237 32.098 1.00 35.19 ATOM 422C LEU 212 13.713 20.377 28.032 1.00 31.99 ATOM 423 O LEU 212 12.625 20.083 27.539 1.00 35.19 ATOM 424 N GLU 213 14.672 20.981 27.338 1.00 28.70 ATOM 425 CA GLU 213 14.468 21.345 25.940 1.00 28.70 ATOM 426 CB GLU 213 15.623 22.209 25.428 1.00 62.21 ATOM 427 CG GLU 213 15.434 22.707 23.997 1.00 62.21 ATOM 428 CD GLU 213 16.651 23.440 23.446 1.00 62.21 ATOM 429 OE1 GLU 213 17.778 23.214 23.945 1.00 62.21 ATOM 430 OE2 GLU 213 16.478 24.237 22.498 1.00 62.21 ATOM 431 C GLU 213 14.31720.104 25.067 1.00 28.70 ATOM 432 O GLU 213 13.403 20.024 24.247 1.00 62.21 ATOM 433 N ALA 214 15.201 19.130 25.262 1.00 28.17 ATOM 434 CA ALA 214 15.162 17.890 24.494 1.00 28.17 ATOM 435 CB ALA 214 16.330 16.998 24.872 1.00 42.74 ATOM 436 CALA 214 13.844 17.176 24.759 1.00 28.17 ATOM 437 O ALA 214 13.174 16.726 23.829 1.00 42.74 ATOM 438 N PHE 215 13.468 17.104 26.032 1.00 21.66 ATOM 439 CA PHE 215 12.222 16.471 26.444 1.00 21.66 ATOM 440 CB PHE 215 12.033 16.628 27.958 1.00 28.76 ATOM 441 CG PHE 215 10.751 16.038 28.481 1.00 28.76 ATOM 442 CD1 PHE 215 10.675 14.689 28.815 1.00 28.76 ATOM 443 CD2 PHE 215 9.623 16.835 28.653 1.00 28.76 ATOM 444 CE1 PHE 215 9.493 14.143 29.315 1.00 28.76 ATOM 445 CE2 PHE 215 8.438 16.30029.150 1.00 28.76 ATOM 446 CZ PHE 215 8.373 14.951 29.482 1.00 28.76 ATOM 447 C PHE 215 11.068 17.132 25.696 1.00 21.66 ATOM 448 O PHE 215 10.215 16.451 25.122 1.00 28.76 ATOM 449 N SER 216 11.073 18.462 25.680 1.00 28.03 ATOM 450 CA SER 21610.043 19.242 25.007 1.00 28.03 ATOM 451 CB SER 216 10.349 20.734 25.146 1.00 33.85 ATOM 452 OG SER 216 9.300 21.529 24.624 1.00 33.85 ATOM 453 C SER 216 9.945 18.857 23.532 1.00 28.03 ATOM 454 O SER 216 8.852 18.613 23.019 1.00 33.85 ATOM 455N GLU 217 11.092 18.761 22.868 1.00 28.84 ATOM 456 CA GLU 217 11.138 18.402 21.454 1.00 28.84 ATOM 457 CB GLU 217 12.581 18.420 20.943 1.00 47.68 ATOM 458 CG GLU 217 13.174 19.815 20.811 1.00 47.68 ATOM 459 CD GLU 217 12.405 20.684 19.829 1.00 47.68 ATOM 460 OE1 GLU 217 11.660 21.581 20.281 1.00 47.68 ATOM 461 OE2 GLU 217 12.542 20.465 18.606 1.00 47.68 ATOM 462 C GLU 217 10.505 17.044 21.179 1.00 28.84 ATOM 463 O GLU 217 9.751 16.886 20.217 1.00 47.68 ATOM 464 N PHE 218 10.799 16.07122.036 1.00 21.49 ATOM 465 CA PHE 218 10.259 14.725 21.883 1.00 21.49 ATOM 466 CB PHE 218 11.020 13.746 22.781 1.00 24.12 ATOM 467 CG PHE 218 12.489 13.652 22.464 1.00 24.12 ATOM 468 CD1 PHE 218 13.431 13.554 23.481 1.00 24.12 ATOM 469 CD2 PHE218 12.932 13.677 21.144 1.00 24.12 ATOM 470 CE1 PHE 218 14.793 13.484 23.187 1.00 24.12 ATOM 471 CE2 PHE 218 14.290 13.607 20.843 1.00 24.12 ATOM 472 CZ PHE 218 15.221 13.511 21.867 1.00 24.12 ATOM 473 C PHE 218 8.765 14.675 22.176 1.00 21.49 ATOM 474 O PHE 218 7.985 14.166 21.369 1.00 24.12 ATOM 475 N THR 219 8.358 15.227 23.312 1.00 20.07 ATOM 476 CA THR 219 6.949 15.231 23.685 1.00 20.07 ATOM 477 CB THR 219 6.741 15.766 25.118 1.00 28.98 ATOM 478 OG1 THR 219 7.418 17.021 25.2741.00 28.98 ATOM 479 CG2 THR 219 7.275 14.767 26.132 1.00 28.98 ATOM 480 C THR 219 6.080 16.011 22.696 1.00 20.07 ATOM 481 O THR 219 4.914 15.670 22.482 1.00

28.98 ATOM 482 N LYS 220 6.662 17.022 22.060 1.00 25.35 ATOM 483 CA LYS 220 5.943 17.840 21.088 1.00 25.35 ATOM 484 CB LYS 220 6.842 18.965 20.577 1.00 29.07 ATOM 485 C LYS 220 5.414 17.015 19.916 1.00 25.35 ATOM 486 O LYS 220 4.37617.343 19.339 1.00 29.07 ATOM 487 N ILE 221 6.122 15.943 19.569 1.00 31.43 ATOM 488 CA ILE 221 5.708 15.089 18.458 1.00 31.43 ATOM 489 CB ILE 221 6.842 14.915 17.413 1.00 25.19 ATOM 490 CG2 ILE 221 7.240 16.264 16.838 1.00 25.19 ATOM 491 CG1ILE 221 8.050 14.215 18.043 1.00 25.19 ATOM 492 CD1 ILE 221 9.113 13.799 17.044 1.00 25.19 ATOM 493 C ILE 221 5.240 13.700 18.892 1.00 31.43 ATOM 494 O ILE 221 4.930 12.857 18.046 1.00 25.19 ATOM 495 N ILE 222 5.129 13.474 20.198 1.00 24.41 ATOM 496 CA ILE 222 4.720 12.162 20.687 1.00 24.41 ATOM 497 CB ILE 222 5.189 11.916 22.147 1.00 27.10 ATOM 498 CG2 ILE 222 4.221 12.545 23.145 1.00 27.10 ATOM 499 CG1 ILE 222 5.302 10.410 22.400 1.00 27.10 ATOM 500 CD1 ILE 222 6.062 10.053 23.6461.00 27.10 ATOM 501 C ILE 222 3.231 11.845 20.541 1.00 24.41 ATOM 502 O ILE 222 2.864 10.691 20.307 1.00 27.10 ATOM 503 N THR 223 2.378 12.861 20.642 1.00 33.16 ATOM 504 CA THR 223 0.936 12.653 20.520 1.00 33.16 ATOM 505 CB THR 223 0.150 13.97420.721 1.00 36.84 ATOM 506 OG1 THR 223 0.352 14.442 22.063 1.00 36.84 ATOM 507 CG2 THR 223 -1.346 13.764 20.484 1.00 36.84 ATOM 508 C THR 223 0.536 11.954 19.212 1.00 33.16 ATOM 509 O THR 223 -0.156 10.932 19.242 1.00 36.84 ATOM 510 N PRO 2240.968 12.482 18.048 1.00 18.75 ATOM 511 CD PRO 224 1.691 13.735 17.770 1.00 26.12 ATOM 512 CA PRO 224 0.590 11.805 16.802 1.00 18.75 ATOM 513 CB PRO 224 1.117 12.747 15.715 1.00 26.12 ATOM 514 CG PRO 224 2.221 13.497 16.386 1.00 26.12 ATOM 515C PRO 224 1.200 10.402 16.701 1.00 18.75 ATOM 516 O PRO 224 0.606 9.502 16.101 1.00 26.12 ATOM 517 N ALA 225 2.368 10.213 17.312 1.00 12.19 ATOM 518 CA ALA 225 3.040 8.916 17.300 1.00 12.19 ATOM 519 CB ALA 225 4.415 9.021 17.943 1.00 20.39 ATOM520 C ALA 225 2.187 7.881 18.030 1.00 12.19 ATOM 521 O ALA 225 1.998 6.764 17.545 1.00 20.39 ATOM 522 N ILE 226 1.645 8.271 19.179 1.00 14.61 ATOM 523 CA ILE 226 0.798 7.385 19.971 1.00 14.61 ATOM 524 CB ILE 226 0.450 8.025 21.332 1.00 16.10 ATOM 525 CG2 ILE 226 -0.508 7.132 22.108 1.00 16.10 ATOM 526 CG1 ILE 226 1.729 8.293 22.132 1.00 16.10 ATOM 527 CD1 ILE 226 1.509 9.113 23.387 1.00 16.10 ATOM 528 C ILE 226 -0.499 7.094 19.213 1.00 14.61 ATOM 529 O ILE 226 -0.986 5.961 19.2001.00 16.10 ATOM 530 N THR 227 -1.042 8.123 18.569 1.00 15.93 ATOM 531 CA THR 227 -2.278 7.997 17.800 1.00 15.93 ATOM 532 CB THR 227 -2.706 9.360 17.207 1.00 22.37 ATOM 533 OG1 THR 227 -2.890 10.301 18.273 1.00 22.37 ATOM 534 CG2 THR 227 -4.0149.232 16.434 1.00 22.37 ATOM 535 C THR 227 -2.149 6.964 16.680 1.00 15.93 ATOM 536 O THR 227 -3.091 6.217 16.402 1.00 22.37 ATOM 537 N ARG 228 -0.982 6.916 16.045 1.00 14.49 ATOM 538 CA ARG 228 -0.750 5.956 14.975 1.00 14.49 ATOM 539 CB ARG 2280.602 6.188 14.307 1.00 33.87 ATOM 540 CG ARG 228 0.701 7.482 13.540 1.00 33.87 ATOM 541 CD ARG 228 2.053 7.572 12.868 1.00 33.87 ATOM 542 NE ARG 228 2.510 8.952 12.793 1.00 33.87 ATOM 543 CZ ARG 228 3.551 9.431 13.469 1.00 33.87 ATOM 544 NH1ARG 228 4.256 8.634 14.270 1.00 33.87 ATOM 545 NH2 ARG 228 3.864 10.716 13.374 1.00 33.87 ATOM 546 C ARG 228 -0.813 4.531 15.516 1.00 14.49 ATOM 547 O ARG 228 -1.309 3.632 14.839 1.00 33.87 ATOM 548 N VAL 229 -0.313 4.327 16.735 1.00 14.80 ATOM549 CA VAL 229 -0.333 3.002 17.352 1.00 14.80 ATOM 550 CB VAL 229 0.456 2.979 18.683 1.00 13.78 ATOM 551 CG1 VAL 229 0.339 1.612 19.350 1.00 13.78 ATOM 552 CG2 VAL 229 1.915 3.312 18.430 1.00 13.78 ATOM 553 C VAL 229 -1.788 2.602 17.591 1.00 14.80 ATOM 554 O VAL 229 -2.185 1.465 17.323 1.00 13.78 ATOM 555 N VAL 230 -2.588 3.561 18.047 1.00 9.33 ATOM 556 CA VAL 230 -4.005 3.327 18.292 1.00 9.33 ATOM 557 CB VAL 230 -4.679 4.564 18.909 1.00 16.07 ATOM 558 CG1 VAL 230 -6.168 4.31919.076 1.00 16.07 ATOM 559 CG2 VAL 230 -4.038 4.896 20.253 1.00 16.07 ATOM 560 C VAL 230 -4.700 2.982 16.981 1.00 9.33 ATOM 561 O VAL 230 -5.504 2.049 16.929 1.00 16.07 ATOM 562 N ASP 231 -4.364 3.719 15.922 1.00 12.71 ATOM 563 CA ASP 231-4.951 3.496 14.603 1.00 12.71 ATOM 564 CB ASP 231 -4.529 4.596 13.624 1.00 27.08 ATOM 565 CG ASP 231 -5.053 5.967 14.020 1.00 27.08 ATOM 566 OD1 ASP 231 -6.144 6.047 14.624 1.00 27.08 ATOM 567 OD2 ASP 231 -4.370 6.969 13.723 1.00 27.08 ATOM568 C ASP 231 -4.570 2.132 14.049 1.00 12.71 ATOM 569 O ASP 231 -5.413 1.436 13.483 1.00 27.08 ATOM 570 N PHE 232 -3.305 1.755 14.215 1.00 14.33 ATOM 571 CA PHE 232 -2.823 0.461 13.748 1.00 14.33 ATOM 572 CB PHE 232 -1.351 0.257 14.134 1.00 16.35 ATOM 573 CG PHE 232 -0.911 -1.184 14.097 1.00 16.35 ATOM 574 CD1 PHE 232 -0.789 -1.862 12.887 1.00 16.35 ATOM 575 CD2 PHE 232 -0.661 -1.879 15.280 1.00 16.35 ATOM 576 CE1 PHE 232 -0.430 -3.208 12.851 1.00 16.35 ATOM 577 CE2 PHE 232 -0.302-3.224 15.255 1.00 16.35 ATOM 578 CZ PHE 232 -0.187 -3.890 14.038 1.00 16.35 ATOM 579 C PHE 232 -3.670 -0.642 14.368 1.00 14.33 ATOM 580 O PHE 232 -4.226 -1.482 13.661 1.00 16.35 ATOM 581 N ALA 233 -3.769 -0.619 15.695 1.00 15.30 ATOM 582 CAALA 233 -4.537 -1.607 16.444 1.00 15.30 ATOM 583 CB ALA 233 -4.413 -1.335 17.938 1.00 12.88 ATOM 584 C ALA 233 -6.005 -1.609 16.030 1.00 15.30 ATOM 585 O ALA 233 -6.627 -2.663 15.902 1.00 12.88 ATOM 586 N LYS 234 -6.542 -0.419 15.795 1.00 25.69 ATOM 587 CA LYS 234 -7.933 -0.256 15.401 1.00 25.69 ATOM 588 CB LYS 234 -8.270 1.234 15.318 1.00 45.91 ATOM 589 CG LYS 234 -9.574 1.595 15.979 1.00 45.91 ATOM 590 CD LYS 234 -9.535 1.268 17.463 1.00 45.91 ATOM 591 CE LYS 234 -10.938 1.047 18.0061.00 45.91 ATOM 592 NZ LYS 234 -11.605 -0.106 17.327 1.00 45.91 ATOM 593 C LYS 234 -8.240 -0.931 14.067 1.00 25.69 ATOM 594 O LYS 234 -9.368 -1.368 13.827 1.00 45.91 ATOM 595 N LYS 235 -7.234 -1.019 13.204 1.00 17.44 ATOM 596 CA LYS 235 -7.406-1.627 11.892 1.00 17.44 ATOM 597 CB LYS 235 -6.459 -0.975 10.884 1.00 26.26 ATOM 598 CG LYS 235 -6.757 0.499 10.669 1.00 26.26 ATOM 599 CD LYS 235 -5.785 1.141 9.706 1.00 26.26 ATOM 600 CE LYS 235 -6.154 2.593 9.460 1.00 26.26 ATOM 601 NZ LYS235 -5.231 3.230 8.484 1.00 26.26 ATOM 602 C LYS 235 -7.258 -3.146 11.875 1.00 17.44 ATOM 603 O LYS 235 -7.365 -3.773 10.817 1.00 26.26 ATOM 604 N LEU 236 -7.015 -3.738 13.040 1.00 21.99 ATOM 605 CA LEU 236 -6.880 -5.187 13.144 1.00 21.99 ATOM606 CB LEU 236 -5.792 -5.564 14.154 1.00 25.38

ATOM 607 CG LEU 236 -4.362 -5.127 13.818 1.00 25.38 ATOM 608 CD1 LEU 236 -3.415 -5.555 14.929 1.00 25.38 ATOM 609 CD2 LEU 236 -3.931 -5.725 12.491 1.00 25.38 ATOM 610 C LEU 236 -8.219 -5.796 13.556 1.00 21.99 ATOM 611 O LEU 236 -8.821-5.386 14.553 1.00 25.38 ATOM 612 N PRO 237 -8.682 -6.819 12.817 1.00 34.89 ATOM 613 CD PRO 237 -7.936 -7.474 11.730 1.00 42.99 ATOM 614 CA PRO 237 -9.953 -7.513 13.071 1.00 34.89 ATOM 615 CB PRO 237 -9.911 -8.687 12.084 1.00 42.99 ATOM 616 CGPRO 237 -8.433 -8.887 11.816 1.00 42.99 ATOM 617 C PRO 237 -10.184 -7.986 14.513 1.00 34.89 ATOM 618 O PRO 237 -11.142 -7.563 15.159 1.00 42.99 ATOM 619 N MET 238 -9.301 -8.843 15.021 1.00 40.45 ATOM 620 CA MET 238 -9.433 -9.364 16.382 1.00 40.45 ATOM 621 CB MET 238 -8.360 -10.423 16.671 1.00 59.70 ATOM 622 CG MET 238 -8.689 -11.839 16.195 1.00 59.70 ATOM 623 SD MET 238 -8.013 -12.275 14.573 1.00 59.70 ATOM 624 CE MET 238 -6.482 -13.074 15.032 1.00 59.70 ATOM 625 C MET 238 -9.395-8.305 17.486 1.00 40.45 ATOM 626 O MET 238 -9.801 -8.574 18.617 1.00 59.70 ATOM 627 N PHE 239 -8.928 -7.103 17.160 1.00 33.70 ATOM 628 CA PHE 239 -8.829 -6.037 18.152 1.00 33.70 ATOM 629 CB PHE 239 -7.651 -5.113 17.829 1.00 22.27 ATOM 630 CGPHE 239 -7.386 -4.079 18.885 1.00 22.27 ATOM 631 CD1 PHE 239 -6.602 -4.385 19.990 1.00 22.27 ATOM 632 CD2 PHE 239 -7.926 -2.802 18.778 1.00 22.27 ATOM 633 CE1 PHE 239 -6.358 -3.436 20.974 1.00 22.27 ATOM 634 CE2 PHE 239 -7.688 -1.846 19.757 1.00 22.27 ATOM 635 CZ PHE 239 -6.901 -2.163 20.857 1.00 22.27 ATOM 636 C PHE 239 -10.103 -5.213 18.329 1.00 33.70 ATOM 637 O PHE 239 -10.594 -5.059 19.446 1.00 22.27 ATOM 638 N SER 240 -10.629 -4.679 17.232 1.00 23.42 ATOM 639 CA SER 240 -11.837-3.857 17.278 1.00 23.42 ATOM 640 CB SER 240 -12.175 -3.352 15.884 1.00 26.21 ATOM 641 C SER 240 -13.046 -4.562 17.899 1.00 23.42 ATOM 642 O SER 240 -13.976 -3.909 18.369 1.00 26.21 ATOM 643 N GLU 241 -13.028 -5.891 17.893 1.00 26.54 ATOM 644CA GLU 241 -14.116 -6.695 18.450 1.00 26.54 ATOM 645 CB GLU 241 -14.007 -8.139 17.957 1.00 67.32 ATOM 646 CG GLU 241 -14.241 -8.322 16.467 1.00 67.32 ATOM 647 CD GLU 241 -13.979 -9.748 16.001 1.00 67.32 ATOM 648 OE1 GLU 241 -14.161 -10.691 16.8031.00 67.32 ATOM 649 OE2 GLU 241 -13.584 -9.924 14.828 1.00 67.32 ATOM 650 C GLU 241 -14.137 -6.706 19.975 1.00 26.54 ATOM 651 O GLU 241 -15.182 -6.924 20.589 1.00 67.32 ATOM 652 N LEU 242 -12.972 -6.506 20.579 1.00 26.16 ATOM 653 CA LEU 242-12.835 -6.514 22.030 1.00 26.16 ATOM 654 CB LEU 242 -11.352 -6.473 22.412 1.00 19.79 ATOM 655 CG LEU 242 -10.461 -7.627 21.956 1.00 19.79 ATOM 656 CD1 LEU 242 -9.014 -7.309 22.264 1.00 19.79 ATOM 657 CD2 LEU 242 -10.888 -8.912 22.640 1.00 19.79 ATOM 658 C LEU 242 -13.547 -5.351 22.711 1.00 26.16 ATOM 659 O LEU 242 -13.738 -4.290 22.115 1.00 19.79 ATOM 660 N PRO 243 -13.980 -5.547 23.968 1.00 17.98 ATOM 661 CD PRO 243 -13.996 -6.785 24.764 1.00 19.17 ATOM 662 CA PRO 243 -14.657 -4.45424.671 1.00 17.98 ATOM 663 CB PRO 243 -15.095 -5.105 25.988 1.00 19.17 ATOM 664 CG PRO 243 -14.155 -6.263 26.161 1.00 19.17 ATOM 665 C PRO 243 -13.652 -3.323 24.898 1.00 17.98 ATOM 666 O PRO 243 -12.458 -3.572 25.081 1.00 19.17 ATOM 667 N CYS244 -14.142 -2.088 24.880 1.00 20.08 ATOM 668 CA CYS 244 -13.310 -0.900 25.059 1.00 20.08 ATOM 669 CB CYS 244 -14.194 0.329 25.278 1.00 61.80 ATOM 670 SG CYS 244 -13.674 1.784 24.340 1.00 61.80 ATOM 671 C CYS 244 -12.286 -1.017 26.189 1.00 20.08 ATOM 672 O CYS 244 -11.141 -0.590 26.040 1.00 61.80 ATOM 673 N GLU 245 -12.691 -1.630 27.299 1.00 21.05 ATOM 674 CA GLU 245 -11.814 -1.811 28.454 1.00 21.05 ATOM 675 CB GLU 245 -12.541 -2.560 29.578 1.00 40.41 ATOM 676 CG GLU 245 -13.510 -1.70530.393 1.00 40.41 ATOM 677 CD GLU 245 -14.953 -1.773 29.910 1.00 40.41 ATOM 678 OE1 GLU 245 -15.854 -1.761 30.775 1.00 40.41 ATOM 679 OE2 GLU 245 -15.197 -1.824 28.683 1.00 40.41 ATOM 680 C GLU 245 -10.541 -2.558 28.084 1.00 21.05 ATOM 681 OGLU 245 -9.439 -2.138 28.440 1.00 40.41 ATOM 682 N ASP 246 -10.698 -3.654 27.351 1.00 17.22 ATOM 683 CA ASP 246 -9.564 -4.463 26.924 1.00 17.22 ATOM 684 CB ASP 246 -10.044 -5.774 26.303 1.00 30.41 ATOM 685 CG ASP 246 -10.634 -6.727 27.327 1.00 30.41 ATOM 686 OD1 ASP 246 -10.755 -6.349 28.512 1.00 30.41 ATOM 687 OD2 ASP 246 -10.975 -7.864 26.946 1.00 30.41 ATOM 688 C ASP 246 -8.693 -3.705 25.936 1.00 17.22 ATOM 689 O ASP 246 -7.467 -3.713 26.050 1.00 30.41 ATOM 690 N GLN 247 -9.332-3.045 24.973 1.00 17.12 ATOM 691 CA GLN 247 -8.615 -2.272 23.966 1.00 17.12 ATOM 692 CB GLN 247 -9.594 -1.494 23.088 1.00 16.72 ATOM 693 CG GLN 247 -10.504 -2.365 22.242 1.00 16.72 ATOM 694 CD GLN 247 -11.352 -1.553 21.290 1.00 16.72 ATOM 695OE1 GLN 247 -10.925 -0.515 20.790 1.00 16.72 ATOM 696 NE2 GLN 247 -12.560 -2.018 21.033 1.00 16.72 ATOM 697 C GLN 247 -7.650 -1.303 24.637 1.00 17.12 ATOM 698 O GLN 247 -6.476 -1.228 24.273 1.00 16.72 ATOM 699 N ILE 248 -8.152 -0.591 25.640 1.00 19.19 ATOM 700 CA ILE 248 -7.358 0.377 26.387 1.00 19.19 ATOM 701 CB ILE 248 -8.238 1.137 27.410 1.00 24.32 ATOM 702 CG2 ILE 248 -7.385 2.055 28.282 1.00 24.32 ATOM 703 CG1 ILE 248 -9.312 1.942 26.668 1.00 24.32 ATOM 704 CD1 ILE 248 -10.3272.618 27.573 1.00 24.32 ATOM 705 C ILE 248 -6.180 -0.297 27.093 1.00 19.19 ATOM 706 O ILE 248 -5.035 0.131 26.943 1.00 24.32 ATOM 707 N ILE 249 -6.457 -1.367 27.830 1.00 12.09 ATOM 708 CA ILE 249 -5.409 -2.090 28.547 1.00 12.09 ATOM 709 CB ILE249 -5.996 -3.295 29.322 1.00 30.01 ATOM 710 CG2 ILE 249 -4.884 -4.168 29.885 1.00 30.01 ATOM 711 CG1 ILE 249 -6.899 -2.794 30.451 1.00 30.01 ATOM 712 CD1 ILE 249 -7.598 -3.893 31.215 1.00 30.01 ATOM 713 C ILE 249 -4.299 -2.561 27.602 1.00 12.09 ATOM 714 O ILE 249 -3.115 -2.339 27.866 1.00 30.01 ATOM 715 N LEU 250 -4.691 -3.168 26.486 1.00 20.87 ATOM 716 CA LEU 250 -3.740 -3.669 25.498 1.00 20.87 ATOM 717 CB LEU 250 -4.474 4.410 24.376 1.00 15.15 ATOM 718 CG LEU 250 -5.252 -5.669 24.7611.00 15.15 ATOM 719 CD1 LEU 250 -5.907 -6.256 23.533 1.00 15.15 ATOM 720 CD2 LEU 250 -4.325 -6.686 25.400 1.00 15.15 ATOM 721 C LEU 250 -2.900 -2.548 24.902 1.00 20.87 ATOM 722 O LEU 250 -1.680 -2.667 24.792 1.00 15.15 ATOM 723 N LEU 251 -3.559-1.455 24.532 1.00 9.31 ATOM 724 CA LEU 251 -2.887 -0.301 23.945 1.00 9.31 ATOM 725 CB LEU 251 -3.920 0.760 23.553 1.00 19.90 ATOM 726 CG LEU 251 -4.075 1.127 22.073 1.00 19.90 ATOM 727 CD1 LEU 251 -3.281 0.190 21.180 1.00 19.90 ATOM 728 CD2LEU 251 -5.550 1.113 21.699 1.00 19.90 ATOM 729 C LEU 251 -1.851 0.307 24.887 1.00 9.31 ATOM 730 O LEU 251 -0.699 0.521 24.507 1.00 19.90 ATOM 731 N LYS 252 -2.253 0.545 26.127 1.00 18.83 ATOM 732 CA LYS 252 -1.362 1.132 27.114 1.00

18.83 ATOM 733 CB LYS 252 -2.138 1.455 28.395 1.00 42.69 ATOM 734 CG LYS 252 -3.395 2.274 28.130 1.00 42.69 ATOM 735 CD LYS 252 -3.588 3.412 29.115 1.00 42.69 ATOM 736 CE LYS 252 -3.998 2.934 30.493 1.00 42.69 ATOM 737 NZ LYS 252 4.3004.109 31.361 1.00 42.69 ATOM 738 C LYS 252 -0.171 0.222 27.408 1.00 18.83 ATOM 739 O LYS 252 0.942 0.700 27.646 1.00 42.69 ATOM 740 N GLY 253 -0.392 -1.086 27.328 1.00 16.16 ATOM 741 CA GLY 253 0.676 -2.031 27.595 1.00 16.16 ATOM 742 C GLY 2531.688 -2.232 26.479 1.00 16.16 ATOM 743 O GLY 253 2.836 -2.587 26.747 1.00 34.57 ATOM 744 N CYS 254 1.286 -1.999 25.233 1.00 21.81 ATOM 745 CA CYS 254 2.194 -2.203 24.108 1.00 21.81 ATOM 746 CB CYS 254 1.563 -3.151 23.093 1.00 23.60 ATOM 747 SGCYS 254 0.211 -2.387 22.179 1.00 23.60 ATOM 748 C CYS 254 2.616 -0.935 23.380 1.00 21.81 ATOM 749 O CYS 254 3.499 -0.983 22.521 1.00 23.60 ATOM 750 N CYS 255 2.004 0.193 23.724 1.00 14.98 ATOM 751 CA CYS 255 2.309 1.461 23.066 1.00 14.98 ATOM752 CB CYS 255 1.611 2.616 23.781 1.00 24.32 ATOM 753 SG CYS 255 1.602 4.153 22.841 1.00 24.32 ATOM 754 C CYS 255 3.804 1.750 22.922 1.00 14.98 ATOM 755 O CYS 255 4.305 1.895 21.805 1.00 24.32 ATOM 756 N MET 256 4.525 1.777 24.037 1.00 13.77 ATOM 757 CA MET 256 5.959 2.056 24.003 1.00 13.77 ATOM 758 CB MET 256 6.515 2.218 25.423 1.00 19.23 ATOM 759 CG MET 256 7.988 2.607 25.477 1.00 19.23 ATOM 760 SD MET 256 8.344 4.132 24.571 1.00 19.23 ATOM 761 CE MET 256 10.127 4.254 24.782 1.00 19.23 ATOM 762 C MET 256 6.734 0.978 23.246 1.00 13.77 ATOM 763 O MET 256 7.672 1.284 22.516 1.00 19.23 ATOM 764 N GLU 257 6.316 -0.275 23.400 1.00 12.57 ATOM 765 CA GLU 257 6.971 -1.397 22.730 1.00 12.57 ATOM 766 CB GLU 257 6.342 -2.716 23.1821.00 31.54 ATOM 767 CG GLU 257 6.497 -2.982 24.677 1.00 31.54 ATOM 768 CD GLU 257 5.720 -4.196 25.167 1.00 31.54 ATOM 769 OE1 GLU 257 5.220 -4.983 24.334 1.00 31.54 ATOM 770 OE2 GLU 257 5.607 -4.361 26.400 1.00 31.54 ATOM 771 C GLU 257 6.889-1.254 21.211 1.00 12.57 ATOM 772 O GLU 257 7.881 -1.452 20.505 1.00 31.54 ATOM 773 N ILE 258 5.712 -0.881 20.717 1.00 17.89 ATOM 774 CA ILE 258 5.508 -0.692 19.288 1.00 17.89 ATOM 775 CB ILE 258 4.001 -0.555 18.946 1.00 15.57 ATOM 776 CG2 ILE258 3.813 -0.129 17.493 1.00 15.57 ATOM 777 CG1 ILE 258 3.288 -1.886 19.211 1.00 15.57 ATOM 778 CD1 ILE 258 1.798 -1.872 18.922 1.00 15.57 ATOM 779 C ILE 258 6.289 0.535 18.811 1.00 17.89 ATOM 780 O ILE 258 7.000 0.468 17.805 1.00 15.57 ATOM781 N MET 259 6.196 1.636 19.556 1.00 11.23 ATOM 782 CA MET 259 6.907 2.861 19.201 1.00 11.23 ATOM 783 CB MET 259 6.568 3.995 20.175 1.00 22.19 ATOM 784 CG MET 259 5.112 4.439 20.117 1.00 22.19 ATOM 785 SD MET 259 4.828 6.033 20.915 1.00 22.19 ATOM 786 CE MET 259 5.038 5.606 22.621 1.00 22.19 ATOM 787 C MET 259 8.415 2.637 19.131 1.00 11.23 ATOM 788 O MET 259 9.060 3.008 18.145 1.00 22.19 ATOM 789 N SER 260 8.974 1.994 20.153 1.00 8.59 ATOM 790 CA SER 260 10.408 1.706 20.195 1.00 8.59 ATOM 791 CB SER 260 10.763 0.939 21.472 1.00 23.39 ATOM 792 OG SER 260 10.430 1.685 22.623 1.00 23.39 ATOM 793 C SER 260 10.793 0.864 18.977 1.00 8.59 ATOM 794 O SER 260 11.824 1.100 18.350 1.00 23.39 ATOM 795 N LEU 261 9.952 -0.111 18.644 1.00 13.26 ATOM 796 CA LEU 261 10.194 -0.992 17.507 1.00 13.26 ATOM 797 CB LEU 261 9.076 -2.035 17.401 1.00 14.32 ATOM 798 CG LEU 261 9.019 -2.894 16.134 1.00 14.32 ATOM 799 CD1 LEU 261 10.278 -3.733 15.999 1.00 14.32 ATOM 800 CD2 LEU 261 7.785-3.772 16.174 1.00 14.32 ATOM 801 C LEU 261 10.276 -0.170 16.220 1.00 13.26 ATOM 802 O LEU 261 11.213 -0.313 15.432 1.00 14.32 ATOM 803 N ARG 262 9.330 0.744 16.043 1.00 10.57 ATOM 804 CA ARG 262 9.278 1.598 14.861 1.00 10.57 ATOM 805 CB ARG262 8.018 2.454 14.917 1.00 16.08 ATOM 806 CG ARG 262 6.755 1.647 14.728 1.00 16.08 ATOM 807 CD ARG 262 5.540 2.525 14.614 1.00 16.08 ATOM 808 NE ARG 262 4.418 1.765 14.076 1.00 16.08 ATOM 809 CZ ARG 262 3.260 2.289 13.689 1.00 16.08 ATOM 810NH1 ARG 262 3.050 3.596 13.780 1.00 16.08 ATOM 811 NH2 ARG 262 2.322 1.497 13.183 1.00 16.08 ATOM 812 C ARG 262 10.530 2.471 14.704 1.00 10.57 ATOM 813 O ARG 262 11.038 2.649 13.589 1.00 16.08 ATOM 814 N ALA 263 11.016 3.014 15.820 1.00 13.37 ATOM 815 CA ALA 263 12.221 3.842 15.831 1.00 13.37 ATOM 816 CB ALA 263 12.363 4.516 17.172 1.00 17.12 ATOM 817 C ALA 263 13.443 2.964 15.561 1.00 13.37 ATOM 818 O ALA 263 14.313 3.316 14.762 1.00 17.12 ATOM 819 N ALA 264 13.474 1.802 16.207 1.00 16.55 ATOM 820 CA ALA 264 14.514 0.855 16.072 1.00 16.55 ATOM 821 CB ALA 264 14.375 -0.327 17.019 1.00 24.62 ATOM 822 C ALA 264 14.770 0.364 14.642 1.00 16.55 ATOM 823 O ALA 264 15.904 0.244 14.169 1.00 24.62 ATOM 824 N VAL 265 13.670 0.07313.955 1.00 22.25 ATOM 825 CA VAL 265 13.754 -0.401 12.583 1.00 22.25 ATOM 826 CB VAL 265 12.428 -1.038 12.086 1.00 25.31 ATOM 827 CG1 VAL 265 12.079 -2.239 12.936 1.00 25.31 ATOM 828 CG2 VAL 265 11.302 -0.030 12.091 1.00 25.31 ATOM 829 C VAL265 14.208 0.707 11.639 1.00 22.25 ATOM 830 O VAL 265 14.615 0.434 10.513 1.00 25.31 ATOM 831 N ARG 266 14.124 1.955 12.092 1.00 26.45 ATOM 832 CA ARG 266 14.567 3.086 11.283 1.00 26.45 ATOM 833 CB ARG 266 13.596 4.261 11.399 1.00 38.04 ATOM834 CG ARG 266 12.232 4.019 10.807 1.00 38.04 ATOM 835 CD ARG 266 11.503 5.339 10.651 1.00 38.04 ATOM 836 NE ARG 266 10.074 5.216 10.925 1.00 38.04 ATOM 837 CZ ARG 266 9.504 5.551 12.079 1.00 38.04 ATOM 838 NH1 ARG 266 10.237 6.038 13.075 1.00 38.04 ATOM 839 NH2 ARG 266 8.196 5.411 12.240 1.00 38.04 ATOM 840 C ARG 266 15.957 3.531 11.729 1.00 26.45 ATOM 841 O ARG 266 16.296 4.717 11.660 1.00 38.04 ATOM 842 N TYR 267 16.733 2.590 12.251 1.00 24.87 ATOM 843 CA TYR 267 18.083 2.88812.700 1.00 24.87 ATOM 844 CB TYR 267 18.592 1.788 13.639 1.00 25.84 ATOM 845 CG TYR 267 20.073 1.865 13.931 1.00 25.84 ATOM 846 CD1 TYR 267 20.579 2.789 14.844 1.00 25.84 ATOM 847 CE1 TYR 267 21.940 2.865 15.103 1.00 25.84 ATOM 848 CD2 TYR 26720.971 1.017 13.284 1.00 25.84 ATOM 849 CE2 TYR 267 22.331 1.085 13.536 1.00 25.84 ATOM 850 CZ TYR 267 22.810 2.011 14.444 1.00 25.84 ATOM 851 OH TYR 267 24.162 2.078 14.683 1.00 25.84 ATOM 852 C TYR 267 18.999 3.009 11.488 1.00 24.87 ATOM 853O TYR 267 19.019 2.130 10.625 1.00 25.84 ATOM 854 N ASP 268 19.751 4.102 11.423 1.00 28.13 ATOM 855 CA ASP 268 20.666 4.320 10.313 1.00 28.13 ATOM 856 CB ASP 268 20.524 5.744 9.773 1.00 51.63 ATOM 857 CG ASP 268 21.339 5.973 8.517 1.00 51.63

ATOM 858 OD1 ASP 268 21.060 5.305 7.498 1.00 51.63 ATOM 859 OD2 ASP 268 22.262 6.814 8.547 1.00 51.63 ATOM 860 C ASP 268 22.105 4.068 10.749 1.00 28.13 ATOM 861 O ASP 268 22.683 4.854 11.500 1.00 51.63 ATOM 862 N PRO 269 22.707 2.96410.276 1.00 37.07 ATOM 863 CD PRO 269 22.103 1.938 9.410 1.00 39.18 ATOM 864 CA PRO 269 24.086 2.612 10.623 1.00 37.07 ATOM 865 CB PRO 269 24.319 1.324 9.832 1.00 39.18 ATOM 866 CG PRO 269 22.950 0.735 9.706 1.00 39.18 ATOM 867 C PRO 269 25.0793.698 10.216 1.00 37.07 ATOM 868 O PRO 269 26.003 4.006 10.964 1.00 39.18 ATOM 869 N ALA 270 24.855 4.295 9.047 1.00 46.88 ATOM 870 CA ALA 279 25.730 5.340 8.519 1.00 46.88 ATOM 871 CB ALA 270 25.177 5.873 7.198 1.00 41.71 ATOM 872 C ALA 27025.974 6.493 9.492 1.00 46.88 ATOM 873 O ALA 270 27.121 6.844 9.763 1.00 41.71 ATOM 874 N SER 271 24.899 7.081 10.009 1.00 34.54 ATOM 875 CA SER 271 25.013 8.198 10.941 1.00 34.54 ATOM 876 CB SER 271 23.959 9.259 10.618 1.00 42.29 ATOM 877 OGSER 271 22.686 8.668 10.422 1.00 42.29 ATOM 878 C SER 271 24.910 7.793 12.408 1.00 34.54 ATOM 879 O SER 271 25.169 8.607 13.297 1.00 42.29 ATOM 880 N ASP 272 24.546 6.535 12.653 1.00 41.05 ATOM 881 CA ASP 272 24.388 6.005 14.007 1.00 41.05 ATOM882 CB ASP 272 25.720 6.078 14.772 1.00 47.32 ATOM 883 CG ASP 272 25.653 5.428 16.147 1.00 47.32 ATOM 884 OD1 ASP 272 24.981 4.384 16.299 1.00 47.32 ATOM 885 OD2 ASP 272 26.284 5.967 17.081 1.00 47.32 ATOM 886 C ASP 272 23.279 6.777 14.730 1.00 41.05 ATOM 887 O ASP 272 23.444 7.233 15.866 1.00 47.32 ATOM 888 N THR 273 22.139 6.905 14.058 1.00 27.60 ATOM 889 CA THR 273 20.996 7.618 14.608 1.00 27.60 ATOM 890 CB THR 273 20.808 8.991 13.911 1.00 30.96 ATOM 891 OG1 THR 273 20.723 8.80812.491 1.00 30.96 ATOM 892 CG2 THR 273 21.967 9.924 14.228 1.00 30.96 ATOM 893 C THR 273 19.701 6.829 14.442 1.00 27.60 ATOM 894 O THR 273 19.633 5.883 13.650 1.00 30.96 ATOM 895 N LEU 274 18.696 7.192 15.232 1.00 20.89 ATOM 896 CA LEU 27417.374 6.574 15.161 1.00 20.89 ATOM 897 CB LEU 274 16.862 6.193 16.555 1.00 22.48 ATOM 898 CG LEU 274 17.480 5.009 17.301 1.00 22.48 ATOM 899 CD1 LEU 274 16.798 4.866 18.650 1.00 22.48 ATOM 900 CD2 LEU 274 17.317 3.736 16.497 1.00 22.48 ATOM901 C LEU 274 16.470 7.654 14.586 1.00 20.89 ATOM 902 O LEU 274 16.753 8.842 14.744 1.00 22.48 ATOM 903 N THR 275 15.393 7.258 13.922 1.00 27.89 ATOM 904 CA THR 275 14.478 8.235 13.354 1.00 27.89 ATOM 905 CB THR 275 14.325 8.045 11.832 1.00 37.64 ATOM 906 OG1 THR 275 15.622 7.983 11.228 1.00 37.64 ATOM 907 CG2 THR 275 13.570 9.215 11.222 1.00 37.64 ATOM 908 C THR 275 13.120 8.135 14.032 1.00 27.89 ATOM 909 O THR 275 12.493 7.081 14.019 1.00 37.64 ATOM 910 N LEU 276 12.700 9.22614.667 1.00 28.07 ATOM 911 CA LEU 276 11.418 9.275 15.358 1.00 28.07 ATOM 912 CB LEU 276 11.497 10.214 16.572 1.00 24.81 ATOM 913 CG LEU 276 12.639 10.005 17.577 1.00 24.81 ATOM 914 CD1 LEU 276 12.459 10.929 18.769 1.00 24.81 ATOM 915 CD2 LEU276 12.692 8.558 18.038 1.00 24.81 ATOM 916 C LEU 276 10.339 9.761 14.395 1.00 28.07 ATOM 917 O LEU 276 10.533 10.760 13.691 1.00 24.81 ATOM 918 N SER 277 9.232 9.027 14.331 1.00 29.24 ATOM 919 CA SER 277 8.106 9.357 13.458 1.00 29.24 ATOM 920CB SER 277 7.369 10.594 13.985 1.00 30.56 ATOM 921 OG SER 277 6.845 10.358 15.283 1.00 30.56 ATOM 922 C SER 277 8.533 9.569 12.005 1.00 29.24 ATOM 923 O SER 277 7.902 10.326 11.263 1.00 30.56 ATOM 924 N GLY 278 9.619 8.908 11.618 1.00 34.41 ATOM 925 CA GLY 278 10.135 9.024 10.263 1.00 34.41 ATOM 926 C GLY 278 10.472 10.442 9.830 1.00 34.41 ATOM 927 O GLY 278 10.516 10.725 8.631 1.00 44.04 ATOM 928 N GLU 279 10.733 11.326 10.791 1.00 37.82 ATOM 929 CA GLU 279 11.056 12.717 10.4791.00 37.82 ATOM 930 CB GLU 279 9.808 13.600 10.612 1.00 70.24 ATOM 931 CG GLU 279 9.202 13.631 12.014 1.00 70.24 ATOM 932 CD GLU 279 8.028 14.593 12.141 1.00 70.24 ATOM 933 OE1 GLU 279 8.028 15.406 13.093 1.00 70.24 ATOM 934 OE2 GLU 279 7.10314.535 11.301 1.00 70.24 ATOM 935 C GLU 279 12.192 13.321 11.300 1.00 37.82 ATOM 936 O GLU 279 12.857 14.248 10.841 1.00 70.24 ATOM 937 N MET 280 12.424 12.811 12.505 1.00 33.77 ATOM 938 CA MET 280 13.482 13.360 13.344 1.00 33.77 ATOM 939 CBMET 280 12.903 13.848 14.674 1.00 33.89 ATOM 940 CG MET 280 13.898 14.595 15.545 1.00 33.89 ATOM 941 SD MET 280 13.350 14.740 17.256 1.00 33.89 ATOM 942 CE MET 280 12.100 16.017 17.121 1.00 33.89 ATOM 943 C MET 280 14.620 12.383 13.613 1.00 33.77 ATOM 944 O MET 280 14.432 11.366 14.282 1.00 33.89 ATOM 945 N ALA 281 15.797 12.690 13.080 1.00 30.24 ATOM 946 CA ALA 281 16.972 11.852 13.287 1.00 30.24 ATOM 947 CB ALA 281 17.937 11.998 12.120 1.00 25.10 ATOM 948 C ALA 281 17.631 12.30914.587 1.00 30.24 ATOM 949 O ALA 281 18.008 13.477 14.718 1.00 25.10 ATOM 950 N VAL 282 17.743 11.401 15.551 1.00 32.12 ATOM 951 CA VAL 282 18.339 11.726 16.844 1.00 32.12 ATOM 952 CB VAL 282 17.303 11.606 17.991 1.00 37.75 ATOM 953 CG1 VAL 28216.184 12.615 17.799 1.00 37.75 ATOM 954 CG2 VAL 282 16.739 10.193 18.055 1.00 37.75 ATOM 955 C VAL 282 19.543 10.852 17.181 1.00 32.12 ATOM 956 O VAL 282 19.614 9.690 16.778 1.00 37.75 ATOM 957 N LYS 283 20.491 11.428 17.913 1.00 26.82 ATOM958 CA LYS 283 21.700 10.722 18.328 1.00 26.82 ATOM 959 CB LYS 283 22.894 11.679 18.342 1.00 57.25 ATOM 960 CG LYS 283 23.258 12.245 16.979 1.00 57.25 ATOM 961 CD LYS 283 24.282 13.361 17.105 1.00 57.25 ATOM 962 CE LYS 283 24.752 13.836 15.7411.00 57.25 ATOM 963 NZ LYS 283 25.518 12.772 15.033 1.00 57.25 ATOM 964 C LYS 283 21.509 10.120 19.717 1.00 26.82 ATOM 965 O LYS 283 20.648 10.566 20.477 1.00 57.25 ATOM 966 N ARG 284 22.351 9.146 20.058 1.00 26.41 ATOM 967 CA ARG 284 22.2978.457 21.351 1.00 26.41 ATOM 968 CB ARG 284 23.527 7.566 21.528 1.00 41.02 ATOM 969 CG ARG 284 23.715 6.539 20.440 1.00 41.02 ATOM 970 CD ARG 284 25.016 5.794 20.616 1.00 41.02 ATOM 971 NE ARG 284 25.145 4.730 19.630 1.00 41.02 ATOM 972 CZ ARG284 24.759 3.475 19.831 1.00 41.02 ATOM 973 NH1 ARG 284 24.221 3.117 20.990 1.00 41.02 ATOM 974 NH2 ARG 284 24.886 2.584 18.859 1.00 41.02 ATOM 975 C ARG 284 22.200 9.399 22.543 1.00 26.41 ATOM 976 O ARG 284 21.296 9.278 23.370 1.00 41.02 ATOM977 N GLU 285 23.152 10.321 22.634 1.00 33.23 ATOM 978 CA GLU 285 23.201 11.292 23.721 1.00 33.23 ATOM 979 CB GLU 285 24.366 12.258 23.492 1.00 69.82 ATOM 980 CG GLU 285 24.485 13.359 24.533 1.00 69.82 ATOM 981 CD GLU 285 25.079 14.636 23.9641.00 69.82 ATOM 982 OE1 GLU 285 26.309 14.826 24.070 1.00 69.82 ATOM 983 OE2 GLU 285 24.309 15.453 23.409 1.00

69.82 ATOM 984 C GLU 285 21.898 12.082 23.823 1.00 33.23 ATOM 985 O GLU 285 21.336 12.239 24.907 1.00 69.82 ATOM 986 N GLN 286 21.414 12.551 22.677 1.00 28.07 ATOM 987 CA GLN 286 20.194 13.346 22.614 1.00 28.07 ATOM 988 CB GLN 28619.948 13.824 21.181 1.00 41.05 ATOM 989 CG GLN 286 21.051 14.726 20.639 1.00 41.05 ATOM 990 CD GLN 286 20.808 15.154 19.202 1.00 41.05 ATOM 991 OE1 GLN 286 20.783 14.322 18.293 1.00 41.05 ATOM 992 NE2 GLN 286 20.635 16.452 18.990 1.00 41.05 ATOM 993 C GLN 286 18.955 12.642 23.162 1.00 28.07 ATOM 994 O GLN 286 18.281 13.174 24.048 1.00 41.05 ATOM 995 N LEU 287 18.663 11.447 22.658 1.00 30.11 ATOM 996 CN LEU 287 17.492 10.705 23.116 1.00 30.11 ATOM 997 CB LEU 287 17.232 9.489 22.2191.00 21.70 ATOM 998 CG LEU 287 15.859 8.821 22.357 1.00 21.70 ATOM 999 CD1 LEU 287 14.748 9.818 22.061 1.00 21.70 ATOM 1000 CD2 LEU 287 15.763 7.628 21.421 1.00 21.70 ATOM 1001 C LEU 287 17.641 10.277 24.577 1.00 30.11 ATOM 1002 O LEU 28716.655 10.212 25.320 1.00 21.70 ATOM 1003 N LYS 288 18.878 10.015 24.992 1.00 20.72 ATOM 1004 CA LYS 288 19.156 9.611 26.365 1.00 20.72 ATOM 1005 CB LYS 288 20.626 9.213 26.514 1.00 43.14 ATOM 1006 CG LYS 288 20.991 8.721 27.903 1.00 43.14 ATOM1007 CD LYS 288 22.374 8.102 27.931 1.00 43.14 ATOM 1008 CE LYS 288 22.615 7.379 29.250 1.00 43.14 ATOM 1009 NZ LYS 288 23.866 6.568 29.224 1.00 43.14 ATOM 1010 C LYS 288 18.819 10.742 27.331 1.00 20.72 ATOM 1011 O LYS 288 18.027 10.566 28.2611.00 43.14 ATOM 1012 N ASN 289 19.380 11.917 27.067 1.00 33.64 ATOM 1013 CA ASN 289 19.156 13.090 27.906 1.00 33.64 ATOM 1014 CB ASN 289 20.190 14.173 27.590 1.00 35.61 ATOM 1015 CG ASN 289 21.607 13.730 27.898 1.00 35.61 ATOM 1016 OD1 ASN 28921.835 12.920 28.797 1.00 35.61 ATOM 1017 ND2 ASN 289 22.566 14.253 27.149 1.00 35.61 ATOM 1018 C ASN 289 17.747 13.654 27.757 1.00 33.64 ATOM 1019 O ASN 289 17.276 14.399 28.616 1.00 35.61 ATOM 1020 N GLY 290 17.072 13.287 26.672 1.00 22.05 ATOM 1021 CA GLY 290 15.722 13.767 26.435 1.00 22.05 ATOM 1022 C GLY 290 14.688 13.247 27.416 1.00 22.05 ATOM 1023 O GLY 290 13.550 13.710 27.420 1.00 29.95 ATOM 1024 N GLY 291 15.072 12.276 28.239 1.00 24.91 ATOM 1025 CA GLY 291 14.142 11.73229.211 1.00 24.91 ATOM 1026 C GLY 291 14.093 10.217 29.248 1.00 24.91 ATOM 1027 O GLY 291 13.536 9.640 30.179 1.00 29.39 ATOM 1028 N LEU 292 14.676 9.567 28.246 1.00 30.21 ATOM 1029 CA LEU 292 14.675 8.110 28.189 1.00 30.21 ATOM 1030 CB LEU 29214.732 7.626 26.734 1.00 21.45 ATOM 1031 CG LEU 292 13.439 7.795 25.928 1.00 21.45 ATOM 1032 CD1 LEU 292 13.612 7.225 24.542 1.00 21.45 ATOM 1033 CD2 LEU 292 12.296 7.087 26.630 1.00 21.45 ATOM 1034 C LEU 292 15.785 7.461 29.013 1.00 30.21 ATOM1035 O LEU 292 15.645 6.324 29.473 1.00 21.45 ATOM 1036 N GLY 293 16.885 8.180 29.205 1.00 16.29 ATOM 1037 CA GLY 293 17.992 7.638 29.970 1.00 16.29 ATOM 1038 C GLY 293 18.534 6.374 29.332 1.00 16.29 ATOM 1039 O GLY 293 18.763 6.334 28.122 1.00 25.88 ATOM 1040 N VAL 294 18.689 5.322 30.130 1.00 33.05 ATOM 1041 CA VAL 294 19.211 4.050 29.635 1.00 33.05 ATOM 1042 CB VAL 294 19.530 3.069 30.788 1.00 30.11 ATOM 1043 CG1 VAL 294 20.718 3.577 31.582 1.00 30.11 ATOM 1044 CG2 VAL 294 18.3152.887 31.697 1.00 30.11 ATOM 1045 C VAL 294 18.302 3.361 28.617 1.00 33.05 ATOM 1046 O VAL 294 18.768 2.545 27.817 1.00 30.11 ATOM 1047 N VAL 295 17.014 3.699 28.635 1.00 18.14 ATOM 1048 CA VAL 295 16.056 3.118 27.698 1.00 18.14 ATOM 1049 CBVAL 295 14.638 3.698 27.902 1.00 28.34 ATOM 1050 CG1 VAL 295 13.668 3.099 26.893 1.00 28.34 ATOM 1051 CG2 VAL 295 14.159 3.431 29.317 1.00 28.34 ATOM 1052 C VAL 295 16.521 3.415 26.275 1.00 18.14 ATOM 1053 O VAL 295 16.395 2.577 25.383 1.00 28.34 ATOM 1054 N SER 296 17.091 4.601 26.085 1.00 20.84 ATOM 1055 CA SER 296 17.596 5.028 24.785 1.00 20.84 ATOM 1056 CB SER 296 18.160 6.446 24.884 1.00 25.61 ATOM 1057 OG SER 296 18.615 6.911 23.627 1.00 25.61 ATOM 1058 C SER 296 18.687 4.07424.307 1.00 20.84 ATOM 1059 O SER 296 18.723 3.691 23.133 1.00 25.61 ATOM 1060 N ASP 297 19.571 3.691 25.224 1.00 28.08 ATOM 1061 CA ASP 297 20.660 2.777 24.904 1.00 28.08 ATOM 1062 CB ASP 297 21.555 2.552 26.129 1.00 51.15 ATOM 1063 CG ASP 29722.207 3.835 26.629 1.00 51.15 ATOM 1064 OD1 ASP 297 22.508 4.725 25.804 1.00 51.15 ATOM 1065 OD2 ASP 297 22.425 3.948 27.855 1.00 51.15 ATOM 1066 C ASP 297 20.079 1.450 24.434 1.00 28.08 ATOM 1067 O ASP 297 20.549 0.869 23.456 1.00 51.15 ATOM1068 N ALA 298 19.024 1.006 25.111 1.00 26.12 ATOM 1069 CA ALA 298 18.357 -0.245 24.778 1.00 26.12 ATOM 1070 CB ALA 298 17.253 -0.530 25.787 1.00 18.80 ATOM 1071 C ALA 298 17.790 -0.223 23.356 1.00 26.12 ATOM 1072 O ALA 298 18.014 -1.154 22.5751.00 18.80 ATOM 1073 N ILE 299 17.078 0.848 23.013 1.00 17.42 ATOM 1074 CA ILE 299 16.483 0.979 21.686 1.00 17.42 ATOM 1075 CB ILE 299 15.559 2.211 21.597 1.00 16.69 ATOM 1076 CG2 ILE 299 14.845 2.238 20.253 1.00 16.69 ATOM 1077 CG1 ILE 29914.515 2.149 22.712 1.00 16.69 ATOM 1078 CD1 ILE 299 13.713 3.406 22.872 1.00 16.69 ATOM 1079 C ILE 299 17.563 1.042 20.609 1.00 17.42 ATOM 1080 O ILE 299 17.416 0.443 19.542 1.00 16.69 ATOM 1081 N PHE 300 18.652 1.752 20.889 1.00 14.46 ATOM1082 CA PHE 300 19.751 1.851 19.935 1.00 14.46 ATOM 1083 CB PHE 300 20.804 2.854 20.409 1.00 24.01 ATOM 1084 CG PHE 300 20.656 4.221 19.801 1.00 24.01 ATOM 1085 CD1 PHE 300 19.904 5.204 20.435 1.00 24.01 ATOM 1086 CD2 PHE 300 21.271 4.526 18.5911.00 24.01 ATOM 1087 CE1 PHE 300 19.766 6.472 19.873 1.00 24.01 ATOM 1088 CE2 PHE 300 21.140 5.791 18.020 1.00 24.01 ATOM 1089 CZ PHE 300 20.385 6.765 18.663 1.00 24.01 ATOM 1090 C PHE 300 20.383 0.480 19.726 1.00 14.46 ATOM 1091 O PHE 30020.696 0.102 18.596 1.00 24.01 ATOM 1092 N GLU 301 20.547 -0.270 20.813 1.00 21.61 ATOM 1093 CA GLU 301 21.123 -1.609 20.744 1.00 21.61 ATOM 1094 CB GLU 301 21.289 -2.192 22.143 1.00 23.89 ATOM 1095 C GLU 301 20.211 -2.498 19.904 1.00 21.61 ATOM 1096 O GLU 301 20.681 -3.251 19.043 1.00 23.89 ATOM 1097 N LEU 302 18.906 -2.390 20.140 1.00 14.43 ATOM 1098 CA LEU 302 17.922 -3.168 19.399 1.00 14.43 ATOM 1099 CB LEU 302 16.512 -2.872 19.912 1.00 23.43 ATOM 1100 CG LEU 302 15.350 -3.66919.312 1.00 23.43 ATOM 1101 CD1 LEU 302 15.459 -5.140 19.688 1.00 23.43 ATOM 1102 CD2 LEU 302 14.035 -3.094 19.804 1.00 23.43 ATOM 1103 C LEU 302 18.027 -2.812 17.917 1.00 14.43 ATOM 1104 O LEU 302 18.089 -3.697 17.066 1.00 23.43 ATOM 1105 NGLY 303 18.098 -1.515 17.625 1.00 15.17 ATOM 1106 CA GLY 303 18.208 -1.056 16.251 1.00 15.17 ATOM 1107 C GLY 303 19.411 -1.640 15.530 1.00 15.17 ATOM 1108 O GLY 303 19.290 -2.137 14.406 1.00 27.67

ATOM 1109 N LYS 304 20.570 -1.594 16.182 1.00 19.04 ATOM 1110 CA LYS 304 21.802 -2.127 15.605 1.00 19.04 ATOM 1111 CB LYS 304 22.979 -1.975 16.577 1.00 56.94 ATOM 1112 CG LYS 304 23.496 -0.556 16.741 1.00 56.94 ATOM 1113 CD LYS 304 24.811-0.524 17.516 1.00 56.94 ATOM 1114 CE LYS 304 24.634 -0.965 18.968 1.00 56.94 ATOM 1115 NZ LYS 304 23.838 0.008 19.778 1.00 56.94 ATOM 1116 C LYS 304 21.653 -3.596 15.229 1.00 19.04 ATOM 1117 O LYS 304 21.974 -3.993 14.107 1.00 56.94 ATOM 1118N SER 305 21.146 -4.394 16.164 1.00 24.46 ATOM 1119 CA SER 305 20.965 -5.822 15.932 1.00 24.46 ATOM 1120 CB SER 305 20.610 -6.533 17.240 1.00 37.46 ATOM 1121 OG SER 305 19.444 -5.984 17.827 1.00 37.46 ATOM 1122 C SER 305 19.926 -6.128 14.853 1.00 24.46 ATOM 1123 O SER 305 20.146 -6.996 14.006 1.00 37.46 ATOM 1124 N LEU 306 18.819 -5.390 14.858 1.00 25.47 ATOM 1125 CA LEU 306 17.753 -5.592 13.881 1.00 25.47 ATOM 1126 CB LEU 306 16.525 -4.746 14.224 1.00 15.99 ATOM 1127 CG LEU 306 15.700-5.190 15.432 1.00 15.99 ATOM 1128 CD1 LEU 306 14.504 -4.271 15.600 1.00 15.99 ATOM 1129 CD2 LEU 306 15.244 -6.624 15.247 1.00 15.99 ATOM 1130 C LEU 306 18.174 -5.330 12.439 1.00 25.47 ATOM 1131 O LEU 306 17.596 -5.902 11.513 1.00 15.99 ATOM1132 N SER 307 19.182 -4.482 12.247 1.00 24.28 ATOM 1133 CA SER 307 19.670 -4.160 10.907 1.00 24.28 ATOM 1134 CB SER 307 20.910 -3.263 10.989 1.00 40.92 ATOM 1135 OG SER 307 20.617 -2.028 11.622 1.00 40.92 ATOM 1136 C SER 307 19.995 -5.422 10.1071.00 24.28 ATOM 1137 O SER 307 19.625 -5.535 8.936 1.00 40.92 ATOM 1138 N ALA 308 20.644 -6.383 10.761 1.00 30.97 ATOM 1139 CA ALA 308 21.027 -7.640 10.124 1.00 30.97 ATOM 1140 CB ALA 308 22.004 -8.399 11.013 1.00 37.84 ATOM 1141 C ALA 30819.830 -8.528 9.779 1.00 30.97 ATOM 1142 O ALA 308 19.897 -9.336 8.853 1.00 37.84 ATOM 1143 N PHE 309 18.737 -8.372 10.520 1.00 22.78 ATOM 1144 CA PHE 309 17.533 -9.166 10.292 1.00 22.78 ATOM 1145 CB PHE 309 16.571 -9.037 11.477 1.00 30.14 ATOM1146 CG PHE 309 17.032 -9.751 12.716 1.00 30.14 ATOM 1147 CD1 PHE 309 16.299 -10.809 13.236 1.00 30.14 ATOM 1148 CD2 PHE 309 18.204 -9.372 13.359 1.00 30.14 ATOM 1149 CE1 PHE 309 16.725 -11.481 14.378 1.00 30.14 ATOM 1150 CE2 PHE 309 18.640-10.038 14.503 1.00 30.14 ATOM 1151 CZ PHE 309 17.896 -11.094 15.013 1.00 30.14 ATOM 1152 C PHE 309 16.818 -8.813 8.990 1.00 22.78 ATOM 1153 O PHE 309 16.068 -9.631 8.451 1.00 30.14 ATOM 1154 N ASN 310 17.051 -7.598 8.496 1.00 35.30 ATOM 1155CA ASN 310 16.441 -7.109 7.255 1.00 35.30 ATOM 1156 CB ASN 310 17.109 -7.760 6.037 1.00 28.28 ATOM 1157 C ASN 310 14.929 -7.339 7.229 1.00 35.30 ATOM 1158 O ASN 310 14.395 -7.970 6.312 1.00 28.28 ATOM 1159 N LEU 311 14.249 -6.831 8.251 1.00 27.52 ATOM 1160 CA LEU 311 12.803 -6.979 8.369 1.00 27.52 ATOM 1161 CB LEU 311 12.351 -6.630 9.788 1.00 22.62 ATOM 1162 CG LEU 311 12.950 -7.396 10.968 1.00 22.62 ATOM 1163 CD1 LEU 311 12.360 -6.864 12.268 1.00 22.62 ATOM 1164 CD2 LEU 311 12.672-8.881 10.821 1.00 22.62 ATOM 1165 C LEU 311 12.060 -6.085 7.382 1.00 27.52 ATOM 1166 O LEU 311 12.519 -4.986 7.067 1.00 22.62 ATOM 1167 N ASP 312 10.918 -6.563 6.892 1.00 16.74 ATOM 1168 CA ASP 312 10.095 -5.789 5.968 1.00 16.74 ATOM 1169 CBASP 312 9.803 -6.578 4.673 1.00 16.35 ATOM 1170 CG ASP 312 8.924 -7.814 4.888 1.00 16.35 ATOM 1171 OD1 ASP 312 8.591 -8.168 6.037 1.00 16.35 ATOM 1172 OD2 ASP 312 8.559 -8.446 3.876 1.00 16.35 ATOM 1173 C ASP 312 8.808 -5.354 6.678 1.00 16.74 ATOM 1174 O ASP 312 8.535 -5.798 7.797 1.00 16.35 ATOM 1175 N ASP 313 8.007 -4.520 6.019 1.00 5.43 ATOM 1176 CA ASP 313 6.758 -4.016 6.592 1.00 5.43 ATOM 1177 CB ASP 313 5.974 -3.201 5.559 1.00 31.80 ATOM 1178 CG ASP 313 6.670 -1.906 5.183 1.00 31.80 ATOM 1179 OD1 ASP 313 7.392 -1.340 6.033 1.00 31.80 ATOM 1180 OD2 ASP 313 6.493 -1.452 4.032 1.00 31.80 ATOM 1181 C ASP 313 5.849 -5.081 7.189 1.00 5.43 ATOM 1182 O ASP 313 5.216 -4.849 8.221 1.00 31.80 ATOM 1183 N THR 314 5.777 -6.2386.543 1.00 12.98 ATOM 1184 CA THR 314 4.934 -7.327 7.022 1.00 12.98 ATOM 1185 CB THR 314 4.825 -8.441 5.968 1.00 18.90 ATOM 1186 OG1 THR 314 4.249 -7.904 4.769 1.00 18.90 ATOM 1187 CG2 THR 314 3.960 -9.578 6.477 1.00 18.90 ATOM 1188 C THR 3145.426 -7.910 8.349 1.00 12.98 ATOM 1189 O THR 314 4.636 -8.124 9.268 1.00 18.90 ATOM 1190 N GLU 315 6.731 -8.135 8.457 1.00 9.13 ATOM 1191 CA GLU 315 7.316 -8.685 9.675 1.00 9.13 ATOM 1192 CB GLU 315 8.771 -9.078 9.427 1.00 11.49 ATOM 1193 CGGLU 315 8.870 -10.323 8.562 1.00 11.49 ATOM 1194 CD GLU 315 10.233 -10.544 7.945 1.00 11.49 ATOM 1195 OE1 GLU 315 10.964 -9.561 7.705 1.00 11.49 ATOM 1196 OE2 GLU 315 10.558 -11.715 7.669 1.00 11.49 ATOM 1197 C GLU 315 7.180 -7.720 10.847 1.00 9.13 ATOM 1198 O GLU 315 6.863 -8.131 11.967 1.00 11.49 ATOM 1199 N VAL 316 7.376 -6.433 10.575 1.00 9.46 ATOM 1200 CA VAL 316 7.240 -5.406 11.602 1.00 9.46 ATOM 1201 CB VAL 316 7.655 -4.015 11.063 1.00 7.95 ATOM 1202 CG1 VAL 316 7.434 -2.94112.124 1.00 7.95 ATOM 1203 CG2 VAL 316 9.112 -4.037 10.625 1.00 7.95 ATOM 1204 C VAL 316 5.777 -5.365 12.051 1.00 9.46 ATOM 1205 O VAL 316 5.484 -5.300 13.247 1.00 7.95 ATOM 1206 N ALA 317 4.866 -5.438 11.083 1.00 5.52 ATOM 1207 CA ALA 3173.434 -5.417 11.355 1.00 5.52 ATOM 1208 CB ALA 317 2.656 -5.415 10.054 1.00 10.98 ATOM 1209 C ALA 317 3.002 -6.595 12.225 1.00 5.52 ATOM 1210 O ALA 317 2.317 -6.412 13.230 1.00 10.98 ATOM 1211 N LEU 318 3.411 -7.799 11.838 1.00 8.62 ATOM 1212CA LEU 318 3.067 -9.003 12.584 1.00 8.62 ATOM 1213 CB LEU 318 3.523 -10.249 11.825 1.00 10.49 ATOM 1214 CG LEU 318 2.770 -10.494 10.514 1.00 10.49 ATOM 1215 CD1 LEU 318 3.376 -11.664 9.769 1.00 10.49 ATOM 1216 CD2 LEU 318 1.297 -10.741 10.7991.00 10.49 ATOM 1217 C LEU 318 3.674 -8.971 13.978 1.00 8.62 ATOM 1218 O LEU 318 3.047 -9.407 14.945 1.00 10.49 ATOM 1219 N LEU 319 4.885 -8.435 14.082 1.00 9.43 ATOM 1220 CA LEU 319 5.560 -8.325 15.366 1.00 9.43 ATOM 1221 CB LEU 319 6.975-7.773 15.173 1.00 24.05 ATOM 1222 CG LEU 319 7.901 -7.680 16.389 1.00 24.05 ATOM 1223 CD1 LEU 319 7.889 -8.977 17.182 1.00 24.05 ATOM 1224 CD2 LEU 319 9.310 -7.356 15.922 1.00 24.05 ATOM 1225 C LEU 319 4.731 -7.404 16.259 1.00 9.43 ATOM 1226 OLEU 319 4.456 -7.731 17.416 1.00 24.05 ATOM 1227 N GLN 320 4.287 -6.282 15.699 1.00 8.67 ATOM 1228 CA GLN 320 3.467 -5.325 16.437 1.00 8.67 ATOM 1229 CB GLN 320 3.151 -4.102 15.573 1.00 10.94 ATOM 1230 CG GLN 320 4.361 -3.256 15.218 1.00 10.94 ATOM 1231 CD GLN 320 4.025 -2.045 14.359 1.00 10.94 ATOM 1232 OE1 GLN 320 4.889 -1.217 14.082 1.00 10.94 ATOM 1233 NE2 GLN 320 2.773 -1.940 13.924 1.00 10.94 ATOM 1234 C GLN 320 2.169 -5.984 16.895 1.00

8.67 ATOM 1235 O GLN 320 1.708 -5.751 18.013 1.00 10.94 ATOM 1236 N ALA 321 1.586 -6.806 16.028 1.00 9.21 ATOM 1237 CA ALA 321 0.349 -7.513 16.342 1.00 9.21 ATOM 1238 CB ALA 321 -0.136 -8.283 15.129 1.00 12.83 ATOM 1239 C ALA 321 0.558-8.460 17.523 1.00 9.21 ATOM 1240 O ALA 321 -0.315 -8.591 18.382 1.00 12.83 ATOM 1241 N VAL 322 1.718 -9.111 17.566 1.00 9.10 ATOM 1242 CA VAL 322 2.043 -10.030 18.651 1.00 9.10 ATOM 1243 CB VAL 322 3.340 -10.827 18.352 1.00 15.92 ATOM 1244 CG1VAL 322 3.783 -11.614 19.575 1.00 15.92 ATOM 1245 CG2 VAL 322 3.106 -11.780 17.194 1.00 15.92 ATOM 1246 C VAL 322 2.192 -9.256 19.960 1.00 9.10 ATOM 1247 O VAL 322 1.707 -9.691 21.003 1.00 15.92 ATOM 1248 N LEU 323 2.856 -8.106 19.893 1.00 11.07 ATOM 1249 CA LEU 323 3.062 -7.257 21.064 1.00 11.07 ATOM 1250 CB LEU 323 3.959 -6.070 20.705 1.00 16.31 ATOM 1251 CG LEU 323 5.377 -6.393 20.229 1.00 16.31 ATOM 1252 CD1 LEU 323 6.039 -5.149 19.669 1.00 16.31 ATOM 1253 CD2 LEU 323 6.187 -6.96621.375 1.00 16.31 ATOM 1254 C LEU 323 1.729 -6.742 21.595 1.00 11.07 ATOM 1255 O LEU 323 1.523 -6.650 22.803 1.00 16.31 ATOM 1256 N LEU 324 0.827 -6.413 20.677 1.00 13.48 ATOM 1257 CA LEU 324 -0.494 -5.900 21.015 1.00 13.48 ATOM 1258 CB LEU 324-1.185 -5.383 19.752 1.00 15.92 ATOM 1259 CG LEU 324 -2.607 -4.837 19.889 1.00 15.92 ATOM 1260 CD1 LEU 324 -2.602 -3.547 20.692 1.00 15.92 ATOM 1261 CD2 LEU 324 -3.182 -4.598 18.511 1.00 15.92 ATOM 1262 C LEU 324 -1.393 -6.924 21.707 1.00 13.48 ATOM 1263 O LEU 324 -1.896 -6.678 22.802 1.00 15.92 ATOM 1264 N MET 325 -1.593 -8.074 21.072 1.00 11.47 ATOM 1265 CA MET 325 -2.458 -9.111 21.631 1.00 11.47 ATOM 1266 CB MET 325 -2.959 -10.043 20.520 1.00 22.90 ATOM 1267 CG MET 325 -3.689 -9.34719.375 1.00 22.90 ATOM 1268 SD MET 325 -5.052 -8.287 19.908 1.00 22.90 ATOM 1269 CE MET 325 -6.284 -9.475 20.353 1.00 22.90 ATOM 1270 C MET 325 -1.814 -9.932 22.752 1.00 11.47 ATOM 1271 O MET 325 -1.899 -11.160 22.758 1.00 22.90 ATOM 1272 N SER326 -1.193 -9.256 23.711 1.00 30.07 ATOM 1273 CA SER 326 -0.543 -9.936 24.826 1.00 30.07 ATOM 1274 CB SER 326 0.723 -9.175 25.239 1.00 32.79 ATOM 1275 OG SER 326 1.283 -9.699 26.433 1.00 32.79 ATOM 1276 C SER 326 -1.492 -10.061 26.014 1.00 30.07 ATOM 1277 O SER 326 -2.343 -9.198 26.235 1.00 32.79 ATOM 1278 N THR 327 -1.347 -11.143 26.773 1.00 29.08 ATOM 1279 CA THR 327 -2.179 -11.368 27.948 1.00 29.08 ATOM 1280 CB THR 327 -2.705 -12.817 27.998 1.00 36.96 ATOM 1281 OG1 THR 327 -1.612-13.734 27.856 1.00 36.96 ATOM 1282 CG2 THR 327 -3.716 -13.055 26.890 1.00 36.96 ATOM 1283 C THR 327 -1.426 -11.049 29.239 1.00 29.08 ATOM 1284 O THR 327 -1.930 -11.295 30.333 1.00 36.96 ATOM 1285 N ASP 328 -0.214 -10.513 29.111 1.00 38.93 ATOM1286 CA ASP 328 0.596 -10.152 30.273 1.00 38.93 ATOM 1287 CB ASP 328 2.082 -10.089 29.899 1.00 85.70 ATOM 1288 CG ASP 328 2.660 -11.451 29.556 1.00 85.70 ATOM 1289 OD1 ASP 328 3.388 -11.554 28.542 1.00 85.70 ATOM 1290 OD2 ASP 328 2.393 -12.41830.303 1.00 85.70 ATOM 1291 C ASP 328 0.148 -8.810 30.845 1.00 38.93 ATOM 1292 O ASP 328 0.962 -7.911 31.061 1.00 85.70 ATOM 1293 N ARG 329 -1.154 -8.673 31.070 1.00 28.95 ATOM 1294 CA ARG 329 -1.716 -7.445 31.608 1.00 28.95 ATOM 1295 CB ARG329 -2.390 -6.612 30.509 1.00 38.88 ATOM 1296 CG ARG 329 -1.449 -5.887 29.554 1.00 38.88 ATOM 1297 CD ARG 329 -1.107 -6.739 28.347 1.00 38.88 ATOM 1298 NE ARG 329 -0.322 -6.005 27.356 1.00 38.88 ATOM 1299 CZ ARG 329 1.006 -5.936 27.351 1.00 38.88 ATOM 1300 NH1 ARG 329 1.713 -6.552 28.290 1.00 38.88 ATOM 1301 NH2 ARG 329 1.631 -5.270 26.391 1.00 38.88 ATOM 1302 C ARG 329 -2.745 -7.790 32.672 1.00 28.95 ATOM 1303 O ARG 329 -3.279 -8.898 32.696 1.00 38.88 ATOM 1304 N SER 330 -3.029-6.829 33.542 1.00 42.07 ATOM 1305 CA SER 330 -3.999 -7.025 34.607 1.00 42.07 ATOM 1306 CB SER 330 -3.488 -6.399 35.899 1.00 37.35 ATOM 1307 C SER 330 -5.340 -6.413 34.220 1.00 42.07 ATOM 1308 O SER 330 -5.386 -5.382 33.550 1.00 37.35 ATOM 1309N GLY 331 -6.424 -7.085 34.598 1.00 26.57 ATOM 1310 CA GLY 331 -7.754 -6.572 34.318 1.00 26.57 ATOM 1311 C GLY 331 -8.404 -6.915 32.991 1.00 26.57 ATOM 1312 O GLY 331 -9.462 -6.371 32.671 1.00 30.06 ATOM 1313 N LEU 332 -7.797 -7.807 32.214 1.00 31.47 ATOM 1314 CA LEU 332 -8.374 -8.189 30.928 1.00 31.47 ATOM 1315 CB LEU 332 -7.351 -8.933 30.065 1.00 23.83 ATOM 1316 CG LEU 332 -6.261 -8.076 29.425 1.00 23.83 ATOM 1317 CD1 LEU 332 -5.296 -8.960 28.652 1.00 23.83 ATOM 1318 CD2 LEU 332-6.897 -7.041 28.509 1.00 23.83 ATOM 1319 C LEU 332 -9.630 -9.039 31.091 1.00 31.47 ATOM 1320 O LEU 332 -9.665 -9.969 31.895 1.00 23.83 ATOM 1321 N LEU 333 -10.659 -8.702 30.321 1.00 27.66 ATOM 1322 CA LEU 333 -11.927 -9.422 30.351 1.00 27.66 ATOM 1323 CB LEU 333 -13.072 -8.500 29.918 1.00 49.79 ATOM 1324 CG LEU 333 -13.416 -7.312 30.820 1.00 49.79 ATOM 1325 CD1 LEU 333 -14.328 -6.339 30.083 1.00 49.79 ATOM 1326 CD2 LEU 333 -14.072 -7.803 32.104 1.00 49.79 ATOM 1327 C LEU 333 -11.904-10.663 29.456 1.00 27.66 ATOM 1328 O LEU 333 -12.117 -11.780 29.919 1.00 49.79 ATOM 1329 N CYS 334 -11.616 -10.464 28.174 1.00 29.56 ATOM 1330 CA CYS 334 -11.583 -11.566 27.220 1.00 29.56 ATOM 1331 CB CYS 334 -12.134 -11.106 25.865 1.00 47.01 ATOM 1332 SG CYS 334 -13.888 -10.657 25.883 1.00 47.01 ATOM 1333 C CYS 334 -10.187 -12.161 27.050 1.00 29.56 ATOM 1334 O CYS 334 -9.652 -12.202 25.942 1.00 47.01 ATOM 1335 N VAL 335 -9.617 -12.655 28.147 1.00 30.69 ATOM 1336 CA VAL 335 -8.280-13.250 28.132 1.00 30.69 ATOM 1337 CB VAL 335 -7.913 -13.844 29.514 1.00 32.18 ATOM 1338 CG1 VAL 335 -6.517 -14.456 29.480 1.00 32.18 ATOM 1339 CG2 VAL 335 -7.988 -12.768 30.584 1.00 32.18 ATOM 1340 C VAL 335 -8.120 -14.340 27.068 1.00 30.69 ATOM 1341 O VAL 335 -7.149 -14.337 26.309 1.00 32.18 ATOM 1342 N ASP 336 -9.079 -15.260 27.012 1.00 30.13 ATOM 1343 CA ASP 336 -9.040 -16.360 26.052 1.00 30.13 ATOM 1344 CB ASP 336 -10.218 -17.311 26.284 1.00 63.22 ATOM 1345 CG ASP 336 -10.178-18.528 25.370 1.00 63.22 ATOM 1346 OD1 ASP 336 -11.119 -18.700 24.565 1.00 63.22 ATOM 1347 OD2 ASP 336 -9.205 -19.311 25.452 1.00 63.22 ATOM 1348 C ASP 336 -9.012 -15.903 24.594 1.00 30.13 ATOM 1349 O ASP 336 -8.156 -16.339 23.823 1.00 63.22 ATOM 1350 N LYS 337 -9.944 -15.027 24.223 1.00 26.63 ATOM 1351 CA LYS 337 -10.024 -14.515 22.856 1.00 26.63 ATOM 1352 CB LYS 337 -11.172 -13.516 22.729 1.00 21.38 ATOM 1353 C LYS 337 -8.706 -13.865 22.438 1.00 26.63 ATOM 1354 O LYS 337 -8.204-14.110 21.338 1.00 21.38 ATOM 1355 N ILE 338 -8.141 -13.060 23.334 1.00 24.65 ATOM 1356 CA ILE 338 -6.879 -12.376 23.078 1.00 24.65 ATOM 1357 CB ILE 338 -6.543 -11.380 24.215 1.00 20.45 ATOM 1358 CG2 ILE 338 -5.198 -10.719 23.966 1.00 20.45 ATOM 1359 CG1 ILE 338 -7.632 -10.308 24.308 1.00 20.45

ATOM 1360 CD1 ILE 338 -7.479 -9.374 25.486 1.00 20.45 ATOM 1361 C ILE 338 -5.744 -13.388 22.911 1.00 24.65 ATOM 1362 O ILE 338 -4.948 -13.288 21.974 1.00 20.45 ATOM 1363 N GLU 339 -5.700 -14.383 23.795 1.00 35.34 ATOM 1364 CA GLU 339-4.673 -15.422 23.745 1.00 35.34 ATOM 1365 CB GLU 339 -4.836 -16.388 24.916 1.00 29.51 ATOM 1366 C GLU 339 -4.744 -16.180 22.421 1.00 35.34 ATOM 1367 O GLU 339 -3.720 -16.421 21.777 1.00 29.51 ATOM 1368 N LYS 340 -5.959 -16.536 22.009 1.00 24.19 ATOM 1369 CA LYS 340 -6.168 -17.256 20.755 1.00 24.19 ATOM 1370 CB LYS 340 -7.627 -17.671 20.624 1.00 23.97 ATOM 1371 C LYS 340 -5.754 -16.377 19.576 1.00 24.19 ATOM 1372 O LYS 340 -5.197 -16.860 18.586 1.00 23.97 ATOM 1373 N SER 341 -6.000-15.079 19.708 1.00 16.85 ATOM 1374 CA SER 341 -5.651 -14.115 18.676 1.00 16.85 ATOM 1375 CB SER 341 -6.223 -12.744 19.033 1.00 26.59 ATOM 1376 OG SER 341 -5.852 -11.765 18.080 1.00 26.59 ATOM 1377 C SER 341 -4.137 -14.026 18.500 1.00 16.85 ATOM 1378 O SER 341 -3.638 -14.042 17.374 1.00 26.59 ATOM 1379 N GLN 342 -3.406 -13.932 19.608 1.00 17.35 ATOM 1380 CA GLN 342 -1.952 -13.845 19.537 1.00 17.35 ATOM 1381 CB GLN 342 -1.337 -13.597 20.913 1.00 30.07 ATOM 1382 CG GLN 342 0.140-13.245 20.832 1.00 30.07 ATOM 1383 CD GLN 342 0.811 -13.196 22.182 1.00 30.07 ATOM 1384 OE1 GLN 342 0.884 -14.201 22.884 1.00 30.07 ATOM 1385 NE2 GLN 342 1.318 -12.030 22.548 1.00 30.07 ATOM 1386 C GLN 342 -1.368 -15.118 18.944 1.00 17.35 ATOM1387 O GLN 342 -0.405 -15.066 18.178 1.00 30.07 ATOM 1388 N GLU 343 -1.949 -16.260 19.303 1.00 18.35 ATOM 1389 CA GLU 343 -1.489 -17.546 18.791 1.00 18.35 ATOM 1390 CB GLU 343 -2.308 -18.676 19.394 1.00 16.98 ATOM 1391 C GLU 343 -1.603 -17.56017.267 1.00 18.35 ATOM 1392 O GLU 343 -0.699 -18.026 16.568 1.00 16.98 ATOM 1393 N ALA 344 -2.706 -17.017 16.761 1.00 14.83 ATOM 1394 CA ALA 344 -2.946 -16.948 15.324 1.00 14.83 ATOM 1395 CB ALA 344 -4.327 -16.376 15.049 1.00 19.42 ATOM 1396 CALA 344 -1.872 -16.102 14.640 1.00 14.83 ATOM 1397 O ALA 344 -1.311 -16.507 13.619 1.00 19.42 ATOM 1398 N TYR 345 -1.586 -14.934 15.211 1.00 13.10 ATOM 1399 CA TYR 345 -0.569 -14.041 14.665 1.00 13.10 ATOM 1400 CB TYR 345 -0.573 -12.697 15.3931.00 2.00 ATOM 1401 CG TYR 345 -1.670 -11.767 14.938 1.00 2.00 ATOM 1402 CD1 TYR 345 -2.707 -11.409 15.794 1.00 2.00 ATOM 1403 CE1 TYR 345 -3.722 -10.562 15.377 1.00 2.00 ATOM 1404 CD2 TYR 345 -1.674 -11.248 13.647 1.00 2.00 ATOM 1405 CE2 TYR345 -2.683 -10.398 13.219 1.00 2.00 ATOM 1406 CZ TYR 345 -3.706 -10.061 14.087 1.00 2.00 ATOM 1407 OH TYR 345 -4.722 -9.233 13.669 1.00 2.00 ATOM 1408 C TYR 345 0.818 -14.666 14.732 1.00 13.10 ATOM 1409 O TYR 345 1.614 -14.504 13.811 1.00 2.00 ATOM 1410 N LEU 346 1.101 -15.387 15.813 1.00 12.59 ATOM 1411 CA LEU 346 2.396 -16.041 15.976 1.00 12.59 ATOM 1412 CB LEU 346 2.498 -16.715 17.347 1.00 22.61 ATOM 1413 CG LEU 346 2.899 -15.799 18.504 1.00 22.61 ATOM 1414 CD1 LEU 346 2.717 -16.51119.830 1.00 22.61 ATOM 1415 CD2 LEU 346 4.341 -15.357 18.324 1.00 22.61 ATOM 1416 C LEU 346 2.629 -17.057 14.865 1.00 12.59 ATOM 1417 O LEU 346 3.706 -17.099 14.272 1.00 22.61 ATOM 1418 N LEU 347 1.612 -17.862 14.574 1.00 18.42 ATOM 1419 CA LEU347 1.706 -18.863 13.517 1.00 18.42 ATOM 1420 CB LEU 347 0.471 -19.762 13.512 1.00 23.56 ATOM 1421 CG LEU 347 0.509 -20.965 14.456 1.00 23.56 ATOM 1422 CD1 LEU 347 -0.819 -21.702 14.398 1.00 23.56 ATOM 1423 CD2 LEU 347 1.659 -21.890 14.068 1.00 23.56 ATOM 1424 C LEU 347 1.870 -18.201 12.154 1.00 18.42 ATOM 1425 O LEU 347 2.672 -18.651 11.330 1.00 23.56 ATOM 1426 N ALA 348 1.099 -17.144 11.917 1.00 12.49 ATOM 1427 CA ALA 348 1.157 -16.403 10.663 1.00 12.49 ATOM 1428 CB ALA 348 0.098-15.302 10.654 1.00 14.77 ATOM 1429 C ALA 348 2.545 -15.798 10.504 1.00 12.49 ATOM 1430 O ALA 348 3.154 -15.874 9.436 1.00 14.77 ATOM 1431 N PHE 349 3.048 -15.246 11.602 1.00 15.52 ATOM 1432 CA PHE 349 4.357 -14.613 11.664 1.00 15.52 ATOM 1433CB PHE 349 4.566 -14.049 13.076 1.00 14.41 ATOM 1434 CG PHE 349 5.714 -13.085 13.203 1.00 14.41 ATOM 1435 CD1 PHE 349 6.473 -12.712 12.099 1.00 14.41 ATOM 1436 CD2 PHE 349 6.027 -12.540 14.443 1.00 14.41 ATOM 1437 CE1 PHE 349 7.523 -11.813 12.2301.00 14.41 ATOM 1438 CE2 PHE 349 7.075 -11.640 14.584 1.00 14.41 ATOM 1439 CZ PHE 349 7.825 -11.275 13.475 1.00 14.41 ATOM 1440 C PHE 349 5.444 -15.633 11.324 1.00 15.52 ATOM 1441 O PHE 349 6.252 -15.413 10.422 1.00 14.41 ATOM 1442 N GLU 3505.439 -16.760 12.026 1.00 13.20 ATOM 1443 CA GLU 350 6.424 -17.811 11.801 1.00 13.20 ATOM 1444 CB GLU 350 6.152 -18.995 12.734 1.00 33.43 ATOM 1445 CG GLU 350 7.068 -20.193 12.519 1.00 33.43 ATOM 1446 CD GLU 350 6.786 -21.331 13.482 1.00 33.43 ATOM 1447 OE1 GLU 350 7.746 -22.035 13.857 1.00 33.43 ATOM 1448 OE2 GLU 350 5.611 -21.525 13.865 1.00 33.43 ATOM 1449 C GLU 350 6.409 -18.283 10.352 1.00 13.20 ATOM 1450 O GLU 350 7.449 -18.355 9.694 1.00 33.43 ATOM 1451 N HIS 351 5.217 -18.5739.850 1.00 19.10 ATOM 1452 CA HIS 351 5.062 -19.051 8.485 1.00 19.10 ATOM 1453 CB HIS 351 3.632 -19.536 8.256 1.00 18.97 ATOM 1454 CG HIS 351 3.249 -20.700 9.117 1.00 18.97 ATOM 1455 CD2 HIS 351 3.987 -21.474 9.948 1.00 18.97 ATOM 1456 ND1 HIS351 1.960 -21.180 9.194 1.00 18.97 ATOM 1457 CE1 HIS 351 1.918 -22.195 10.039 1.00 18.97 ATOM 1458 NE2 HIS 351 3.134 -22.394 10.509 1.00 18.97 ATOM 1459 C HIS 351 5.477 -18.011 7.449 1.00 19.10 ATOM 1460 O HIS 351 5.955 -18.366 6.371 1.00 18.97 ATOM 1461 N TYR 352 5.304 -16.732 7.767 1.00 9.38 ATOM 1462 CA TYR 352 5.711 -15.683 6.843 1.00 9.38 ATOM 1463 CB TYR 352 5.168 -14.317 7.257 1.00 16.06 ATOM 1464 CG TYR 352 5.539 -13.238 6.268 1.00 16.06 ATOM 1465 CD1 TYR 352 4.939 -13.190 5.0081.00 16.06 ATOM 1466 CE1 TYR 352 5.321 -12.242 4.060 1.00 16.06 ATOM 1467 CD2 TYR 352 6.531 -12.303 6.562 1.00 16.06 ATOM 1468 CE2 TYR 352 6.923 -11.349 5.620 1.00 16.06 ATOM 1469 CZ TYR 352 6.313 -11.326 4.371 1.00 16.06 ATOM 1470 OH TYR 3526.710 -10.401 3.431 1.00 16.06 ATOM 1471 C TYR 352 7.234 -15.639 6.812 1.00 9.38 ATOM 1472 O TYR 352 7.838 -15.475 5.751 1.00 16.06 ATOM 1473 N VAL 353 7.851 -15.789 7.980 1.00 15.38 ATOM 1474 CA VAL 353 9.305 -15.790 8.087 1.00 15.38 ATOM 1475CB VAL 353 9.761 -15.945 9.558 1.00 18.40 ATOM 1476 CG1 VAL 353 11.262 -16.163 9.633 1.00 18.40 ATOM 1477 CG2 VAL 353 9.384 -14.703 10.349 1.00 18.40 ATOM 1478 C VAL 353 9.853 -16.938 7.237 1.00 15.38 ATOM 1479 O VAL 353 10.850 -16.773 6.525 1.00 18.40 ATOM 1480 N ASN 354 9.183 -18.086 7.298 1.00 14.74 ATOM 1481 CA ASN 354 9.578 -19.259 6.521 1.00 14.74 ATOM 1482 CB ASN 354 8.640 -20.435 6.799 1.00 19.97 ATOM 1483 CG ASN 354 8.832 -21.020 8.180 1.00 19.97 ATOM 1484 OD1 ASN 354 9.879-20.848 8.799 1.00 19.97 ATOM 1485 ND2 ASN 354 7.826 -21.734 8.664 1.00

19.97 ATOM 1486 C ASN 354 9.550 -18.939 5.034 1.00 14.74 ATOM 1487 O ASN 354 10.452 -19.319 4.290 1.00 19.97 ATOM 1488 N HIS 355 8.507 -18.230 4.613 1.00 13.03 ATOM 1489 CA HIS 355 8.329 -17.837 3.220 1.00 13.03 ATOM 1490 CB HIS 3556.960 -17.164 3.042 1.00 24.39 ATOM 1491 CG HIS 355 6.753 -16.541 1.695 1.00 24.39 ATOM 1492 CD2 HIS 355 7.195 -15.370 1.176 1.00 24.39 ATOM 1493 ND1 HIS 355 6.009 -17.138 0.701 1.00 24.39 ATOM 1494 CE1 HIS 355 6.005 -16.368 -0.372 1.00 24.39 ATOM 1495 NE2 HIS 355 6.720 -15.289 -0.107 1.00 24.39 ATOM 1496 C HIS 355 9.434 -16.894 2.758 1.00 13.03 ATOM 1497 O HIS 355 9.834 -16.920 1.595 1.00 24.39 ATOM 1498 N ARG 356 9.878 -16.027 3.660 1.00 19.55 ATOM 1499 CA ARG 356 10.920 -15.0543.358 1.00 19.55 ATOM 1500 CB ARG 356 10.970 -14.001 4.460 1.00 22.01 ATOM 1501 CG ARG 356 9.772 -13.081 4.454 1.00 22.01 ATOM 1502 CD ARG 356 10.097 -11.784 3.750 1.00 22.01 ATOM 1503 NE ARG 356 10.932 -10.934 4.592 1.00 22.01 ATOM 1504 CZ ARG356 11.822 -10.059 4.137 1.00 22.01 ATOM 1505 NH1 ARG 356 12.010 -9.907 2.833 1.00 22.01 ATOM 1506 NH2 ARG 356 12.519 -9.325 4.992 1.00 22.01 ATOM 1507 C ARG 356 12.297 -15.675 3.158 1.00 19.55 ATOM 1508 O ARG 356 13.127 -15.126 2.434 1.00 22.01 ATOM 1509 N LYS 357 12.547 -16.788 3.841 1.00 23.18 ATOM 1510 CA LYS 357 13.815 -17.504 3.739 1.00 23.18 ATOM 1511 CB LYS 357 13.879 -18.273 2.415 1.00 42.91 ATOM 1512 CG LYS 357 12.750 -19.277 2.274 1.00 42.91 ATOM 1513 CD LYS 357 12.773 -20.0210.960 1.00 42.91 ATOM 1514 CE LYS 357 11.619 -21.011 0.913 1.00 42.91 ATOM 1515 NZ LYS 357 11.629 -21.845 -0.316 1.00 42.91 ATOM 1516 C LYS 357 15.047 -16.619 3.918 1.00 23.18 ATOM 1517 O LYS 357 15.816 -16.396 2.982 1.00 42.91 ATOM 1518 N HIS358 15.228 -16.122 5.137 1.00 32.39 ATOM 1519 CA HIS 358 16.367 -15.272 5.460 1.00 32.39 ATOM 1520 CB HIS 358 16.181 -14.626 6.835 1.00 26.77 ATOM 1521 CG HIS 358 15.232 -13.468 6.841 1.00 26.77 ATOM 1522 CD2 HIS 358 15.452 -12.138 6.709 1.00 26.77 ATOM 1523 ND1 HIS 358 13.875 -13.615 7.028 1.00 26.77 ATOM 1524 CE1 HIS 358 13.300 -12.426 7.012 1.00 26.77 ATOM 1525 NE2 HIS 358 14.234 -11.513 6.821 1.00 26.77 ATOM 1526 C HIS 358 17.633 -16.115 5.480 1.00 32.39 ATOM 1527 O HIS 35817.618 -17.248 5.961 1.00 26.77 ATOM 1528 N ASN 359 18.728 -15.561 4.972 1.00 41.97 ATOM 1529 CA ASN 359 20.000 -16.273 4.959 1.00 41.97 ATOM 1530 CB ASN 359 20.909 -15.716 3.863 1.00 46.84 ATOM 1531 C ASN 359 20.663 -16.134 6.331 1.00 41.97 ATOM 1532 O ASN 359 21.821 -15.731 6.436 1.00 46.84 ATOM 1533 N ILE 360 19.908 -16.450 7.379 1.00 35.72 ATOM 1534 CA ILE 360 20.394 -16.359 8.753 1.00 35.72 ATOM 1535 CB ILE 360 19.819 -15.113 9.480 1.00 36.14 ATOM 1536 CG2 ILE 360 20.327 -15.05010.918 1.00 36.14 ATOM 1537 CG1 ILE 360 20.204 -13.833 8.734 1.00 36.14 ATOM 1538 CD1 ILE 360 19.526 -12.591 9.265 1.00 36.14 ATOM 1539 C ILE 360 19.935 -17.611 9.493 1.00 35.72 ATOM 1540 O ILE 360 18.748 -17.953 9.479 1.00 36.14 ATOM 1541 NPRO 361 20.877 -18.338 10.109 1.00 31.56 ATOM 1542 CD PRO 361 22.334 -18.114 10.100 1.00 33.50 ATOM 1543 CA PRO 361 20.532 -19.556 10.847 1.00 31.56 ATOM 1544 CB PRO 361 21.901 -20.163 11.161 1.00 33.50 ATOM 1545 CG PRO 361 22.801 -18.967 11.2491.00 33.50 ATOM 1546 C PRO 361 19.743 -19.256 12.121 1.00 31.56 ATOM 1547 O PRO 361 20.080 -18.338 12.867 1.00 33.50 ATOM 1548 N HIS 362 18.688 -20.034 12.355 1.00 18.84 ATOM 1549 CA HIS 362 17.840 -19.887 13.541 1.00 18.84 ATOM 1550 CB HIS 36218.656 -20.151 14.812 1.00 31.38 ATOM 1551 CG HIS 362 19.540 -21.357 14.731 1.00 31.38 ATOM 1552 CD2 HIS 362 19.250 -22.667 14.537 1.00 31.38 ATOM 1553 ND1 HIS 362 20.910 -21.286 14.860 1.00 31.38 ATOM 1554 CE1 HIS 362 21.427 -22.497 14.754 1.00 31.38 ATOM 1555 NE2 HIS 362 20.439 -23.353 14.558 1.00 31.38 ATOM 1556 C HIS 362 17.189 -18.506 13.628 1.00 18.84 ATOM 1557 O HIS 362 16.980 -17.979 14.723 1.00 31.38 ATOM 1558 N PHE 363 16.825 -17.950 12.476 1.00 18.69 ATOM 1559 CA PHE 36316.209 -16.630 12.408 1.00 18.69 ATOM 1560 CB PHE 363 15.825 -16.302 10.962 1.00 19.25 ATOM 1561 CG PHE 363 15.339 -14.894 10.765 1.00 19.25 ATOM 1562 CD1 PHE 363 16.239 -13.862 10.530 1.00 19.25 ATOM 1563 CD2 PHE 363 13.981 -14.598 10.819 1.00 19.25 ATOM 1564 CE1 PHE 363 15.794 -12.556 10.351 1.00 19.25 ATOM 1565 CE2 PHE 363 13.527 -13.296 10.642 1.00 19.25 ATOM 1566 CZ PHE 363 14.435 -12.273 10.407 1.00 19.25 ATOM 1567 C PHE 363 14.995 -16.461 13.323 1.00 18.69 ATOM 1568 O PHE 36314.955 -15.540 14.138 1.00 19.25 ATOM 1569 N TRP 364 14.016 -17.351 13.191 1.00 16.46 ATOM 1570 CA TRP 364 12.797 -17.280 13.995 1.00 16.46 ATOM 1571 CB TRP 364 11.882 -18.482 13.706 1.00 17.81 ATOM 1572 CG TRP 364 10.588 -18.488 14.481 1.00 17.81 ATOM 1573 CD2 TRP 364 9.586 -17.458 14.504 1.00 17.81 ATOM 1574 CE2 TRP 364 8.547 -17.905 15.350 1.00 17.81 ATOM 1575 CE3 TRP 364 9.467 -16.202 13.894 1.00 17.81 ATOM 1576 CD1 TRP 364 10.126 -19.486 15.290 1.00 17.81 ATOM 1577 NE1 TRP 3648.902 -19.144 15.814 1.00 17.81 ATOM 1578 CZ2 TRP 364 7.403 -17.142 15.602 1.00 17.81 ATOM 1579 CZ3 TRP 364 8.329 -15.444 14.145 1.00 17.81 ATOM 1580 CH2 TRP 364 7.312 -15.919 14.992 1.00 17.81 ATOM 1581 C TRP 364 13.046 -17.114 15.500 1.00 16.46 ATOM 1582 O TRP 364 12.595 -16.133 16.087 1.00 17.81 ATOM 1583 N PRO 365 13.779 -18.051 16.137 1.00 18.31 ATOM 1584 CD PRO 365 14.342 -19.314 15.625 1.00 25.61 ATOM 1585 CA PRO 365 14.038 -17.920 17.577 1.00 18.31 ATOM 1586 CB PRO 36514.939 -19.118 17.874 1.00 25.61 ATOM 1587 CG PRO 365 14.500 -20.130 16.882 1.00 25.61 ATOM 1588 C PRO 365 14.732 -16.606 17.933 1.00 18.31 ATOM 1589 O PRO 365 14.387 -15.963 18.926 1.00 25.61 ATOM 1590 N LYS 366 15.699 -16.207 17.112 1.00 25.16 ATOM 1591 CA LYS 366 16.439 -14.968 17.338 1.00 25.16 ATOM 1592 CB LYS 366 17.537 -14.805 16.289 1.00 40.51 ATOM 1593 CG LYS 366 18.679 -15.792 16.417 1.00 40.51 ATOM 1594 CD LYS 366 19.664 -15.607 15.278 1.00 40.51 ATOM 1595 CE LYS 366 20.884-16.492 15.440 1.00 40.51 ATOM 1596 NZ LYS 366 21.800 -16.360 14.275 1.00 40.51 ATOM 1597 C LYS 366 15.521 -13.747 17.317 1.00 25.16 ATOM 1598 O LYS 366 15.593 -12.893 18.202 1.00 49.51 ATOM 1599 N LEU 367 14.661 -13.666 16.307 1.00 25.30 ATOM1600 CA LEU 367 13.729 -12.551 16.184 1.00 25.30 ATOM 1601 CB LEU 367 12.989 -12.620 14.845 1.00 27.80 ATOM 1602 CG LEU 367 11.964 -11.519 14.561 1.00 27.80 ATOM 1603 CD1 LEU 367 12.621 -10.147 14.679 1.00 27.80 ATOM 1604 CD2 LEU 367 11.367-11.724 13.175 1.00 27.80 ATOM 1605 C LEU 367 12.730 -12.596 17.332 1.00 25.30 ATOM 1606 O LEU 367 12.337 -11.563 17.877 1.00 27.80 ATOM 1607 N LEU 368 12.345 -13.807 17.712 1.00 26.12 ATOM 1608 CA LEU 368 11.396 -14.019 18.793 1.00 26.12 ATOM1609 CB LEU 368 11.105 -15.515 18.919 1.00 33.27 ATOM 1610 CG LEU 368 9.696 -15.976 19.289 1.00 33.27

ATOM 1611 CD1 LEU 368 8.640 -15.182 18.529 1.00 33.27 ATOM 1612 CD2 LEU 368 9.582 -17.460 18.976 1.00 33.27 ATOM 1613 C LEU 368 11.973 -13.466 20.096 1.00 26.12 ATOM 1614 O LEU 368 11.249 -12.920 20.930 1.00 33.27 ATOM 1615 N MET 36913.289 -13.571 20.244 1.00 24.39 ATOM 1616 CA MET 369 13.971 -13.076 21.432 1.00 24.39 ATOM 1617 CB MET 369 15.382 -13.656 21.511 1.00 47.44 ATOM 1618 CG MET 369 15.407 -15.096 22.009 1.00 47.44 ATOM 1619 SD MET 369 16.850 -16.029 21.464 1.00 47.44 ATOM 1620 CE MET 369 18.186 -15.114 22.246 1.00 47.44 ATOM 1621 C MET 369 13.996 -11.552 21.491 1.00 24.39 ATOM 1622 O MET 369 14.212 -10.971 22.557 1.00 47.44 ATOM 1623 N LYS 370 13.749 -10.904 20.354 1.00 27.31 ATOM 1624 CA LYS 37013.713 -9.445 20.297 1.00 27.31 ATOM 1625 CB LYS 370 13.739 -8.951 18.847 1.00 28.20 ATOM 1626 CG LYS 370 15.004 -9.312 18.090 1.00 28.20 ATOM 1627 CD LYS 370 16.231 -8.810 18.824 1.00 28.20 ATOM 1628 CE LYS 370 17.512 -9.244 18.142 1.00 28.20 ATOM 1629 NZ LYS 370 18.696 -8.851 18.952 1.00 28.20 ATOM 1630 C LYS 370 12.453 -8.945 21.002 1.00 27.31 ATOM 1631 O LYS 370 12.424 -7.835 21.535 1.00 28.20 ATOM 1632 N VAL 371 11.413 -9.776 21.009 1.00 26.41 ATOM 1633 CA VAL 371 10.157 -9.43221.668 1.00 26.41 ATOM 1634 CB VAL 371 9.109 -10.561 21.512 1.00 25.61 ATOM 1635 CG1 VAL 371 7.825 -10.205 22.245 1.00 25.61 ATOM 1636 CG2 VAL 371 8.819 -10.805 20.044 1.00 25.61 ATOM 1637 C VAL 371 10.450 -9.205 23.151 1.00 26.41 ATOM 1638 OVAL 371 9.962 -8.248 23.752 1.00 25.61 ATOM 1639 N THR 372 11.294 -10.065 23.713 1.00 26.28 ATOM 1640 CA THR 372 11.683 -9.972 25.116 1.00 26.28 ATOM 1641 CB THR 372 12.656 -11.109 25.500 1.00 28.14 ATOM 1642 OG1 THR 372 12.025 -12.377 25.2751.00 28.14 ATOM 1643 CG2 THR 372 13.055 -11.001 26.965 1.00 28.14 ATOM 1644 C THR 372 12.358 -8.624 25.372 1.00 26.28 ATOM 1645 O THR 372 12.047 -7.937 26.350 1.00 28.14 ATOM 1646 N ASP 373 13.269 -8.247 24.478 1.00 15.09 ATOM 1647 CA ASP 37313.977 -6.979 24.588 1.00 15.09 ATOM 1648 CB ASP 373 14.976 -6.822 23.435 1.00 37.94 ATOM 1649 CG ASP 373 16.065 -7.893 23.445 1.00 37.94 ATOM 1650 OD1 ASP 373 16.248 -8.571 24.483 1.00 37.94 ATOM 1651 OD2 ASP 373 16.750 -8.052 22.410 1.00 37.94 ATOM 1652 C ASP 373 12.969 -5.833 24.577 1.00 15.09 ATOM 1653 O ASP 373 13.040 -4.928 25.407 1.00 37.94 ATOM 1654 N LEU 374 12.008 -5.901 23.659 1.00 17.04 ATOM 1655 CA LEU 374 10.974 -4.880 23.549 1.00 17.04 ATOM 1656 CB LEU 374 10.071 -5.15522.344 1.00 20.58 ATOM 1657 CG LEU 374 10.624 -4.720 20.985 1.00 20.58 ATOM 1658 CD1 LEU 374 9.826 -5.352 19.862 1.00 20.58 ATOM 1659 CD2 LEU 374 10.599 -3.202 20.882 1.00 20.58 ATOM 1660 C LEU 374 10.145 -4.786 24.825 1.00 17.04 ATOM 1661 OLEU 374 9.783 -3.688 25.256 1.00 20.58 ATOM 1662 N ARG 375 9.850 -5.935 25.430 1.00 20.46 ATOM 1663 CA ARG 375 9.080 -5.977 26.673 1.00 20.46 ATOM 1664 CB ARG 375 8.873 -7.422 27.140 1.00 55.89 ATOM 1665 CG ARG 375 8.180 -8.354 26.152 1.00 55.89 ATOM 1666 CD ARG 375 6.692 -8.084 26.027 1.00 55.89 ATOM 1667 NE ARG 375 5.943 -9.338 25.968 1.00 55.89 ATOM 1668 CZ ARG 375 5.054 -9.654 25.028 1.00 55.89 ATOM 1669 NH1 ARG 375 4.782 -8.808 24.040 1.00 55.89 ATOM 1670 NH2 ARG 375 4.438 -10.82925.073 1.00 55.89 ATOM 1671 C ARG 375 9.874 -5.221 27.735 1.00 20.46 ATOM 1672 O ARG 375 9.328 -4.391 28.463 1.00 55.89 ATOM 1673 N MET 376 11.174 -5.502 27.794 1.00 20.10 ATOM 1674 CA MET 376 12.076 -4.863 28.744 1.00 20.10 ATOM 1675 CB MET376 13.493 -5.417 28.580 1.00 63.73 ATOM 1676 CG MET 376 13.956 -6.310 29.722 1.00 63.73 ATOM 1677 SD MET 376 14.494 -5.373 31.182 1.00 63.73 ATOM 1678 CE MET 376 12.934 -5.151 32.087 1.00 63.73 ATOM 1679 C MET 376 12.081 -3.347 28.566 1.00 20.10 ATOM 1680 O MET 376 11.973 -2.602 29.539 1.00 63.73 ATOM 1681 N ILE 377 12.194 -2.896 27.321 1.00 30.02 ATOM 1682 CA ILE 377 12.198 -1.469 27.014 1.00 30.02 ATOM 1683 CB ILE 377 12.329 -1.228 25.488 1.00 19.31 ATOM 1684 CG2 ILE 377 12.0880.242 25.152 1.00 19.31 ATOM 1685 CG1 ILE 377 13.711 -1.685 25.011 1.00 19.31 ATOM 1686 CD1 ILE 377 13.906 -1.634 23.507 1.00 19.31 ATOM 1687 C ILE 377 10.915 -0.821 27.542 1.00 30.02 ATOM 1688 O ILE 377 10.962 0.216 28.211 1.00 19.31 ATOM 1689N GLY 378 9.779 -1.455 27.266 1.00 21.85 ATOM 1690 CA GLY 378 8.505 -0.936 27.729 1.00 21.85 ATOM 1691 C GLY 378 8.459 -0.821 29.243 1.00 21.85 ATOM 1692 O GLY 378 7.990 0.185 29.779 1.00 34.01 ATOM 1693 N ALA 379 8.967 -1.842 29.928 1.00 31.30 ATOM 1694 CA ALA 379 8.996 -1.870 31.388 1.00 31.30 ATOM 1695 CB ALA 379 9.471 -3.231 31.880 1.00 30.06 ATOM 1696 C ALA 379 9.895 -0.763 31.938 1.00 31.30 ATOM 1697 O ALA 379 9.482 0.002 32.810 1.00 30.06 ATOM 1698 N CYS 380 11.117 -0.677 31.4181.00 28.61 ATOM 1699 CA CYS 380 12.067 0.349 31.841 1.00 28.61 ATOM 1700 CB CYS 380 13.360 0.268 31.025 1.00 60.26 ATOM 1701 SG CYS 380 14.499 -1.067 31.470 1.00 60.26 ATOM 1702 C CYS 380 11.449 1.730 31.658 1.00 28.61 ATOM 1703 O CYS 38011.516 2.573 32.554 1.00 60.26 ATOM 1704 N HIS 381 10.840 1.957 30.498 1.00 30.42 ATOM 1705 CA HIS 381 10.212 3.243 30.216 1.00 30.42 ATOM 1706 CB HIS 381 9.696 3.306 28.779 1.00 16.49 ATOM 1707 CG HIS 381 8.942 4.562 28.472 1.00 16.49 ATOM1708 CD2 HIS 381 9.370 5.805 28.151 1.00 16.49 ATOM 1709 ND1 HIS 381 7.566 4.633 28.524 1.00 16.49 ATOM 1710 CE1 HIS 381 7.180 5.866 28.251 1.00 16.49 ATOM 1711 NE2 HIS 381 8.255 6.596 28.021 1.00 16.49 ATOM 1712 C HIS 381 9.073 3.539 31.182 1.00 30.42 ATOM 1713 O HIS 381 8.856 4.690 31.552 1.00 16.49 ATOM 1714 N ALA 382 8.330 2.506 31.564 1.00 22.89 ATOM 1715 CA ALA 382 7.218 2.666 32.493 1.00 22.89 ATOM 1716 CB ALA 382 6.520 1.336 32.708 1.00 34.50 ATOM 1717 C ALA 382 7.738 3.21333.819 1.00 22.89 ATOM 1718 O ALA 382 7.219 4.200 34.343 1.00 34.50 ATOM 1719 N SER 383 8.789 2.586 34.336 1.00 26.39 ATOM 1720 CA SER 383 9.400 3.006 35.591 1.00 26.39 ATOM 1721 CB SER 383 10.510 2.030 35.985 1.00 52.94 ATOM 1722 OG SER 38310.015 0.702 36.046 1.00 52.94 ATOM 1723 C SER 383 9.966 4.418 35.470 1.00 26.39 ATOM 1724 O SER 383 9.772 5.253 36.357 1.00 52.94 ATOM 1725 N ARG 384 10.662 4.683 34.368 1.00 30.36 ATOM 1726 CA ARG 384 11.249 5.995 34.134 1.00 30.36 ATOM 1727CB ARG 384 12.116 5.977 32.874 1.00 37.39 ATOM 1728 CG ARG 384 12.601 7.344 32.431 1.00 37.39 ATOM 1729 CD ARG 384 14.070 7.321 32.060 1.00 37.39 ATOM 1730 NE ARG 384 14.935 7.597 33.204 1.00 37.39 ATOM 1731 CZ ARG 384 15.750 8.646 33.291 1.00 37.39 ATOM 1732 NH1 ARG 384 15.824 9.529 32.303 1.00 37.39 ATOM 1733 NH2 ARG 384 16.488 8.819 34.376 1.00 37.39 ATOM 1734 C ARG 384 10.169 7.067 34.030 1.00 30.36 ATOM 1735 O ARG 384 10.301 8.144 34.616 1.00 37.39 ATOM 1736 N PHE 385 9.078 6.74933.338 1.00

24.47 ATOM 1737 CA PHE 385 7.980 7.693 33.171 1.00 24.47 ATOM 1738 CB PHE 385 6.859 7.092 32.319 1.00 28.70 ATOM 1739 CG PHE 385 5.710 8.036 32.075 1.00 28.70 ATOM 1740 CD1 PHE 385 5.795 9.017 31.092 1.00 28.70 ATOM 1741 CD2 PHE 3854.549 7.954 32.836 1.00 28.70 ATOM 1742 CE1 PHE 385 4.740 9.903 30.874 1.00 28.70 ATOM 1743 CE2 PHE 385 3.491 8.835 32.624 1.00 28.70 ATOM 1744 CZ PHE 385 3.587 9.812 31.641 1.00 28.70 ATOM 1745 C PHE 385 7.436 8.097 34.533 1.00 24.47 ATOM 1746O PHE 385 7.250 9.285 34.805 1.00 28.70 ATOM 1747 N LEU 386 7.208 7.107 35.391 1.00 31.13 ATOM 1748 CA LEU 386 6.690 7.352 36.734 1.00 31.13 ATOM 1749 CB LEU 386 6.596 6.044 37.513 1.00 39.10 ATOM 1750 C LEU 386 7.577 8.348 37.474 1.00 31.13 ATOM 1751 O LEU 386 7.085 9.201 38.217 1.00 39.10 ATOM 1752 N HIS 387 8.884 8.254 37.243 1.00 36.46 ATOM 1753 CA HIS 387 9.837 9.152 37.881 1.00 36.46 ATOM 1754 CB HIS 387 11.258 8.589 37.794 1.00 62.78 ATOM 1755 CG HIS 387 11.459 7.338 38.5901.00 62.78 ATOM 1756 CD2 HIS 387 10.601 6.614 39.346 1.00 62.78 ATOM 1757 ND1 HIS 387 12.675 6.689 38.663 1.00 62.78 ATOM 1758 CE1 HIS 387 12.554 5.620 39.431 1.00 62.78 ATOM 1759 NE2 HIS 387 11.309 5.550 39.856 1.00 62.78 ATOM 1760 C HIS 3879.778 10.544 37.266 1.00 36.46 ATOM 1761 O HIS 387 9.885 11.543 37.979 1.00 62.78 ATOM 1762 N MET 388 9.587 10.612 35.950 1.00 33.41 ATOM 1763 CA MET 388 9.505 11.894 35.258 1.00 33.41 ATOM 1764 CB MET 388 9.269 11.703 33.755 1.00 42.63 ATOM1765 CG MET 388 10.456 11.144 32.982 1.00 42.63 ATOM 1766 SD MET 388 10.253 11.325 31.192 1.00 42.63 ATOM 1767 CE MET 388 9.501 9.772 30.748 1.00 42.63 ATOM 1768 C MET 388 8.385 12.746 35.849 1.00 33.41 ATOM 1769 O MET 388 8.573 13.934 36.1031.00 42.63 ATOM 1770 N LYS 389 7.235 12.126 36.092 1.00 39.26 ATOM 1771 CA LYS 389 6.082 12.825 36.659 1.00 39.26 ATOM 1772 CB LYS 389 4.867 11.900 36.719 1.00 52.87 ATOM 1773 CG LYS 389 4.237 11.594 35.379 1.00 52.87 ATOM 1774 CD LYS 389 3.04810.667 35.553 1.00 52.87 ATOM 1775 CE LYS 389 3.482 9.321 36.125 1.00 52.87 ATOM 1776 NZ LYS 389 2.335 8.407 36.326 1.00 52.87 ATOM 1777 C LYS 389 6.363 13.360 38.056 1.00 39.26 ATOM 1778 O LYS 389 5.837 14.404 38.452 1.00 52.87 ATOM 1779 N VAL390 7.156 12.614 38.818 1.00 44.18 ATOM 1780 CA VAL 390 7.508 13.016 40.172 1.00 44.18 ATOM 1781 CB VAL 390 8.299 11.898 40.905 1.00 50.50 ATOM 1782 CG1 VAL 390 8.718 12.362 42.293 1.00 50.50 ATOM 1783 CG2 VAL 390 7.455 10.640 41.012 1.00 50.50 ATOM 1784 C VAL 390 8.352 14.288 40.145 1.00 44.18 ATOM 1785 O VAL 390 8.144 15.198 40.948 1.00 50.50 ATOM 1786 N GLU 391 9.261 14.368 39.179 1.00 38.64 ATOM 1787 CA GLU 391 10.161 15.509 39.056 1.00 38.64 ATOM 1788 CB GLU 391 11.483 15.06038.424 1.00 64.18 ATOM 1789 CG GLU 391 12.065 13.766 39.009 1.00 64.18 ATOM 1790 CD GLU 391 12.662 13.922 40.405 1.00 64.18 ATOM 1791 OE1 GLU 391 12.190 14.773 41.192 1.00 64.18 ATOM 1792 OE2 GLU 391 13.611 13.173 40.721 1.00 64.18 ATOM 1793 CGLU 391 9.623 16.737 38.314 1.00 38.64 ATOM 1794 O GLU 391 9.656 17.850 38.849 1.00 64.18 ATOM 1795 N CYS 392 9.125 16.539 37.096 1.00 37.24 ATOM 1796 CA CYS 392 8.611 17.635 36.271 1.00 37.24 ATOM 1797 CB CYS 392 8.879 17.345 34.784 1.00 30.64 ATOM 1798 SG CYS 392 10.634 17.137 34.283 1.00 30.64 ATOM 1799 C CYS 392 7.110 17.882 36.496 1.00 37.24 ATOM 1800 O CYS 392 6.403 17.011 37.006 1.00 30.64 ATOM 1801 N PRO 393 6.625 19.107 36.199 1.00 40.56 ATOM 1802 CD PRO 393 7.444 20.297 35.9041.00 33.41 ATOM 1803 CA PRO 393 5.209 19.473 36.358 1.00 40.56 ATOM 1804 CB PRO 393 5.253 21.001 36.404 1.00 33.41 ATOM 1805 CG PRO 393 6.409 21.332 35.527 1.00 33.41 ATOM 1806 C PRO 393 4.330 18.975 35.207 1.00 40.56 ATOM 1807 O PRO 393 4.77618.907 34.057 1.00 33.41 ATOM 1808 N THR 394 3.067 18.691 35.516 1.00 41.91 ATOM 1809 CA THR 394 2.101 18.186 34.540 1.00 41.91 ATOM 1810 CB THR 394 0.691 18.075 35.156 1.00 62.04 ATOM 1811 OG1 THR 394 0.706 18.582 36.497 1.00 62.04 ATOM 1812CG2 THR 394 0.232 16.626 35.168 1.00 62.04 ATOM 1813 C THR 394 1.995 18.984 33.242 1.00 41.91 ATOM 1814 O THR 394 1.758 18.411 32.181 1.00 62.04 ATOM 1815 N GLU 395 2.191 20.297 33.327 1.00 43.92 ATOM 1816 CA GLU 395 2.104 21.176 32.160 1.00 43.92 ATOM 1817 CB GLU 395 2.313 22.626 32.585 1.00 34.22 ATOM 1818 C GLU 395 3.071 20.814 31.031 1.00 43.92 ATOM 1819 O GLU 395 2.887 21.243 29.891 1.00 34.22 ATOM 1820 N LEU 396 4.104 20.041 31.350 1.00 34.92 ATOM 1821 CA LEU 396 5.096 19.63430.359 1.00 34.92 ATOM 1822 CB LEU 396 6.473 19.495 31.017 1.00 35.81 ATOM 1823 CG LEU 396 7.074 20.747 31.662 1.00 35.81 ATOM 1824 CD1 LEU 396 8.427 20.410 32.263 1.00 35.81 ATOM 1825 CD2 LEU 396 7.209 21.857 30.629 1.00 35.81 ATOM 1826 C LEU396 4.731 18.324 29.661 1.00 34.92 ATOM 1827 O LEU 396 5.343 17.954 28.659 1.00 35.81 ATOM 1828 N PHE 397 3.734 17.627 30.197 1.00 35.28 ATOM 1829 CA PHE 397 3.302 16.352 29.640 1.00 35.28 ATOM 1830 CB PHE 397 3.059 15.341 30.764 1.00 27.13 ATOM 1831 CG PHE 397 4.285 15.004 31.561 1.00 27.13 ATOM 1832 CD1 PHE 397 4.700 15.824 32.604 1.00 27.13 ATOM 1833 CD2 PHE 397 5.021 13.860 31.273 1.00 27.13 ATOM 1834 CE1 PHE 397 5.831 15.510 33.349 1.00 27.13 ATOM 1835 CE2 PHE 397 6.155 13.53732.013 1.00 27.13 ATOM 1836 CZ PHE 397 6.561 14.364 33.052 1.00 27.13 ATOM 1837 C PHE 397 2.027 16.474 28.812 1.00 35.28 ATOM 1838 O PHE 397 0.977 16.861 29.331 1.00 27.13 ATOM 1839 N PRO 398 2.102 16.164 27.505 1.00 26.41 ATOM 1840 CD PRO 3983.305 15.850 26.713 1.00 19.32 ATOM 1841 CA PRO 398 0.917 16.247 26.647 1.00 26.41 ATOM 1842 CB PRO 398 1.439 15.752 25.300 1.00 19.32 ATOM 1843 CG PRO 398 2.867 16.193 25.312 1.00 19.32 ATOM 1844 C PRO 398 -0.157 15.313 27.206 1.00 26.41 ATOM1845 O PRO 398 0.160 14.232 27.710 1.00 19.32 ATOM 1846 N PRO 399 -1.439 15.702 27.104 1.00 25.12 ATOM 1847 CD PRO 399 -1.935 16.929 26.454 1.00 24.32 ATOM 1848 CA PRO 399 -2.554 14.894 27.612 1.00 25.12 ATOM 1849 CB PRO 399 -3.777 15.594 27.0221.00 24.32 ATOM 1850 CG PRO 399 -3.349 17.026 26.974 1.00 24.32 ATOM 1851 C PRO 399 -2.502 13.416 27.222 1.00 25.12 ATOM 1852 O PRO 399 -2.599 12.540 28.085 1.00 24.32 ATOM 1853 N LEU 400 -2.322 13.139 25.933 1.00 23.10 ATOM 1854 CA LEU 400-2.265 11.759 25.454 1.00 23.10 ATOM 1855 CB LEU 400 -2.230 11.720 23.923 1.00 22.35 ATOM 1856 CG LEU 400 -2.485 10.354 23.276 1.00 22.35 ATOM 1857 CD1 LEU 400 -3.792 9.765 23.792 1.00 22.35 ATOM 1858 CD2 LEU 400 -2.523 10.494 21.763 1.00 22.35 ATOM 1859 C LEU 400 -1.066 11.012 26.032 1.00 23.10 ATOM 1860 O LEU 400 -1.160 9.825 26.345 1.00 22.35 ATOM 1861 N PHE 401 0.044 11.723 26.202 1.00 13.85

ATOM 1862 CA PHE 401 1.269 11.150 26.755 1.00 13.85 ATOM 1863 CB PHE 401 2.374 12.213 26.753 1.00 26.97 ATOM 1864 CG PHE 401 3.729 11.702 27.164 1.00 26.97 ATOM 1865 CD1 PHE 401 4.189 10.461 26.732 1.00 26.97 ATOM 1866 CD2 PHE 401 4.56112.481 27.963 1.00 26.97 ATOM 1867 CE1 PHE 401 5.459 10.005 27.091 1.00 26.97 ATOM 1868 CE2 PHE 401 5.830 12.035 28.327 1.00 26.97 ATOM 1869 CZ PHE 401 6.280 10.795 27.889 1.00 26.97 ATOM 1870 C PHE 401 0.993 10.659 28.179 1.00 13.85 ATOM 1871O PHE 401 1.393 9.558 28.555 1.00 26.97 ATOM 1872 N LEU 402 0.274 11.473 28.947 1.00 25.21 ATOM 1873 CA LEU 402 -0.080 11.145 30.325 1.00 25.21 ATOM 1874 CB LEU 402 -0.640 12.380 31.035 1.00 29.34 ATOM 1875 CG LEU 402 0.334 13.411 31.600 1.00 29.34 ATOM 1876 CD1 LEU 402 -0.430 14.658 32.018 1.00 29.34 ATOM 1877 CD2 LEU 402 1.090 12.814 32.775 1.00 29.34 ATOM 1878 C LEU 402 -1.109 10.025 30.425 1.00 25.21 ATOM 1879 O LEU 402 -1.034 9.189 31.320 1.00 29.34 ATOM 1880 N GLU 403 -2.09010.043 29.529 1.00 23.54 ATOM 1881 CA GLU 403 -3.159 9.046 29.521 1.00 23.54 ATOM 1882 CB GLU 403 -4.274 9.482 28.562 1.00 63.22 ATOM 1883 CG GLU 403 -5.469 8.531 28.506 1.00 63.22 ATOM 1884 CD GLU 403 -6.530 8.952 27.498 1.00 63.22 ATOM 1885OE1 GLU 403 -6.237 9.786 26.613 1.00 63.22 ATOM 1886 OE2 GLU 403 -7.666 8.436 27.589 1.00 63.22 ATOM 1887 C GLU 403 -2.708 7.629 29.170 1.00 23.54 ATOM 1888 O GLU 403 -3.210 6.656 29.735 1.00 63.22 ATOM 1889 N VAL 404 -1.787 7.515 28.221 1.00 33.24 ATOM 1890 CA VAL 404 -1.297 6.213 27.782 1.00 33.24 ATOM 1891 CB VAL 404 -0.621 6.314 26.390 1.00 30.71 ATOM 1892 CG1 VAL 404 -0.097 4.957 25.947 1.00 30.71 ATOM 1893 CG2 VAL 404 -1.611 6.841 25.371 1.00 30.71 ATOM 1894 C VAL 404 -0.3385.528 28.752 1.00 33.24 ATOM 1895 O VAL 404 -0.386 4.305 28.914 1.00 30.71 ATOM 1896 N PHE 405 0.526 6.309 29.392 1.00 33.66 ATOM 1897 CA PHE 405 1.516 5.752 30.308 1.00 33.66 ATOM 1898 CB PHE 405 2.901 6.326 29.984 1.00 34.35 ATOM 1899 CG PHE405 3.343 6.076 28.568 1.00 34.35 ATOM 1900 CD1 PHE 405 3.519 7.134 27.683 1.00 34.35 ATOM 1901 CD2 PHE 405 3.569 4.782 28.114 1.00 34.35 ATOM 1902 CE1 PHE 405 3.911 6.906 26.365 1.00 34.35 ATOM 1903 CE2 PHE 405 3.960 4.545 26.798 1.00 34.35 ATOM 1904 CZ PHE 405 4.131 5.610 25.922 1.00 34.35 ATOM 1905 C PHE 405 1.189 5.931 31.790 1.00 33.66 ATOM 1906 O PHE 405 2.036 5.539 32.623 1.00 34.35 ATOM 1907 OXT PHE 405 0.090 6.434 32.107 1.00 34.35 ATOM 1908 C1 TRI 1 8.375 7.063 18.475 1.00 34.21 ATOM 1909 C2 TRI 1 10.048 8.688 23.016 1.00 33.36 ATOM 1910 C3 TRI 1 8.104 8.391 18.941 1.00 34.21 ATOM 1911 C4 TRI 1 10.496 9.696 23.813 1.00 33.36 ATOM 1912 C5 TRI 1 8.916 8.943 19.927 1.00 34.21 ATOM 1913 C6 TRI 1 10.152 9.772 25.1211.00 33.36 ATOM 1914 C7 TRI 1 9.862 8.178 20.609 1.00 34.21 ATOM 1915 C8 TRI 1 9.246 8.821 25.653 1.00 33.36 ATOM 1916 C9 TRI 1 10.117 6.865 20.147 1.00 34.21 ATOM 1917 C10 TRI 1 8.805 7.754 24.847 1.00 33.36 ATOM 1918 C11 TRI 1 9.375 6.33919.026 1.00 34.21 ATOM 1919 C12 TRI 1 9.125 7.756 23.490 1.00 33.36 ATOM 1920 C13 TRI 1 7.540 6.470 17.383 1.00 35.85 ATOM 1921 C15 TRI 1 8.158 6.555 15.938 1.00 35.85 ATOM 1922 I1 TRI 1 8.713 10.990 20.395 1.00 34.21 ATOM 1923 I2 TRI 1 10.95111.289 26.315 1.00 33.36 ATOM 1924 I3 TRI 1 11.592 5.685 21.118 1.00 34.21 ATOM 1925 O3 TRI 1 9.407 6.654 15.852 1.00 35.85 ATOM 1926 O2 TRI 1 10.570 8.649 21.717 1.00 33.36 ATOM 1927 O1 TRI 1 8.798 8.969 26.979 1.00 33.36 ATOM 1928 O4 TRI 17.352 6.522 14.973 1.00 35.85 ATOM 1929 O1 HOH 501 9.189 2.098 11.091 1.00 33.36 ATOM 1930 O1 HOH 503 5.152 5.261 12.137 1.00 33.36 ATOM 1931 O1 HOH 504 3.970 5.057 16.390 1.00 33.36 ATOM 1932 O1 HOH 534 8.296 -0.941 8.998 1.00 33.36 ATOM 1933O1 HOH 538 4.845 14.369 13.635 1.00 33.36 ATOM 1934 O1 HOH 540 5.789 12.049 10.352 1.00 33.36 ATOM 1936 O1 HOH 555 5.721 2.525 28.939 1.00 33.36 ATOM 1937 O1 HOH 556 3.732 1.273 26.724 1.00 33.36 ATOM 1935 O1 HOH 600 8.767 4.847 8.517 1.00 33.36 ATOM 1938 AS1 CAD 701 1.863 1.579 0.837 1.00 37.00 ATOM 1939 C2 CAD 701 1.760 -0.100 0.335 1.00 33.36 ATOM 1940 C3 CAD 701 3.511 1.872 1.858 1.00 28.02 ATOM 1941 O4 CAD 701 1.785 2.506 -0.433 1.00 28.02 ATOM 1942 O5 CAD 701 0.592 2.019 1.654 1.00 28.02 ATOM 1943 AS AS 801 11.254 16.718 33.126 1.00 37.00 AS ATOM 1944 AS AS 802 16.338 -1.161 29.914 1.00 37.00 AS ATOM 1945 AS AS 803 -14.931 -11.763 25.324 1.00 37.00 AS END

APPENDIX 5 TR_IPBR2.PDB REMARK rTR_ipbr2 full length numbering REMARK REMARK Rfactor 0.214 Rfree 0.224 REMARK Resolution 15. 2.2 all reflections REMARK REMARK Three cacodylate-modified cysteines (CYA) REMARK Cya334, Cya380, Cya392 REMARK cacodylate modeled as single arsenic atom REMARK REMARK side chain of certain residues modeled as ALA due to poor density; REMARK however, residue name reflects true residue for clarity REMARK REMARK clone obtained from Murray et. al. REMARK deposited sequence confirmed, REMARK differing from that reported by Thompson et. al. REMARK in the following codons: REMARK 281 Thr--Ala REMARK 285 Lys--Glu REMARK identical to that reported by Mitsuhashi et. al. REMARK gb:RNTRAVI X07409 JRNL AUTH M. B. MURRAY, N. D. ZILZ, N. L. MCCREARY, M. J. MACDONALD JRNL AUTH 2 H. C. TOWLE JRNL TITL ISOLATION AND CHARACTERIZATION OF RAT CDNA CLONES FOR TWO JRNL TITL 2 DISTINCT THYROID HORMONE RECPTORS JRNL REF JBC V. 263 25 1988 JRNL AUTH C.C. THOMPSON, C. WEINBERGER, R. LEBO, R. M. EVANS JRNL TITL IDENTIFICATION OF A NOVEL THYROID HORMONE RECEPTOR EXPRESSED JRNL TITL 2 IN THE MAMMALIAN CENTRAL NERVOUS SYSTEM JRNL REF SCIENCE V. 237 1987 JRNL AUTH T. MITSUHASHI, G. TENNYSON, V. NIKODEM JRNL TITL NUCLEOTIDE SEQUENCE OF NOVEL CDNAS GENERATED BY ALTERNATIVE JRNL TITL 2 SPLICING OF A RAT THYROID HORMONE RECEPTOR GENE TRANSCRIPT JRNL REF NUC. ACIDS. RES. V. 16 12 1988 REMARK ATOM 1 CB ARG 157 68.481 10.663 6.906 1.00 57.50 ATOM 2 CGARG 157 69.793 10.213 7.512 1.00 59.93 ATOM 3 CD ARG 157 70.510 11.365 8.189 1.00 70.24 ATOM 4 NE ARG 157 71.661 10.906 8.961 1.00 77.62 ATOM 5 CZ ARG 157 11.599 10.492 10.224 1.00 78.75 ATOM 6 NH1 ARG 157 70.440 10.480 10.870 1.00 74.33 ATOM 7NH2 ARG 157 72.697 10.075 10.839 1.00 83.44 ATOM 8 C ARG 157 66.314 10.014 5.809 1.00 46.84 ATOM 9 O ARG 157 66.109 10.397 4.659 1.00 54.49 ATOM 10 N ARG 157 68.442 9.069 5.013 1.00 56.54 ATOM 11 CA ARG 157 67.704 9.537 6.222 1.00 52.92 ATOM 12N PRO 158 65.335 9.953 6.727 1.00 39.44 ATOM 13 CD PRO 158 65.503 9.448 8.099 1.00 41.72 ATOM 14 CA PRO 158 63.946 10.368 6.487 1.00 34.98 ATOM 15 CB PRO 158 63.282 10.172 7.854 1.00 34.92 ATOM 16 CG PRO 158 64.096 9.096 8.487 1.00 45.83 ATOM17 C PRO 158 63.765 11.804 5.992 1.00 34.13 ATOM 18 O PRO 158 64.223 12.757 6.621 1.00 31.07 ATOM 19 N GLU 159 63.110 11.932 4.841 1.00 31.36 ATOM 20 CA GLU 159 62.814 13.220 4.228 1.00 27.34 ATOM 21 CB GLU 159 62.569 13.041 2.726 1.00 24.27 ATOM 22 CG GLU 159 63.814 12.866 1.887 1.00 24.85 ATOM 23 CD GLU 159 64.409 14.188 1.454 1.00 28.12 ATOM 24 OE1 GLU 159 63.642 15.144 1.224 1.00 29.26 ATOM 25 OE2 GLU 159 65.646 14.269 1.326 1.00 29.52 ATOM 26 C GLU 159 61.528 13.707 4.870 1.00 24.30 ATOM 27 O GLU 159 60.855 12.934 5.566 1.00 29.01 ATOM 28 N PRO 160 61.192 14.989 4.718 1.00 24.62 ATOM 29 CD PRO 160 61.979 16.126 4.188 1.00 18.72 ATOM 30 CA PRO 160 59.947 15.451 5.330 1.00 21.62 ATOM 31 CB PRO 160 59.945 16.955 5.0481.00 12.71 ATOM 32 CG PRO 160 61.394 17.297 4.930 1.00 15.12 ATOM 33 C PRO 160 58.743 14.752 4.671 1.00 24.61 ATOM 34 O PRO 160 58.789 14.384 3.490 1.00 22.63 ATOM 35 N THR 161 57.705 14.504 5.450 1.00 25.86 ATOM 36 CA THR 161 56.515 13.8644.921 1.00 23.77 ATOM 37 CB THR 161 55.689 13.201 6.048 1.00 21.75 ATOM 38 OG1 THR 161 55.178 14.210 6.926 1.00 20.78 ATOM 39 CG2 THR 161 56.549 12.227 6.847 1.00 18.44 ATOM 40 C THR 161 55.680 14.967 4.269 1.00 28.67 ATOM 41 O THR 161 55.91716.151 4.510 1.00 29.90 ATOM 42 N PRO 162 54.685 14.597 3.448 1.00 27.79 ATOM 43 CD PRO 162 54.313 13.237 3.019 1.00 23.25 ATOM 44 CA PRO 162 53.843 15.603 2.795 1.00 26.19 ATOM 45 CB PRO 162 52.699 14.766 2.227 1.00 19.89 ATOM 46 CG PRO 16253.394 13.492 1.848 1.00 20.63 ATOM 47 C PRO 162 53.334 16.661 3.775 1.00 24.81 ATOM 48 O PRO 162 53.477 17.863 3.526 1.00 21.10 ATOM 49 N GLU 163 52.812 16.198 4.911 1.00 26.34 ATOM 50 CA GLU 163 52.266 17.065 5.959 1.00 30.38 ATOM 51 CB GLU163 51.640 16.231 7.086 1.00 29.46 ATOM 52 CG GLU 163 50.482 15.321 6.666 1.00 48.37 ATOM 53 CD GLU 163 50.918 14.132 5.816 1.00 53.12 ATOM 54 OE1 GLU 163 51.890 13.441 6.194 1.00 52.22 ATOM 55 OE2 GLU 163 50.282 13.886 4.766 1.00 59.14 ATOM 56C GLU 163 53.353 17.949 6.552 1.00 26.74 ATOM 57 O GLU 163 53.109 19.107 6.898 1.00 27.03 ATOM 58 N GLU 164 54.553 17.389 6.677 1.00 26.74 ATOM 59 CA GLU 164 55.679 18.124 7.221 1.00 23.65 ATOM 60 CB GLU 164 56.805 17.174 7.609 1.00 18.85 ATOM61 CG GLU 164 56.441 16.306 8.804 1.00 26.81 ATOM 62 CD GLU 164 57.536 15.334 9.188 1.00 31.06 ATOM 63 OE1 GLU 164 58.404 15.050 8.340 1.00 29.21 ATOM 64 OE2 GLU 164 57.524 14.848 10.340 1.00 31.39 ATOM 65 C GLU 164 56.165 19.204 6.276 1.00 26.54 ATOM 66 O GLU 164 56.609 20.258 6.724 1.00 32.48 ATOM 67 N TRP 165 56.075 18.957 4.971 1.00 23.41 ATOM 68 CA TRP 165 56.488 19.962 3.998 1.00 20.81 ATOM 69 CB TRP 165 56.462 19.405 2.573 1.00 18.15 ATOM 70 CG TRP 165 57.762 18.747 2.1641.00 15.80 ATOM 71 CD2 TRP 165 59.058 19.377 2.064 1.00 15.35 ATOM 72 CE2 TRP 165 59.959 18.392 1.628 1.00 12.14 ATOM 73 CE3 TRP 165 59.527 20.676 2.287 1.00 17.56 ATOM 74 CD1 TRP 165 57.939 17.449 1.804 1.00 12.78 ATOM 75 NE1 TRP 165 59.25317.230 1.484 1.00 16.10 ATOM 76 CZ2 TRP 165 61.318 18.657 1.419 1.00 16.26 ATOM 77 CZ3 TRP 165 60.879 20.944 2.079 1.00 19.52 ATOM 78 CH2 TRP 165 61.760 19.933 1.642 1.00 16.48 ATOM 79 C TRP 165 55.547 21.151 4.109 1.00 19.66 ATOM 80 O TRP 16555.975 22.295 3.960 1.00 23.61 ATOM 81 N ASP 166 54.269 20.882 4.376 1.00 22.66 ATOM 82 CA ASP 166 53.269 21.943 4.537 1.00 23.35 ATOM 83 CB ASP 166 51.863 21.359 4.716 1.00 22.61 ATOM 84 CG ASP 166 51.347 20.681 3.458 1.00 31.41 ATOM 85 OD1ASP 166 51.816 21.028 2.360 1.00 26.38 ATOM 86 OD2 ASP 166 50.464 19.803 3.570 1.00 32.25 ATOM 87 C ASP 166 53.631 22.760 5.773 1.00 26.47 ATOM 88 O ASP 166 53.694 23.991 5.718 1.00 30.25 ATOM 89 N LEU 167 53.887 22.054 6.872 1.00 24.12 ATOM 90CA LEU 167 54.268 22.663 8.139 1.00 26.44 ATOM 91 CB LEU 167 54.596 21.557 9.148 1.00 32.57 ATOM 92 CG LEU 167 54.659 21.919 10.629 1.00 36.97 ATOM 93 CD1 LEU 167 53.289 22.402 11.080 1.00 43.83 ATOM 94 CD2 LEU 167 55.096 20.712 11.448 1.00 34.75 ATOM 95 C LEU 167 55.501 23.533 7.904 1.00 23.19 ATOM 96 O LEU 167 55.570 24.670 8.368 1.00 28.18 ATOM 97 N ILE 168 56.450 22.988 7.147 1.00 19.25 ATOM 98 CA ILE 168 57.703 23.651 6.801 1.00 17.71 ATOM 99 CB ILE 168 58.632 22.693 6.0061.00 14.43 ATOM 100 CG2 ILE 168 59.740 23.451 5.304 1.00 16.71 ATOM 101 CG1 ILE 168 59.219 21.644 6.948 1.00 21.24 ATOM 102 CD1 ILE 168 60.063 20.588 6.264 1.00 18.18 ATOM 103 C ILE 168 57.475 24.931 6.002 1.00 28.73 ATOM 104 O ILE 168 58.06425.977 6.307 1.00 29.36 ATOM 105 N HIS 169 56.601 24.866 5.005 1.00

24.43 ATOM 106 CA HIS 169 56.319 26.027 4.169 1.00 23.64 ATOM 107 CB HIS 169 55.459 25.631 2.971 1.00 23.55 ATOM 108 CG HIS 169 56.140 24.683 2.034 1.00 23.82 ATOM 109 CD2 HIS 169 57.455 24.429 1.824 1.00 19.23 ATOM 110 ND1 HIS 16955.450 23.833 1.199 1.00 22.9