AU9611398A - Crystal structure and mutants of interleukin-1beta converting enzyme - Google Patents

Description

I- 1

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT r ~r ro o c rr Name of Applicant: VERTEX

INCORPORATED

PHARMACEUTICALS

Actual inventors: Keith P WILSON, James P GRIFFITH, Eunice E KIM, David J LIVINGSTON Address for Service: CULLEN CO., Patent Trade Mark Attorneys, 240 Queen Street, Brisbane, Qld. 4000, Australia.

Invention Title: CRYSTAL STRUCTURE AND MUTANTS OF INTERLEUKIN-1 CONVERTING

ENZYME

p- 7,.

Divisional Application of 27055/95 filed 16.6.95 The following statement is a full description of this invention, including the best method of performing it known to us Rkp.

i~L I" rUI~-- l lr~L4P rr~ ~mOW II-MI I CRYSTAL STRUCTURE AND MUTANTS OF INTERLEUKIN-19 CONVERTING ENZYME TECHNICAL FIELD OF INVENTION The present invention relates to crystals of interleukin-lZ converting enzyme and more particularly to the high resolution structure of ICE S".obtained by X-ray diffraction. This invention also relates to mutants of ICE. In addition, this invention relates to methods of using the structure coordinates of ICE and mutants thereof to screen and design compounds that bind to the active site and accessory S"binding site of ICE.

BACKGROUND ART Interleukin-1 is a major pro- S 15 inflammatory and immunoregulatory protein that stimulates fibroblast differentiation and proliferation, the production of prostaglandins, collagenase and phospholipase by synovial cells and chondrocytes, bas6phil and eosinophil degranulation and neutrophil activation. Oppenheim, J.H. et al, Immunology Today, 7, pp. 45-56 (1986). As such, it is involved in the pathogenesis of chronic and acute inflammatory and autoimmune diseases. IL-1 is predominantly produced by peripheral blood monocytes i25 and exists in two distinct agonist forms, IL-1t and IL- ~1 I. Mosely, B.S. et al., Proc. Nat. Acad. Sci., 84, A f pp. 4572-4576 Lonnemann G. et al., Eur.j.

Immunol., 19, pp. 1531-1536 (1989).

IL-1i9 is synthesized as a biologically inactive precursor, pIL-ig. pIL-f? is a 33kDa polypeptide that lacks a conventional leader sequence and is not processed by a signal peptidase. March, Nature, 315, pp. 641-647 (1985). Instead, pIL-19 is cleaved by interleukin-19 converting enzyme (RICE11) between Asp 116 and Ala 117 to produce the biologically active C-terminal fragmnent of 17kDa molecular weight found in serum and synovial fluid. Sleath, P.R.

et al., J. Biol. Chem., 265, pp. 14526-14528 (1992); Howard, A.D. et al., J. Immunol., 147, pp. 2964-2969 (1991) Processing by ICE is also necessary for the transport of mature IL-19 through the cell membrane.

ICE is a cysteine protease localized primarily in monocytes. It converts precursor IL-1f? to the mature form. Black, R.A. et al., FEES Lett., 247, .so pp. 386-390 (1989); Kostura, M.J. et al., Proc. Natl.

20 Acad. Sci. USA, 86, pp. 5227-5231. (1989) ICE, or its homologues, also appears to be involved in the regulation of cell death or apoptosis. Yuan, J.

et al., Cell, 75, pp. 641-652 (1993); Miura, M. et al., Cell, 75, pp. 653-660 (1993); Nett-Fiordalisi, M.A.

~25 et al., J. Cell Biochem., 17B, p. 117 (1993). In particular, ICE or ICE homologues are thought to be associated with the regulation of apoptosis in neurogenerative diseases, such as Alzheimer's and Parkinson's disease. Marx, J. and M. Baringa, Science, 259, pp. 760-762 (1993); Gagliardini, V. et al., Science, 263, pp. 826-828 (1994).

ICE has been previously described as a heterodimer composed of two subunits, p20 and (2OkDa and l0kDa molecular weight, respectively).

These subunits are derived from a 45kDa proenzyme (p4 -3by way of a p30 form, through an activation mechanism that is autocatalytic. Thornberry, N.A. et al., Nature, 356, pp. 768-774 (1992). The ICE proenzyme has been divided into several functional domains: a prodomain (p14), a p22/ 20 subunit, a polypeptide linker and a plO subunit. Thornberry et al., suDra; Casano et al., Genomics, 20, pp. 474-481 (1994).

Full length p45 has been characterized by its cDNA and amino acid sequences. PCT patent applications S 10 WO91/15577 and WO 94/00154. The p 20 and p10 cDNA and amino acid sequences are also known. Thornberry et al., sura. Murine and rat ICE have also been sequenced and cloned. They have high amino acid and nucleic acid sequence homology to human ICE. Miller, 15 D.K. et al., Ann. N.Y. Acad. Sci., 696, pp. 133-148 (1993); Molineaux, S.M. et al., Proc. Nat. Acad. Sci., pp. 1809-1813 (1993). Knowledge of the primary structure of ICE, however, does not allow prediction of its tertiary structure. Nor does it afford an 20 understanding of the structural, conformational and chemical interactions of ICE and its substrate pIL-l9 or other substrates or inhibitors.

ICE inhibitors represent a class of compounds S"useful for the control of inflammation or apoptosis or 25 both. Peptide and peptidyl inhibitors of ICE have been described. PCT patent applications WO 91/15577;

WO

93/05071; WO 93/09135; WO 93/14777 and WO 93/16710; and European patent application 0 547 699. However, due to their peptidic nature, such inhibitors are typically characterized by undesirable pharmacologic properties, such as poor oral absorption, poor stability and rapid metabolism. Plattner, J.J. and D.W. Norbeck, in Drug Discovery Technologies, C.R. Clark and W.H. Moos, Eds.

(Ellis Horwood, Chichester, England, 1990), pp. 92i r4 ~1~11~1 i -4- 126. This has hampered their development into effective drugs.

SUMMARY OF THE INVENTION The present invention solves the above problems.

It is an object of this invention to solve the three-dimensional structure of interleukin-1i converting enzyme and to determine its structure coordinates.

S 10 It is an object of this invention to use the structure coordinates of an ICE crystal to reveal the atomic details of the active site and one or more accessory binding sites of the enzyme.

It is also an object of this invention to use the structure coordinates of an ICE crystal to solve the structure of a different ICE crystal, or a crystal of a mutant, homologue or co-complex, of ICE.

It is a further object of this invention to provide interleukin-l1 converting enzyme mutants characterized by one or more different properties as compared with wild-type ICE. These properties include altered surface charge, increased stability to subunit Sdissociation, altered substrate specificity or higher specific activity. ICE mutants are useful to identify those amino acids that are most important for the Senzymatic activity of ICE. This information, in turn, allows the design of improved inhibitors of ICE as compared with peptidic ICE inhibitors.

It is also an object of this invention to use the structure coordinates and atomic details of ICE, or its mutants or homologues or co-complexes, to design, evaluate computationally, synthesize and use inhibitors of ICE that avoid the undesirable physical and pharmacologic properties of peptidic ICE inhibitors.

7Viii birr~-;r~-Prrmr~F~man~~ 1 BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 represents a ribbon drawing of the p20/pl0 interleukin-l converting enzyme heterodimer.

The active site is at the top of the figure, roughly at the center of the cluster of displayed side chains.

Figure 2 represents a space-filling model of the (p20) 2 /(pl0) 2 tetramer of interleukin-lI converting enzyme. Two p20 subunits (dark shade) are in contact with two p10 subunits (light shade). Black shading on 10 top left and bottom right represents a tetrapeptide aldehyde inhibitor bound in each of the two active sites of the tetramer. The crystallographic two-fold axis is approximately perpendicular to the plane of drawing, and runs through the small hole at the center of the interface between the two pl0 subunits. The Nand C-terminal ends qf each subunit are labeled.

Figure 3 is a graphic depiction of the S activity of various interleukin-lf converting enzyme mutants in processing pIL-19 intracellularly, relative 20 to wild-type interleukin-19 converting enzyme activity.

S, The particular mutants tested are designated on the x-axis using nomenclature listing the specific amino acid and its residue number. For example, indicates replacement of amino acid Cys-285 with serine. Activity levels were measured at 16 hours (hatched bar) and 24 hours (solid bars).

BRIEF DESCRIPTION OF THE TABLES Table A lists the amino acids of ICE that constitute the tetramer interface contacts between the ICE subunits and that constitute the accessory binding site moiety.

j Table B lists the atomic structure g -coordinates for ICE as derived by X-ray diffraction from a crystal of ICE complexed to a tetrapeptide inhibitor.

ABBREVIATIONS AND DEFINITIONS ABBr~EVIATIONS Amino Acids A =Ala Alanine V =Val =Valine, L Leu Leucine I Ile Isoleucine 10 P =Pro =Proline *F Phe =Phenylalanine W Trp Tryptophan M Met Methionine *G GJly Glycine S Ser Serine T Thr Threonine C =Cys= Cysteine Y =Tyr =Tyrosine N Asn Asparagine Q Gin Glutamine D Asp =Aspartic Acid *E Glu. Glutamic Acid K K=Lys =Lysine Arg Arginine 2 H His Histidine

DEFINITIONS

The following terms are also used herein: The term "naturally occurring amino acids" means the L-isomers of the naturally occurring amino acids. The naturally occurring amino acids are glycine, alanine, valine, leucine, isoleucine, serine, methionine, threonine, phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine, aspartic 7 acid, asparagine, glutamic acid, glutamine, 7 -carboxyglutamic acid, arginine, ornithine and lysine.

Unless specifically indicated, all amino acids referred to in this application are in the L-form.

The term "unnatural amino acids" means amino acids that are not naturally found in proteins.

Examples of unnatural amino acids used herein, include racemic mixtures of selenocysteine and selenomethionine. In addition, unnatural amino acids include the D or L forms of nor-leucine, paranitrophenylalanine, homophenylalanine, para- S fluorophenylalanine, 3-amino-2-benzylpropionic acid, homoarginine, and D-phenylalanine.

SThe term "positively charged amino acid" 15 includes any naturally occurring or unnatural amino acid having a positively charged side chain under normal physiological conditions. Examples of positively charged naturally occurring amino acids are arginine, lysine and histidine.

20 The term "negatively charged amino acid" includes any naturally occurring or unnatural amino acid having a negatively charged side chain under normal physiological conditions. Examples of negatively charged naturally occurring amino acids are aspartic acid and glutamic acid.

The term "hydrophobic amino acid" means any amino acid having an uncharged, nonpolar side chain that is relatively insoluble in water. Examples of naturally occurring hydrophobic amino acids are alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and methionine.

The term "hydrophilic amino acid" means any amino acid having an uncharged, polar side chain that i is relatively soluble in water. Examples of naturally occurring hydrophilic amino acids are serine, infra, i.

I

-8threonine, tyrosine, asparagine, glutamine, and cysteine.

The term "mutant" refers to an ICE polypeptide, a polypeptide displaying the biological activity of wild-type, human ICE, characterized by the replacement of at least one amino acid from the wild-type, human ICE sequence according to Thornberry, N.A. et al., Nature, 356, pp. 768-774 (1992). Such a mutant may be prepared, for example, by expression of ICE cDNA previously altered in its coding sequence by oligonucleotide-directed mutagenesis.

SICE mutants may also be generated by site- S specific incorporation of unnatural amino acids into ICE proteins using the general biosynthetic method of 15 Noren, et al., Science, 244, pp. 182-188 (1989).

In this method, the codon encoding the amino acid of interest in wild-type ICE is replaced by a "blank" nonsense codon, TAG, using oligonucleotide-directed mutagenesis (described in detail, infra). A suppressor 20 tRNA directed against this codon is then chemically aminoacylated in vitro with the desired unnatural amino acid. The aminoacylated tRNA is then added to an in vitro translation system to yield a mutant ICE enzyme with the site-specific incorporated unnatural amino acid.

oa. Selenocysteine or selenomethionine may be incorporated into wild-type or mutant ICE by expression of ICE-encoding cDNAs in auxotrophic E. coli strains.

Hendrickson, W.A. et al., EMBO pp. 1665-1672 (1990). In this method, the wild-type or mutagenized ICE cDNA may be expressed in a host organism on a growth medium depleted of either natural cysteine or methionine (or both) but enriched in selenocysteine or selenomethionine (or both).

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9 The term "altered surface charge" means a change in one or more of the charge units of a mutant polypeptide, at physiological pH, as compared to wildtype ICE. This is preferably achieved by mutation of at least one amino acid of wild-type ICE to an amino acid comprising a side chain with a different charge at physiological pH than the original wild-type side chain.

The change in surface charge is determined by measuring the isoelectric point (pi) of the polypeptide molecule containing the substituted amino acid and comparing it to the isoelectric point of the wild-type ICE molecule.

The term "high specific activity" refers to a specific activity of ICE where the second-order rate constant (keat/Km) for hydrolysis of the substrate Ac- Tyr-Val-Ala-Asp-aminomethylcoumarin exceeds 7 x 104 M7' s 1 at 250C, using the assay described by Pennington, M.W. and N.A. Thornberry, Peptide Res., pp. 72- 20 76 (1994). Alternatively, the specific activity of ICE Bmay be determined by monitoring hydrolysis of the substrate Ac-Tyr-Val-Ala-Asp-p-nitroaniline. Reiter, Intr. J. Peptide Protein Res., 43, pp. 8796 I *(1994).

The term "altered substrate specificity" refers to a change in the ability of a mutant ICE to cleave a substrate as compared to wild-type ICE.

Substrate specificity may be measured by hydrolysis of fluorogenic peptide substrates or of unmodified peptide substrates by ICE, as described in Thornberry et al.suora. ICE mutants with altered substrate specificity demonstrate a second order rate constant (kcat/K.) for a substrate X 1 -Tyr-Val-Ala-X 2

-X

3 that exceeds the k ca/Km for the analogous peptide substrate X -Tyr-Val-Ala- Asp-X 3 For both substrates. X, is an amino protecting 9 The term "altered surface charge" means a change in one or more of the charge units of a mutant polypeptide, at physiological pH, as compared to wildtype ICE. This is preferably achieved by mutation of at least one amino acid of wild-type ICE to an amino acid comprising a side chain with a different charge at physiological pH than the original wild-type side chain.

The change in surface charge is determined by measuring the isoelectric point (pl) of the polypeptide II' molecule containing the substituted amino acid and comparing it to the isoelectric point of the wild-type ICE molecule.

The term "high specific activity" refers to a specific activity of ICE where the second-order rate constant (kcat/Km) for hydrolysis of the substrate Ac- Tyr-Val-Ala-Asp-aminomethylcoumarin exceeds 7 x 104 M' s1 at 25 0 C, using the assay described by Pennington, M.W. and N.A. Thornberry, Peptide Res., pp. 72- 20 76 (1994). Alternatively, the specific activity of ICE may be determined by monitoring hydrolysis of the substrate Ac-Tyr-Val-Ala-Asp-p-nitroaniline. Reiter, Intr. J. Peptide Protein Res., 43, pp. 8796 (1994).

S The term "altered substrate specificity" S refers to a change in the ability of a mutant ICE to cleave a substrate as compared to wild-type ICE.

Substrate specificity may be measured by hydrolysis of fluorogenic peptide substrates or of unmodified peptide substrates by ICE, as described in Thornberrv et al., supra. ICE mutants with altered substrate specificity Sdemonstrate a second order rate constant (kca/K for a substrate X -Tyr-Val-Ala-X 2

-X

3 that exceeds the kcat/Km for the analogous peptide substrate X 1 -Tyr-Val-Ala- 35 Asp-X 3 For both substrates, X 1 is an amino protecting AIo I group, such as acetyl; X 2 is a natural or unnatural amino acid residue other than L-aspartate; and X 3 is a carboxyl protecting group, such as aminomethylcoumarin or p-nitroaniline.

The "kinetic form" of ICE refers to the condition of the enzyme in its free or unbound form or bound to a chemical entity at either its active site or accessory binding site.

A "competitive" inhibitor is one that inhibits ICE activity by binding to the same kinetic form, of ICE, as its substrate binds thus directly competing with the substrate for the active site of S .ICE. Competitive inhibition can be reversed completely by increasing the substrate concentration.

15 An "uncompetitive" inhibitor is one that S inhibits ICE by binding to a different kinetic form of Sthe enzyme than does the substrate. Such inhibitors bind to ICE already bound with the substrate and not to the free enzyme. Uncompetitive inhibition cannot be 20 reversed completely by increasing the substrate concentration.

A "non-competitive" inhibitor is one that can bind to either the free or substrate bound form of ICE.

Those of skill in the art may identify inhibitors as competitive, uncompetitive or noncompetitive, by computer fitting enzyme kinetic data using standard equations according to Segel, I.H., Enzyme Kinetics, J...Wiley Sons, (1975). It should also be understood that uncompetitive o6inoncompetitive inhibitors according to this invention may bind to the accessory binding.ite.

The term "homologue" means a protein having at least 30% amino acid sequence identity with ICE or any functional domain of ICE as defined by Thornberry et al.. sunra and Casano et al., supra.

~--o~-rar~-Dsur 11 The term "subunit dissociation" refers to the fact that at very high dilutions of wild-type ICE, or at concentrations of the enzyme below 10 nM, enzymatic activity shows a time dependent loss assayed in the presence of a tetrapeptide substrate. Reconcentration of the dilute, inactive mixture results in complete recovery of ICE activity. Wild-type ICE demonstrates a Kd for subunit dissociation between 1 and 10 nM.

Enzymatic activity is determined by measuring the activity of ICE according to the assay of Penninqton Sand Thornberrv, supra, at varying concentrations of the enzyme. The concentration of the enzyme is determined by active site titration.

The term "co-complex" means ICE or a mutant 1. 5 or homologue of ICE in covalent or non-covalent association with a chemical entity or compound.

The term "associating with" refers to a condition of proximity between a chemical entity or compound, or portions thereof, and an ICE molecule or 20 portions thereof. The association may be noncovalent wherein the juxtaposition is energetically favored by hydrogen bonding or van der Waals or c\i electrostatic interactions or it may be covalent.

.,,The term "-sheet" refers to the conformation of a polypeptide chain stretched into an extended zigzig conformation. Portions of polypeptide chains that run "parallel" all run in the same direction.

SPolypeptide chains .that are "antiparallel" run in the opposite direction from the parallel chains.

The term "active site" or "active site moiety" refers to any or all of the following sites in ICE: the substrate binding site; the site where the tetrapeptide inhibitor binds and the site where the cleavage of a substrate occurs. The active site is characterized by at least amino acid residues 173, 176, 12 177, 178, 179, 180, 236, 237, 238, 239, 244, 248, 283, 284, 285, 290, 338, 339, 340, 341, 342, 343, 344, 345, 348, 352, 381 and 383, using the sequence and numbering according to Thornberrv et al., sura (SEQ ID NO:1).

The term "accessory binding site" or "accessory binding site moiety" refers to a binding site of ICE comprising amino acid residues adjacent to the two-fold axis of ICE but external to the active site, according to Table A. An accessory binding site may be a locus of ICE inhibition, although it is not the site of substrate cleavage.

The accessory binding site is characterized by at least amino acid residues 150, 151, 240, 259, 267, 268, 274, 291, 292, 293, 294, 295, 296, 297, 317, 318, 319, 320, 321, 322, 323, 324, 325, 327, 334, 335, 367, 371, 374, 375, 377, 378, 380, 382, 384, 386, 388, 389, 390, 391, 392, 393, 394, 395 and 396, using the sequence and numbering according to Thornberrv et al., supra (SEQ ID NO:1).

S 20 The term "P binding pocket" refers to a binding subsite, or portion of the binding site on the ICE molecule. The amino acid residues of an ICE substrate are given designations according to their position relative to the scissile bond, i.e. the bond that is broken by the protease. Residues are designated P1, P2, etc., for those extending toward the N-terminus from the scissile bond of the substrate.

The residues are designated P1', P2', etc., 'or those extending toward the C-terminus from the scissile bond of the substrate.

The portions of an ICE inhibitor that Scorrespond to the P or P' residues of the substrate are Salso labeled P1, P1', etc., by analogy with the a substrate. The binding subsites of the ICE molecule that receive the residues labeled P1, P1', etc., are 13 13 designated "the S1 site", "the P1' binding pocket", etc. Schechter, I. and A. Berger, "On the Size of the Active Site in Proteases", Biochem. Biophys. Res.

Commun., 27, pp. 157-162 (1967).

The "P1 binding pocket" of the ICE active site is defined as the space surrounded by amino acid residues Arg-179, His-237, Gln-283 and Arg-341.

The "P2 binding pocket" of the ICE active site is defined as the space surrounded by amino acid residues Pro-290, Val-338 and Trp-340.

'The "P3 binding pocket" of the ICE active site is defined as the space surrounded by amino acid S residues Pro-177, Arg-178, Thr-180, Arg-341 and f Pro-343.

The "P4 binding pocket" of the ICE active S site is defined as the space surrounded by amino acid residues Trp-340, His-342, Met-345, Val-348, Arg-352, Asp-381 and Arg-383.

The binding pocket" of the ICE active site is defined as the space surrounded by amino acid residues Phe-173, Ile-176, His-237, Gly-238, Ile-239, Cys-244 and His-248.

The term "p10 subunits interacting across the two-fold axis" means having at least 50% of the interface contacts according to Table A.

The term "structure coordinates" refers to mathematical coordinates derived from mathematical equations related to the patterns obtained on diffraction of a monochromatic beam of X-rays by the atoms (scattering centers) of an ICE molecule in crystal form. The diffraction data are used to calculate an electron density map of the repeating unit of the crystal. The electron density maps are used to 'establish the positions of the individual atoms within the unit cell of the crystal.

i *a 14 The term "heavy atom derivatization" refers to the method of producing a chemically modified form of a crystal of ICE. In practice, a crystal is soaked in a solution containing heavy metal atom salts, or organometallic compounds, lead chloride, gold thiomalate, thimerosal or uranyl acetate, which can diffuse through the crystal and bind to the surface of the protein. The location(s) of the bound heavy metal atom(s) can be determined by X-ray diffraction analysis of the soaked crystal. This information, in turn, is Sused to generate the phase information used to S construct three-dimensional structure of the enzyme.

SBlundel, T.L. and N.L. Johnson, Protein Crystallography, Academic Press (1976).

Those of skill in the art understand that a S set of structure coordinates determined by X-ray crystallography is not without standard error. For the purpose of this invention, any set of structure coordinates for ICE or ICE homologues or ICE mutants 20 that have a root mean square deviation of protein backbone atoms Ca, C and 0) of less than 0.75A when superimposed using backbone atoms on the St o structure coordinates listed in Table B shall be considered identical.

S 25 The term "unit cell" refers to a basic parallelipiped shaped block. The entire volume of a crystal may be constructed by regular assembly of such blocks. Each unit cell comprises a complete representation of the unit of pattern, the repetition of which builds up the crystal.

The term "space group" refers to the arrangement of symmetry elements of a crystal.

The term "molecular replacement" refers to a method that involves generating a preliminary model of an ICE crystal whose structure coordinates are unknown, 8 15 by orienting and positioning a molecule whose structure coordinates are known ICE coordinates from Table B) within the unit cell of the unknown crystal so as best to account for the observed diffraction pattern of the unknown crystal. Phases can then be calculated from this model and combined with the observed amplitudes to give an approximate Fourier synthesis of the structure whose coordinates are unknown. This, in turn, can be subject to any of the several forms of refinement to provide a final, accurate structure of the unknown crystal. Lattman, "Use of the Rotation and Translation Functions", in Methods in Enzymolov, 115, pp. 55-77 (1985); M.G. Rossmann, ed., "The Molecular Replacement Method", Int. Sci. Rev. Ser., No. 13, Gordon Breach, New York, (1972). Using the structure coordinates of ICE provided by this invention, molecular replacement may be used to determine the structure coordinates of a crystalline mutant or homologue of ICE or of a different crystal 20 form of ICE.

DETAILED DESCRIPTION OF THE INVENTION In order that the invention described herein may be more fully understood, the following detailed Sdescription is set forth.

The present invention relates to crystalline interleukin-l1 converting enzyme the structure of ICE as determined by X-ray crystallography, the use of that structure to solve the structure of ICE homologues and of other crystal forms of ICE, mutants and co-complexes of ICE, and the use of the ICE structure and that of its homologues, mutants and cocomplexes to design inhibitors of ICE.

*V.

I s r- i r~irPaarrrpr~ol 16- A. The Structure of ICE The present invention provides, for the first time, crystals of human ICE grown in the presence of a tetrapeptide inhibitor from solutions of polyethylene glycol, as well as the structure of ICE as determined therefrom. The crystals have tetragonal space group symmetry P4 3 2 1 2. The unit cell of said crystals has a rectangular shape of dimensions a=b=65 5A, and c=162 5A. The structure coordinates of ICE, as determined by X-ray crystallography of crystalline ICE, are listed in Table B.

Crystal packing reveals that ICE is a (p20) 2 /(pl0)2 tetramer. In the tetramer, two subunits contact two adjacent plO subunits which 15 interact across the crystallographic two-fold axis (Figure This axis corresponds to an oligomer interface in solution. Most of the dimer-dimer interface consists of p20 residues 291-297 and of residues 318-322 and 386-396.

20 Figure 1 represents a ribbon drawing of the p20/pl0 ICE heterodimer. As depicted in the figure, the p20 and p10 subunits are intimately associated and the active site is at the top of the figure, roughly at the center of the cluster of displayed side chains.

The enzyme core is a six-stranded B-sheet with 5 parallel strands (numbered 1, 2, 3, 4 and 7) and one anti-parallel strand (numbered Six a-helices (lettered A, B. E and F) lie roughly parallel to the S-strands. The last seven residues of p20 and the first seven of pl0 protrude from this compact structure and form two anti-parallel B-strands [5 (residues 291- 297)] and 6 (residues 317-323)1. A few key residues are labelled according to their position in the amino acid sequence of ICE (Thornberrv et al., sura).

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-17our understanding of the structure of ICE has enabled, for the first time, identification of the active site and accessory binding site of the enzyme.

The p10 subunit from one ICE molecule contacts the subunit from a different molecule and together they create an active site. The active site spans both the and p10 subunits and comprises amino acid residues from both subunits. The active site moiety is characterized by at least amino acid residues 173, 176, 177, 178, 179, 180, 236, 237, 238, 239, 244, 248, 283, 284, 285, 290, 338, 339, 340, 341, 342, 343, 344, 345, 348, 352, 381 and 383 using the sequence numbering according to Thornberrv et al-, surnra (SEQ ID NO: 1).

An accessory binding site is formed by amino 415 acid residues on the p10 subunits that interact across *the two-fold axis. The accessory binding site moiety is characterized by at least amino acid residues 150, 151, 240, 259, 267, 268, 274, 291, 292, 293, 294, 295, 296, 297, 317, 318, 319, 320, 321, 322, 323, 324, 325, 20 327, 334, 335, 367, 371, 374, 375, 377, 378, 378, 380, *382, 384, 386, 388, 389, 390, 391, 392, 393, 394, 395 and 396 using the sequence numbering according to Thornberry et al., supra (SEQ ID NO:l).

Uses of the Structure Coordinates of ICE ,25 For the first time, the present invention permits the use of molecular design techniques to design, select and synthesize chemical entities and compounds, including inhibitory compounds, capable of binding to the active site or accessory binding site of ICE, in whole or in part.

On approach enabled by this invention, is to use the structure coordinates of ICE to design compounds that bind to the enzyme and alter the physical properties of the compounds in different ways, is solubility. For example, this invention enables the design of compounds that act as competitive inhibitors of the ICE enzyme by binding to, all or a portion of, the active site of ICE. This invention also enables the design of compounds that act as uncompetitive inhibitors of the ICE enzyme- These inhibitors may bind to, all or a portion of, the accessory binding site of an ICE already bound to its substrate and may be more potent and less non-specific than known competitive inhibitors that compete only for the ICE active site. Similarly, non-competitive inhibitors that bind to and inhibit ICE whether or not it is bound to another chemical entity may be designed using the structure coordinates of ICE of this i invention.

A second design approach is to probe an ICE crystal with molecules composed of a variety of different chemical entities to determine optimal sites for interaction between candidate ICE inhibitors and 20 the enzyme. For example, high resolution X-ray diffraction data collected from crystals saturated with solvent allows the determination of where each type of solvent molecule sticks. small molecules that bind tightly to those sites can then be designed and synthesized and tested for their ICE inhibitor activity. Travis, _Science, 262, p. 1374 (1993).

This invention also enables the development of compounds that can isomerize to short-lived reaction intermediates in the chemical reaction of a substrate 30 or other compound that binds to ICE, with ICE. Thus, the time-dependent analysis of structural changes in ICE during its interaction with other molecules is enabled. The reaction intermediates of ICE can also be deduced from the reaction product in co-complex with ICE. Such information is useful to design improved 'M NMI -19analogues of known ICE inhibitors or to design novel classes of inhibitors based on the reaction intermediates of the ICE enzyme and ICE--inhibitor cocomplex. This provides a novel route for designing ICE inhibitors with both high specificity and stability.

Another approach made possible and enabled by this invention, is to screen compultationally small molecule data bases for chemical entities or compounds that can bind in whole, or in part, to the ICE enzyme.

In this screening, the quality of fit of such entities or compounds to the binding site may be judged either by shape complementarity or by estimated interaction energy. Meng, E.C. et J. Comp. Chem- 13, pp. 505-524 (1992).

Because ICE may crystallize in more than one crystal form, the structure coordinates of ICE, or portions thereof, as provided by this invention are particularly useful to solve the structure of those other crystal forms of ICE. They may also be used to 20 solve the structure of ICE mutants, ICE co-complexes, or of the crystalline form of any other protein with significant amtino acid sequence homology to any functional domain of ICE.

one method that may be employed for this purpose is molecular replacement. In this method, the 21 unknown crystal structure, whether it is another crystal form of ICE, an ICE mutant, or an ICE cocomplex, or the crystal of same other protein with significant amino acid sequence homology to any functional domain of ICE, may be determined using the ICE structure coordinates of this invention as provided in Table B. This method will provide an accurate structural form for the unknown crystal more quickly and efficiently than attempting to determine such information ab initio.

In addition, in accordance with this invention, ICE mutants may be crystallized in cocomplex with known ICE inhibitors. The crystal structures of a series of such complexes may then be solved by molecular replacement and compared with that of wild-type ICE. Potential sites for modification within the various binding sites of the enzyme may thus be identified. This information provides an additional tool for determining the most efficient binding interactions, for example, increased hydrophobic interactions, between ICE and a chemical entity or compound.

All of the complexes referred to above may be studied using well-known X-ray diffraction techniques 15 and may be refined versus 2-3A resolution X-ray data to an R value of about 0.20 or less using computer software, such as X-PL40R (Yale University, 01992, distributed by Molecular Simulations, Inc.). See, Blundel Johnson, supra; Methods in Enzv,-iolocrv.

20 vol. 114 115, H.W. Wyckoff et al., eds., Academic ~:.Press (1985). This information may thus be used to optimize known classes of ICE inhibitors, and more 9: importantly, to design and synthesize novel classes of ICE inhibitors.

The structure coordinates of ICE mutants provided in this invention also facilitate the identification of related proteins or enzymes analogous to ICE in function-, structure or both, thereby further leading to novel therapeutic modes for treating or preventing IL-I mediated diseases.

The design of compounds that bind to or inhibit ICE according to this invention generally involves consideration of two factors. First, the compound must be capable of physically and struicturally associating with ICE. Non-covalent molecular 21 interactions important in the association of ICE with its substrate include hydrogen bonding, van der Waals and hydrophobic interactions.

Second, the compound must be able to assume a conformation that allows it to associate with ICE.

Although certain portions of the compound will not directly participate in this association with ICE, those portions may still influence the overall conformation of the molecule. This, in turn, may have a significant impact on potency. Such conformational requirements include the overall three-dimensional structure and orientation of the chemical entity or compound in relation to all or a portion of the binding site, active site or accessory binding site of 15 ICE, or the spacing between functional groups of a compound comprising several chemical entities that directly interact with ICE.

The potential inhibitory or binding effect of a chemical compound on ICE may be analyzed prior to its 20 actual synthesis and testing by the use of computer modelling techniques. If the theoretical structure of the given compound suggests insufficient interaction and association between it and ICE, synthesis and testing of the compound is obviated. However, if computer modelling indicates a strong interaction, the molecule may then be synthesized and tested for its ability to bind to ICE and inhibit using the Sfluorescent substrate assay of Thornberrv et al., supra. In this manner, synthesis of inoperative compounds may be avoided.

An inhibitory or other binding compound of ICE may be computationally evaluated and designed by means of a series of steps in which chemical entities or fragments are screened and selected for their a- 22 ability to associate with the individual binding pockets or other areas of ICE.

One skilled in the art may use one of several methods to screen chemical entities or fragments for their ability to associate with ICE and more particularly with the individual binding pockets of the ICE active site or accessory binding site. This process may begin by visual inspection of, for example, the active site on the computer screen based on the ICE coordinates in Table B. Selected fragments or chemical entities may then be positioned in a variety of Sorientations, or docked, within an individual binding pocket of ICE as defined supra. Docking may be accomplished using software such as Quanta and Sybyl, 15 followed by energy minimization and molecular dynamics with standard molecular mechanics forcefields, such as CHARMM and AMBER.

Specialized computer programs may also assist in the process of selecting fragments or chemical 20 entities. These include: 1. GRID (Goodford, "A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules", J. Med. Chem., 28, pp. 849-857 (1985)). GRID is 25 available from Oxford University, Oxford, UK.

2. MCSS (Miranker, A. and M. Karplus, "Functionality i Maps of Binding Sites: A Multiple Copy Simultaneous Search Method." Proteins: Structure. Function and Genetics, 11, pp. 29-34 (1991)). MCSS is available from Molecular Simulations, Burlington, MA.

3. AUTODOCK (Goodsell, D.S_ and A.J. Olsen, "Automated Docking of Substrates to Proteins by Simulated Annealing", Proteins: Structure, Function, and Genetics, 8, pp. 195-202 (1990)).

AUTODOCK is available from Scripps Research Institute, La Jolla, CA.

4. DOCK (Kuntz, I.D. et al., "A Geometric Approach to Macromolecule-Ligand Interactions", J. Mol.

Biol., 161, pp. 269-288 (1982)). DOCK is -22 ability to associate with the individual binding pockets or other areas of ICE.

One skilled in the art may use one of several methods to screen chemical entities or fragments for their ability to associate with ICE and more particularly with the individual binding pockets of the ICE active site or accessory binding site. This process may begin by visual inspection of, for example, the active site on the computer screen based on the ICE coordinates in Table B. Selected fragments or chemical entities may then be positioned in a variety of orientations, or docked, within an individual binding pocket of ICE as defined supra. Docking may be accomplished using software such as Quanta and Sybyl, 15 followed by energy minimization and molecular dynamics with scandard molecular mechanics forcefields, such as CHARMM and AMBER.

Specialized computer programs may also assist in the process of selecting fragments or chemical entities. These include: 1. GRID (Goodford, "A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules", J. Med. Chem., 28, pp. 849-857 (1985)). GRID is available from Oxford University, Oxford, UK.

2. MCSS (Miranker, A. and M. Karplus, "Functionality Maps of Binding Sites: A Multiple Copy Simultaneous Search Method." PRoteins: Structure, Function and Genetics, 11, pp. 29-34 (1991)). MCSS is available from Molecular Simulations, Burlington, MA.

3. AUTODOCK (Goodsell, D.S. and A.J. Olsen, "Automated Docking of Substrates to Proteins by Simulated Annealing", Proteins: Structure, Function, and Genetics, B, pp. 195-202 (1990)).

AUTODOCK is available from Scripps Research Institute, La Jolla, CA.

4. DOCK (Kurtz, I.D. et al., "A Geometric Approach to Macromolecule-Ligand Interactions", J. Mol.

Biol., 161, pp. 269-288 (1982)). DOCK is 23 available from University of California, San Francisco, CA.

Once suitable chemical entities or fragments have been selected, they can be assembled into a single compound or inhibitor. Assembly may be proceed by visual inspection of the relationship of the fragments to each other on the three-dimensional image displayed on a computer screen in relation to the structure coordinates of ICE. This would be followed by manual model building using software such as Quanta or Sybyl.

Useful programs to aid one of skill in the art in connecting the individual chemical entities or fragments include: 1. CAVEAT (Bartlett, P.A. et al, "CAVEAT: A Program 15 to Facilitate the Structure-Derived Design of Biologically Active Molecules". In Molecular Recocnition in Chemical and Biological Problems", Special Pub., Royal Chem. Soc., 78, pp. 182-196 (1989)). CAVEAT is available from the University of California, Berkeley, CA.

2. 3D Database systems such as MACCS-3D (MDL Information Systems, San Leandro, CA). This area is reviewed in Martin, "3D Database Searching in Drug Design", J. Med. Chem., pp. 2145-2154 (1992)).

3. HOOK (available from Molecular Simulations, Burlington, MA).

Instead of proceeding to build an ICE inhibitor in a step-wise fashion one fragment or chemical entity at a time as described above, inhibitory or other ICE binding compounds may be designed as a whole or "de novo" using either an empty active site or optionally including some portion(s) of a known inhibitor(s). These methods include: 1. LUDI (Bohm, "The Computer Program LUDI: A New Method for the De Novo Design of Enzyme Inhibitors", J. Como. Aid. Molec. Design, 6, pp. 61-78 (1992)). LUDI is available from Biosym Technologies, San Diego, CA.

24 2. LEGEND (Nishibata, Y. and A. Itai, Tetrahedron, 47, p. 8985 (1991)). LEGEND is available from Molecular Simulations, Burlington, MA.

3. LeapFrog (available from Tripos Associates, St. Louis, MO).

Other molecular modelling techniques may also be employed in accordance with this invention. See, Cohen, N.C. et al., "Molecular Modeling Software and Methods for Medicinal Chemistry, J. Med. Chem., 33, pp. 883-894 (1990). See also, Navia, M.A. and M.A.

Murcko, "The Use of Structural Information in Drug Design", Current Opinions in Structural Biolocyv 2, pp. 202-210 (1992).

Once a compound has been designed or selected 15 by the above methods, the efficiency with which that compound may bind to ICE may be tested and optimized by I' computational evaluation. For example, a compound that has been designed or selected to function as an ICEinhibitor must also preferably traverse a volume not 20 overlapping that occupied by the active site when it is "bound to the native substrate. An effective ICE inhibitor must preferably demonstrate a relatively small difference in energy between its bound and free S" states a small deformation energy of binding).

25 Thus, the most efficient ICE inhibitors should preferably be designed with a deformation energy of binding of not greater than about 10 kcal/mole, preferably, not greater than 7 kcal/mole. ICE inhibitors may interact with the enzyme in more than one conformation.that is similar in overall binding energy. In those cases, the deformation energy of binding is taken to be the difference between the energy of the free compound and the average energy of the conformations observed when the inhibitor binds to the enzyme.

Ji m 25 A compound designed or selected as binding to ICE may be further computationally optimized so that in its bound state it would preferably lack repulsive electrostatic interaction with the target enzyme. Such non-complementary electrostatic) interactions include repulsive charge-charge, dipole-dipole and charge-dipole interactions. Specifically, the sum of all electrostatic interactions between the inhibitor and the enzyme when the inhibitor is bound to ICE, preferably make a neutral or favorable contribution to the enthalpy of binding.

Specific computer software is available in the art to evaluate compound deformation energy and electrostatic interaction. Examples of programs 15 designed for such uses include: Gaussian 92, revision C Frisch, Gaussian, Inc., Pittsburgh, PA 01992]; AMBER, version 4.0 Kollman, University of California at San Francisco, 01994]; QUANTA/CHARMM [Molecular Simulations, Inc., Burlington, MA 01994]; S 20 and Insight II/Discover (Biosysm Technologies Inc., San Diego, CA 01994). These programs may be implemented, for instance, using a Silicon Graphics workstation, IRIS 4D/35 or IBM RISC/6000 workstation Stod, model 550. Other hardware systems and software packages will be known to those skilled in the art.

Once an ICE-binding compound has been optimally selected or designed, as described above, substitutions may then be made in some of its atoms or side groups in order to improve or modify its binding properties. Generally, initial substitutions are conservative, the replacement group will have approximately the same size, shape, hydrophobicity and charge as the original group. It should, of course, be understood that components known in the art to alter conformation should be avoided. Such substituted IN

IN

26 chemical compounds may then be analyzed for efficiency of fit to ICE by the same computer methods described in detail, above.

C. Mutants Of ICE The present invention also enables mutants of ICE and the solving of their crystal structure. More particularly, by virtue of the present invention, the location of the active site, accessory binding site and interface of ICE based on its crystal structure permits S* 10 the identification of desirable sites for mutation.

For example, mutation may be directed to a particular site or combination of sites of wild-type SICE, the accessory binding site or only the active site, or a location on the interface site may be S 15 chosen for mutagenesis. Similarly, only a location on, at or near the enzyme surface may be replaced, resulting in an altered surface charge of one or more charge units, as compared to the wild-type enzyme.

'0 1 Alternatively, an amino acid residue in ICE may be 20 chosen for replacement based on its hydrophilic or hydrophobic characteristics.

Such mutants may be characterized by any one of several different properties as compared with wildtype ICE. For example, such mutants may have altered 25 surface charge of one or more charge units, or have an increased stability to subunit dissociation. Or such mutants may have an altered substrate specificity in comparison with, or a higher specific activity than, wild-type ICE.

The mutants of ICE prepared by this invention may be prepared in a number of ways. For example, the wild-type sequence of ICE may be mutated in those sites identified using this invention as desirable for mutation, by means of oligonucleotide-directed -i -27 mutagenesis or other conventional methods, e.g.

deletion. Alternatively, mutants of ICE may be generated by the site specific replacement of a particular amino acid with an unnaturally occurring amino acid. In addition, ICE mutants may be generated through replacement of an amino acid residue, or a particular cysteine or methionine residue, with selenocysteine or selenomethionine. This may be achieved by growing a host organism capable of expressing either the wild-type or mutant polypeptide on a growth medium depleted of either natural cysteine or methionine (or both) but enriched in selenocysteine or selenomethionine (or both).

Mutations may be introduced into a DNA S 15 sequence coding for ICE using synthetic oligonucleotides. These oligonucleotides contain nucleotide sequences flanking the desired mutation sites. Mutations may be generated in the full-length DNA sequence of ICE (p45) or in any sequence coding for p30, or p20 or pl0 polypeptides.

According to this invention, a mutated ICE DNA sequence produced by the methods described above, or any alternative methods known in the art, can be .expressed using an expression vector. An expression 25 vector, as is well known in the art, typically includes elements that permit autonomous replication in a host cell independent of the host genome, and one or more phenotypic markers for selection purposes. Either prior to or after insertion of the DNA sequences surrounding the desired ICE mutant coding sequence, an expression vector also will include control sequences encoding a promoter, operator, ribosome binding site, translation initiation signal, and, optionally, a repressor gene or various activator genes and a signal for termination. In some embodiments, where secretion l J :r j- -CL~i~ diiai 28 of the produced mutant is desired, nucleotides encoding a "signal sequence" may be inserted prior to the ICE mutant coding sequence. For expression under the direction of the control sequences, a desired DNA sequence must be operatively linked to the control sequences they must have an appropriate start signal in front of the DNA sequence encoding the ICE mutant and maintaining the correct reading frame to permit expression of that sequence under the control of the control sequences and production of the desired product -encoded by that ICE sequence.

Any of a wide variety of well known available S expression vectors are useful to express the mutated ICE coding sequences of this invention.

These include, for example, vectors consisting of segments of chromosomal, non-chromosomal and synthetic DNA sequences, such as various known derivatives of SV40, known bacterial plasmids, e.g., plasmids from E. coli including col El, pCRl, pBR322, pMB9 and their derivatives, wider host range plasmids, RP4, phage DNAs, the numerous derivatives of phage X, NM 989, and other DNA phages, e.g., M13 and filamentous single stranded DNA phages, yeast plasmids such as the 2( plasmid or derivatives thereof, :25 and vectors derived from combinations of plasmids and phage DNAs, such as plasmids which have been modified to employ phage DNA or other expression control sequences. In the.preferred embodiments of this invention, we employ E. coli vectors.

In addition, any of a wide variety of expression control sequences sequences that control the expression of a DNA sequence when operatively linked to it may be used in these vectors to express the mutated DNA sequences according to this invention.

Such useful expression control sequences, include, for 29 example, the early and late promoters of SV40 for animal cells, the lac system, the trp system the TAC or TRC system, the major operator and promoter regions of phage X the control regions of fd coat protein, all for E. coli, the promoter for 3-phosphoglycerate kinase or other glycolytic enzymes, the promoters of acid phosphatase, Pho5, the promoters of the yeast a-mating factors for yeast, and other sequences known to control the expression of genes of prokaryotic or eukaryotic cells or their viruses, and various combinations thereof. In the preferred embodiments of this invention, we employ either E. coli or eukaryotic .expression in COS-1 cells, a monkey kidney cell line.

A wide variety of hosts are also useful for 4 15 producing mutated ICE according to this invention.

These hosts include, for example, bacteria, such as E. coli, Bacillus and Streptomyces, fungi, such as yeasts, and animal cells, such as CHO and COS-1 cells, plant cells and transgenic host cells. In preferred embodiments of this invention, the host cells are E cli or COS-1 cells.

It should be understood that not all expression vectors and expression systems function in the same way to express mutated DNA sequences of this S 25 invention and to produce modified ICE or ICE mutants.

Neither do all hosts function equally well with the same expression system. However, one of skill in the art may make a selection among these vectors, expression control sequences and hosts without undue experimentation and without departing from the scope of this invention. For example, an important consideration in selecting a vector, will be the ability of the vector to replicate in a given host.

The copy number of the vector, the ability to control that copy number, and the expression of any other 'I

MUMEM

30 proteins encoded by the vector, such as antibiotic markers, should also be considered.

In selecting an expression control sequence, a variety of factors should also be considered. These include, for example, the relative strength of the system, its controllability, its compatibility with the DNA sequence encoding the modified ICE of this invention, particularly with regard to potential secondary structures.

Hosts should be selected by consideration of their compatibility with the chosen vector, the toxicity of the modified ICE to them, their ability to secrete mature products, their ability to fold proteins correctly, and to form tetramers, their fermentation requirements, the ease of the purification of the modified ICE from them and safety. Within these parameters, one of skill in the art may select various vector/expression control system/host combinations that will produce useful amounts of the mutant ICE.

The mutant ICE produced in these systems may be purified by a variety of conventional steps and strategies, including those used to purify wild-type

ICE.

Once the ICE mutants have been generated in the desired location, active site or accessory binding site, the mutants may be tested for any one of several.properties of interest.

For example, mutants may be screened for an altered charge at physiological pH. This is determined by measuring the mutant ICE isoelectric point (pI) in comparison with that of the wild-type parent.

Isoelectric point may be measured by gelelectrophoresis according to the method of Wellner, D., Analvt. Chem, 43, p. 597 (1971). A mutant with an altered surface charge is an ICE polypeptide containing r ~k~nrrsr~*36~clP~l~rapyppaypu 31 a replacement amino acid located at the surface of the enzyme, as provided by the structural information of this invention, and an altered pl.

Furthermore, mutants may be screened for high specific activity in relation to the wild-type ICE. A mutant would demonstrate high specific activity if its second order rate constant (K ,t/Km) for hydrolysis of the substrate Ac-Tyr-Val-Ala-Asp-amino methylcoumarin exceeds 7 x 104 M- 1 s at 25 0 C, using the assay in Pennington Thornberry, supra.

A mutant would be tested for altered ICE substrate specificity by measuring the hydrolysis of S.fluorgenic peptide substrates or unmodified ICE peptide substrates as described in Thornberry et al., supra.

An enzyme with altered substrate specificity is an enzyme whose second order rate constant (kca/Km) for a substrate X 1 -Tyr-Val-Ala-X 2 -X that exceeds the k at/Km for the analogous peptide substrate X 1 -Tyr-Val-Ala- Asp-X 3 X, is an amino protecting group, such as acetyl; X 2 is a natural or unnatural amino acid residue other than L-aspartate; X 3 is a carboxyl protecting group such as aminomethylcoumarin or p-nitroaniline.

Further properties of interest also include mutants with increased stability to subunit dissociation. An ICE mutant with increased stability to subunit dissociation would demonstrate no loss of enzymic activity at concentrations of the enzyme below nM in comparison with the wild-type ICE, which demonstrates a Kd between 1-10 nM.

In order that the invention described herein may be more fully understood, the following examples are set forth. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting this invention in any manner.

I 1MO11 NN 32 EXAMPLE 1 Crystal Structure of ICE The cDNA encoding the p30 precursor of active human ICE (residues Asn 120 to His 404 of (Thornberry et al., supra]) was cloned into a PL promoter expression vector (provided by Dr. J.

Mankovich) and expressed in E. coli by temperatureshift induction.

Pre-induction repression of the PL promoter 1. 0 was achieved by co-expression of the cl repressor gene on a co-resident, compatible plasmid (pACYC184cI) in the E. coli host, JM109. Yanish-Perron, et al., Gene, 33, pp. 103-199 (1985; ATCC #53323). The S promoter was induced by increasing the temperature from S 15 28 0 C to 420C, at which point the temperature sensitive c repressor gene product denatures and gene expression is initiated, directed by the PL promoter. Maintenance of the temperature at 42 0 C for a further 4 hours resulted in the accumulation of high levels of the 20 inactive ICE p30 precursor product within the host cell S cytoplasm, in the Hfrm of inactive inclusion bodies.

After mechanical disruption of the cells, and harvesting of the insoluble fraction, the inclusion bodies were washed by suspension in 2M urea, 25mM tris, 0.5mM DTT, 0.1mM EDTA, 0.1mM PMSF, pH 7.5 at 4oC, followed by centrifugation. The inclusion bodies were solubilized in the-above buffer containing 7M urea, centrifuged and subjected to size-exclusion chromatography in the same buffer. The p30 fractions, identified by SDS-PAGE and N-terminal sequence analysis, were pooled and diluted to 0.3 mg/ml using column buffer. This was followed by dialysis at against 25mM tris, 1mM DTT, pH 7.5, until the urea ~aPP~8s~e~B~Pcs~ 33 concentration was less than 20mM, thereby allowing the enzyme to refold.

The protein was concentrated to 3-5 mg/ml by ultrafiltration at 4 0 C, followed by incubation at room temperature. The disappearance of the p30 precursor, and the concomitant appearance of the p20 and plo subunits, was monitored by SDS-PAGE, evidence that autocatalytic processing of the enzyme had occurred.

ICE enzymatic activity was assayed by hydrolysis of a Succinyl-Tyr-Val-Ala-Asp-p-nitroanilide substrate at 37 0 C and correlated closely with conversion to subunits.

The autoprocessed ICE was inhibited fully by adding a 2x molar excess of a tetrapeptide aldehyde 15 inhibitor (acetyl-Tyr-Val-Ala-Asp-H). The protein-inhibitor complex sample was concentrated and fractionated by size-exclusion chromatography, in final preparation for crystallization experiments.

Crystals of ICE in complex with the inhibitor were grown by vapor diffusion. Davies, D.R. and D.M.

Segal, Meth. Enzvmol., 22, p. 266 (1971). Protein mg/ml in 50 mM citrate, 2.0 mM DTT, pH 6.5) was mixed Sl with an equal volume of reservoir buffer (15% PEG 4K, 400 mM LiSO4. 200 mM sodium Hepes, 5mM sodium cacodylate, 0.5% beta-octyl glucoside, pH 7.0) and allowed to stand over the reservoir solution at 4 0

C.

Crystals grew over a six week period to form tetragonal bipyramids and were equilibrated with 18% PEG 4K, 400 mM LiSO 4 200 mM sodium Hepes, 5mM sodium cacodylate, 0.5% beta-octyl glucoside, pH 7.0 prior to data collection or heavy atom derivatization.

Those of skill in the art will appreciate that the aforesaid crystallization conditions can be varied. Such variations may be used alone or in combination, and include final protein/inhibitor 34 complex concentrations between 5 mg/ml and 35 mg/ml; all combinations of ICE/inhibitor to precipitant ratios; citrate concentrations between 1mM and 200 mM; DTT concentrations between 0 mM and 10 mM; and any concentration of 9-mercaptoethanol; pH ranges between and 9.5; PEG concentrations between 10% and (gms/100ml); PEG weights between 2000 and 8000; LiSO 4 concentrations between 50 and 750 mM; HEPES concentrations between 5 and 395 mM; and any concentration or type of detergent; any temperature between -5°C and 30=C; and crystallization of ICE/inhibitor complexes by batch, liquid bridge, or dialysis method using these conditions or variations thereof.

All X-ray data sets were collected on a S, R-axis IIC image plate system except for the 2.2A Synchrotron data set that was used for refinement of the final model. This data was collected at Cornell High Energy Synchrotron Source ("CHESS") on a chargecouple device and was reduced to structure factor amplitudes using the Denzo Software Package (Denzo An Oscillation Data Processing Program For Macro Molecular Crystallography, 01993, Daniel Gewirth, Yale University). Oscillation photographs were integrated and reduced to structure factor amplitudes using software supplied by the manufacturer (Molecular Y C Structures Corp., Dallas, Texas).

Refined heavy atom parameters were used to compute multiple isomorphous replacement phases.

Inclusion of the anomalous data for the Hg derivative in cross-phased difference Fourier maps showed the space group to be P4 3 2 1 2 rather than its enantiomorph.

The mean figure of merit, including anomalous data for the Hg derivative, was 0.65 to 3.5K resolution (Table 1).

Solvent flattening and phase extension (CCP4- Collaborative Computing Project No. 4, A Suite of Programs for Protein Crystallography; Daresbury Laboratory, Warrington, WA4 4AD, U.K. (1979)) improved the map and allowed identification of some of the residues in the protein core-. Cycles of model buildingi (Quanta, version 4.0Ob, Molecular Simulations Inc., Burlington MA), positional refinement, (Brunger, A.T., J. Acta Cryst., A46, pp. 46-S7 (1990); Brunger, A.T.

et J. Acta Cryst., A46, pp. 585-93 (1990)) and phase combination (CCP4-Collaborative Computing Project, sunra) were carried out until the switch to phases calculated from the model could be made.

Refinement continued against the -16 0 C, 2.2A data (Table which allowed the more difficult loop regions of the protein to be constructed.

*The following table summarizes the X-ray crystallography data sets of ICE derivatives that were used to determine the structure of ICE according to this invention.

25 30 Table 1 C-ViewUfft Ca Resardw o Rmag. &camuioww A No. ot Rc phaaing PC4nwdcndeft. %K %b C sit" %K Poweri Tetuapetde aldetlyde* Z0- 2.2 87 7.1 64.9 164.1 1.tapvpt6 a&dsh'de* .20 -2.8 so 8.3 64.4 1633 Yetrtoeptdo a~dejd 20 -2.8 78 8.3 64.7 182.9 hIotnated tavapaptift a"d1y" 20 -3.5 86 3.4 64.4 162.5 2 0.88 1.09 20-3K 88 8RA 64.4 162.3 5, 0.61 1.03 Gold Thiate 20) 3.5 74 9I5 64.7 162-7 3 0.72 1.22 ~tkrtAcaett. 04 ea 10.a 64.7 162.9 2 0.75 1.3 Lead Otknuide 20.- 3A 6 8.9 6A-7 162-8 2 0.76 1.39 'atco&I lected at -16' C a, CHIESS Oata Colectd at -116C Definitions: Rmerge gives the agreement between repeated intensity measurements, with the number of crystals used in the data set given in parentheses.

The number of heavy-atomn binding sites is given where Solvent flattening and phase extension (CCP4- Collaborative Computing Project No. 4, A Suite of Programs for Protein Crystallography; Daresbury Laboratory, Warrington. WA4 4AD, U.K. (1979)) improved the map and allowed identification of some of the residues in the protein core. Cycles of model building (Quanta, version 4.0b, Molecular Simulations Inc., Burlington MA), positional refinement, (Brunger, A.T., J. Acta Crvst., A4G, pp. 46-57 (1990); Brunger, A.T.

et al., J. Acta Crvst., A46, pp. 585-93 (1990)) and phase combination (CCP4-Collaborative Computing Project, sup~ra) were carried out until the switch to phases calculated from the model could be made.

Refinement continued against the -16 0 C, 2.2A. data (Table which allowed the more difficult loop regions of the protein to be constructed.

*The following table summarizes the X-ray crystallography data sets of ICE derivatives that were used to determine the structure of ICE according to this invention.

25 *0 30 TabLe I ProRei Moiiain Aa~ton r~ssof fmRn*. din nhlon. A No. of Ac Phasing PrtenMofiale I ab% ,b a altes Power- Tetrapeptide aldehyde* 20 -2.2 87 7.1 64.9 184.1 Tetrapeptide aldshydae 20 -2.6 90 8.3 64.4 163.3 Tetrapaptlde aldishyde 20. 2.5 78 8.3 64.7 '162.9 lodinated tatrapaptide uldehyde 20- 3.5 86 9.4 64.4 162.8 2 0.88 1.09 Thimerosal 20. 3.5 as 8.4 64.4 162.3 5 0.67 1.08 Gold Thiomalate 20 -3.5 74 9.5 64.7 162.7 3 0.72 1.22 Ufan~q Acetate 20 4:0 so 10.8 64.7 162.9 2 0.79 1.32 Lead Chloride .20- 3.5 64 8.9 64.7 162.8 2 0.76 1.38 *Date collected at -16*C at CHESS Data collected at -14 Definitions: Rmerge gives the agreement between repeated intensity measurements, with the number ofA crystals used in the data set given in parentheses. I The number of heavy-atom binding sites is given where -36appropriate. R is the Cullis R factor for centrosynifetric reflections, and the phasing power is the ratio of average heavy-atom scattering to the average lack of closure of the phase triangles.

T.L. and Johnson, Protein Crystalloaa~hv, Academic Press, New York (1976).

The ICE tetrameric model according to this invention has an R-f actor of 19V against all observed data between 7A and 2.2A resolution, with root-mean-square deviation from ideal bond lengths and angles of 0.011A and 2.84A respectively.

_EXAM~PLE 2 Confirmation of the Active Site of ICE In order to confirm the location of the active site in the tetrameric ICE molecule, as deduced from the structure coordinates of ICE, a series of ICE mutants was generated.

Oligonucleotide-directed mutagenesis was performed on pcDNA3 (Invitrogen) constructs using v' 20 uracil-enrichment of single-strand DNA. Kunkel, T.A., Proc. Nat. Acad. Sci. 488-492 (1985); Kunkel, T.A. et al.- Meth. EnF mol., 154, pp. 367-382 (1987).' This I s a modif ication of the method originally described for M13 mutagenesis. Zoller, M.J. and M. Smith, Nucleic Acids Res-, 10, pp. 6487-6500 (1983); Zoller M.J. and M. Smith, Meth. Enzv-mol., 100, pp. 468- 500 (1983).

MutagenesiS was performed using the reagents *provided in the Muta-Gene Kit (BioRad). Mutagenesis primers were synthesized in the W+ coding orientation.

The dUtulcfE coli strain CJ326 was used for uracil enrichment of single-strand DNA, and the MV11.90 strain was used for selection of heteroduplex DNA after extension-ligatio. reactions. Aloligonuceoties f 37 were synthesized on an applied Biosystems 380 DNA synthesizer and purified by electrophoreses in polyacrylamide-urea slab gels. Mutations made in the ICE-encoding cDNA were fully sequenced in the coding region by the dideoxy method. Sanger, F.

et al., Proc. Nat. Acad. Sci. 74, pp. 5463-5467 (1977).

Mutant DNA in preparation for COS-1 cell transfection, or alternatively E. coli transfection, was purified by alkaline lysis and cesium gradient centrifugation prior to transfection.

Each mutant cDNA was transfected into a COS- 1 cell line, then tested for its ability to process pIL-19 in vitro, to secrete mature IL-1S. The COS-1 cell line used, had previously been transfected with aLpIL-l1 encoding cDNA cloned into an MNC stuffer vector Seed, Harvard Medical School) which had subsequently integrated into the chromosome. pIL-19 production was maintained by the addition of 0.5 mg/ml G-418 Sulfate to culture media.

Approximately 3 x 10 6 COS-1 cells in 100mm tissue culture plates were transfected with 15 gg of each plasmid. DNA was mixed with 200 Al DEAE-Dextran, brought to 4 ml with phosphate-buffered saline, and added to the plates. Cells were incubated at 37 0 C for S 25 30 min. 8 ml of an 80 AM chloroquine/serum-free DMEM solution was added and the cells were incubated for 2.5 hr.. This solution was aspirated and cells were treated for two minutes with 10% DMSO/serum-free DMEM.

After washing with serum-free media, 10 ml complete media was added. Conditioned media were sampled at 16 and 24 hr. Activity in this assay requires that transcription, translation and protein folding of mutants are not arrested. The amount of mutant ICE present in cell lysates was determined by Western blot using an anti-p20 rabbit antiserum that recognizes -38 amino acids 136-150 inclusive and which also recognizes the intact p30 precursor.

Mature IL-l1 in the cell medium was detected by ELISA (R&D Systems). Samples were diluted to achieve concentrations in the linear range of the ELISA assay (8-60 pg/ml). Background IL-1S levels were determined in cells transfected with the expression vector lacking ICE cDNA, and this value was subtracted from all other concentrations. The activity values were calculated as the ratio of secreted IL-1l from cells transfected with mutant ICE divided by IL-li secreted by cells transfected with wild-type ICE. The final ratio is the mean of at least two experiments.

S These data are recorded in Figure 3.

Based on these data, it was determined that mutation of Cys-285 or His-237 eliminates pIL-1 processing activity, as well as autoprocessing.

Mutation of Arg-179, which contacts the P1 Asp to Glu, also abolishes activity. Mutation of Cys-244 to Ala, which may contact P' side chains of substrates, reduces enzymatic activity significantly. In contrast, mutation of other residues proximal to Cys-285 including Ser-332, -333 or -339, and His-249, does not eliminate activity. Accordingly, we confirmed the importance of various residues in the ICE active site.

B S EXAMPLE 3 SThe Use of Molecular Replacement To Solve An Unknown ICE Crystal Structure The method of molecular replacement was used to determine the structure coordinates of crystals of ICE in complex with the tetrapeptide aldehyde inhibitor Ac-Tyr-Val-Pro-Asp-H in comparison with crystals of ICE in complex withthe tetrapeptide aldehyde inhibitor Ac-Tyr-Val-Ala-Asp-H (as prepared in Example 1).

i 39 -Crystals of ICE in complex with the tetrapeptide aldehyde inhibitor, Ac-Tyr-Val-Pro-Asp-H were grown under conditions identical to those for crystals of ICE in complex with the tetrapeptide aldehyde inhibitor, Ac-Tyr-Val-Ala-Asp-H X-ray diffraction data to 2.8A resolution was collected on the ICE/Pro co-complex. A difference electron density map that combined diffraction data of the form Fg FAla I and phases calculated from the refined model of the Ala inhibited enzyme was used to locate structure changes that had occurred in the ICE/Pro co-complex.

Negative features were found in the map wherever localized atoms in the Ala complex were 15 removed or shifted by switching to the new ligand.

Positive features were found when localized atoms were see introduced into the structure, and indicated the new positions of shifted atoms.

Replacement of the alanine that sits in the P2 binding pocket in Ala with proline in Pro introduced two methylene groups into the structure of the ICE cocomplex. The location of these new atoms was indicated Sby the presence of positive difference electron density adjacent to the beta-carbon of the alanine in the 25 binding pocket P2. Another positive peak nearby "t indicated the binding of a new water molecule in the Pro complex relative to the Ala complex. There were also pairs of positive and negative peaks near the tyrosine moiety that sits in the P4 binding pocket of the inhibitor. These peaks indicated shifts in the position of these atoms in the Pro complex relative to their location in the Ala complex.

These shifts, plus the new atoms referred to above, were modeled, and the resulting structure was refined against the X-ray data to determine a final i ;i I 40 picture of the co-complex of Pro with ICE. The space group (P4 3 2 1 2) and unit cell dimensions (a=b=65 c=162 5A) for the Pro complex were the same as those observed for the Ala complex.

The ICE structure coordinates known for the first time by virtue of this invention may be used to solve the unknown structure of any mutant, homologue or co-complex of ICE using the above-described method.

This method may also be used to determine the binding or orientation of a ligand or chemical entity in the active site or accessory binding site of ICE.

While we have described a number of embodiments of this invention, it is apparent that our 9 basic examples may be altered to provide other 15 embodiments which utilize the products and processes of this invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments which have been represented by way of example.

Tables A and B, following this page, list respectively, the tetramer interface contacts and the structure coordinates of the ICE molecule of this u invention.

S

V-

-t -U-mmlmm -41- TAMLE A TETRMER ITE~RFACE CONTACTS Residue Residue Residue P10 plo PlO) P20 150 375 320 382 240 259 151 371 320 380 267 293 151 372 322 377 268 293 151 375 322 378 274 295 291 323 322 380 291 321 322 384 291 322 322 385 291 323 322 386

I

292 321 323 327 293 321 324 334 15 293 319 324 386 293 320 325 378 -294 318 325 386 '294 319 334 393 *294 320 335 391 295 318 367 367 295 319 367 374 .295 320 371 394 295 321 371 395 ~:296 317 371 396 297 317 374 392 p374 393 pp374 394 .9.qpp375 395 375 396 378 395 378, 396 386 393 386 395 388 392 388 391 388 393 389 392 389 391 390 391 i L i r ii i

~I

a~ ;3:a.

42 TABLE B STRUCTURE COORDINATES OF ICE Atom Ii~ RESODUE I ATOM I C GLY 131 ATOM 2 0 GLY 131 ATOM 3 IITI GLY 131 ATOM 4 HT2 GLY 131 ATOM S N GLY 131 ATOM 6 HT3 .LY 131 ATOM 7 CA (3LY 131 ATOM B N ASN 132 ATOM 9 H ASN 132 ATOM 10 CA ASN 132 ATOM 13 CR ASN 132 V. ATOM 12 CG ASN 132 ATOM 13 OD1 ASN 132 ATOM 14 ND2 ASN 132 ATOM IS HD21 ASN 132 20 ATom 16 HD22 ASN 132 *&use: ATOM 17 C ASN 132 ATOM 18 0 ASN 132 ATOM 19 N VAL 133 ATOM 20 H VAL 133 25 ATOM 21 CA VAL 133 ATOM 22 CB VA. 133 ATOM 23 CGI VA. 133 ATOM 24 CG2 VAL 133 ATOM 25 C VAL 133 ATOM 26 0 VAL 133 ATOM 27 N LYS 134 ATOM 21 NZ LYS 134 ATOM 29 CA LYS 134 ATOM 30 CB LYS 134 ATOM 31 CG LYS 134 ATOM 32 CD LYS 134 ATOM 33 CE LYS 134 a. ATOM 34 HZ -LUS 134 ATOM 35 HZIi LYS 134 ATOM 36 HZ2 LYS 134 ATOM 37 1123 LYS 134 ATOM 38 C LYS 134 -a ATOM 39 0 LYS 134 ATOM 40 N LET) 135 ATOM 41 H LEU 135 ATOM 42 CA LEU 135 ATOM 43 CS LEU 13S ATOM 44 CG LEU 13S ATOM 45 CDI LEU 135 ATM 46 CD2 LEO 135 ATOl 47 C 1e" 135 AT~l 44 0 LEO 135 ATOM 49 H CYS 136 ATOM $0 1 CYS 136 ATOM 51 CA CYS 336 ATOM SZ C CYS 136 AtO 33 a CyI 13 ATVOM 34 CB CTS 136 ATOM 55 SG CYS 136 X y 7 (CC B 49."a1 81.52 30.205 82.686 51.316 82.385 49.746 82.510 50.546 81.841 50.783 81.456 50.192 80.805 49.175 80.916 4-1.640 80.121 49.178 81.466 47.778 81.260 47.550 82.132 48.228 83.107 46.506 81.860 45.919 81.109 46.382 82.450 50.261 80.777 50.487 80.96 50.972 79.911 50.765 79.856 52.081 79.094 52214 77.891 53.342 76.953 50.868 77.172 53.302 79.986 53.511 80.670 54.119 79.997 53.921 79.394 55.301 80.832 55.642 80.64 57.2 81.347 57.671 21.322 56.793 82.161 57.422 82.316 51339 22.793 57.6 81.376 56.0e 82.5 56.311 80.428 36.804 79.261 56.897 81.384 56.807 82.326 37.67" 1.019 57M59 22.064 56.156 82207 56.210 83.182 55.60 90173 5.140 80.219 59A0 81.21 59.601 79.787 Ulu4 79.191 61.034 79S22 61688 90552 61.401 20.621 61211 73.144 61.894 76.9M0 -9.909 -9.79 -31.694 -12.180 -12.148 -13.079 -11.207 -9.934 -9.141 -7.566 -6.979 -5.758 -5.487 4.997 -5.228 -4.223 -6.706 -5.521 425 -8.373 -6.973 7.47 -7M58 -8.010 -7.029 -8.020 -5.986 -5.236 .5.918 4.491 -4244 -2.773 -1.815 .0.53 -0.623 -0.092 0.10S 4.979 .7.696 445 48.861 -100335 .33.700 -13,014 -8.610 .7.3w9 -9.3 32 740 -9A95 -10.317 4.91 8 66.26 67.80 0.00 0.00 71.76 0.0 65.90 63.34 O.00 57.35 63.76 65.77 65.21 67.11 0.00 0.00 51.45 46.19 47.07 0.00 42.45 39.63 39.92 35.70 42.39 41.5 43.94 0.00 43.48 47.92 58.49 68.37 73.02 74.14 0.00 0.00 0.00 43.98 40.75 45.99 0.00 47.05 47.39 45.83 S0.59 48.32 49.94 47.32 5347 0.00 57,62 6029 6212 53.99 58310 43 ATOM 56 N ATOM 57 H ATOM 53 CA ATOM 59 Ca ATOM 60 00 ATOM 61 11 ATOM 62 C ATOM 63 0 ATOM 64 N ATOM 65 H ATOM 66 CA ATOM 67 CR ATOM 68 CG ATOM 69 CDI ATOM 70 C12 ATOM 71 C ATOM 72 0 ATOM 73 N ATOM 74 H ATOM 75 CA '9 ATOM 76 CB ATOM 77 CO ATOM 78 CD ATOM 79 QE ATOM g0 oE2 ATOM 81 C ATOM 82 0 ATOM 33 N Inca ATOM 84 H ATOM 85 CA .e ATOM 36 CD ATOM 37 CO ATOM 8 CD ATOM 89 OEI ATOM 90 OE2 ATOM 91 C e ATOM 92 0 44 ATOM 93 N ATOM 94 H AOM 95 CA SAT 96 CE ATOM 97 C ATOM 93 0 ATOM 99 N ATOM too OH ATOM 101 CA ATOM 102 CD ATOM 103 CO ATOM 104 CD ATOM 105 021 ATOM 106 NE2 A7O 07 HE21 ATOM 108 HE22 ATOM 109 C ATOM 110 0 ATOM III N ATOM 112 H ATOM 113 CA ATOM 114 CB ATOM 115 Co ATOM 116 CD ATOM 117 NE ATOM 118 HE ATOM 119 CZ 137 62.538 81.402 137 62.572 81.519 137 63.390 82.210 137 64.149 83,158 137 63.3 33.655 137 62.492 84.111 137 64.313 11.329 137 64.602 80.202 138 64.823 31.79 138 64.686 82.728 138 65.553 80.911 138 65.684 81.695 138 66.536 80.373 138 65.823 79.540 138 66.52 81.749 138 66.813 80.309 138 67.183 79.164 139 67.503 81.099 139 67.248 82.032 139 61.645 80.59 139 69.271 81.757 139 68.277 82.677 139 68.983 93.966 139 68.705 85.009 139 69.811 33.927 139 68.241 79.453 139 68.938 78.458 140 67107 79.556 140 66-57 80.364 140 66.616 78.489 140 65.290 79.74 140 65.411 80.248 140 64.097 80.745 140 63.207 79.947 140 63.971 31.956 140 66.431 77.221 140 66.927 76.166 141 65.703 77298 141 652m6 73.135 141 65.611 76.153 141 64.889 76.570 141 66.979 75.586 141 67.313 74.428 142 67.318 76.487 142 67.532 77.424 142 69.151 76.11S 142 69.866 77.409 142 70.837 77.279 142 70.264 76.716 142 70.722 75.733 142 69.= 77.237 142 0.416 78.075 142 68.852 76.862 142 69.900 75.373 142 70.472 74.312 143 69.911 75.911 143 69.457 76.774 143 70.560 75.235 143 70.398 76.011 143 71.432 77.103 143 71.2% 77.893 143 70.068 73.720 143 69.199 73.300 143 70.151 80.4* -9.833 49.858 -10667 -9.766 -8.792 -9.205 -11.458 .11.086 -12.585 -12.829 .13.478 -14.748 -15.66 -16.097 -17.112 -12.977 -13,115 -12.063 -11.982 -11.330 -10.58 -9.821 -9.426 -10.033 4.510 -10.411 -10.328 -9.711 -9.814 -4.849 -8.234 -7.M -7.015 -6.716 866 -9.621 -9.m7 -10.720 -10.921 -11604 -12.884 -11.947 -11.765 -12.459 -12537 -12.892 -13.316 -i.4S2 -15.714 -16.2386 -16.242 -15.810 -17.056 ,02 -12.010 -10.590 -10.440 -9.482 -1.169 -3.00 4.715 772 -6.81 -6.694 59.34 0.00 62.00 65.68 75.16 0.00 61.26 62.99 61.53 0.00 64.28 63.19 61.50 62.16 65.07 67.11 66.56 68.85 0.00 71.48 73.29 89.52 98.5 99.9 101.79 f9.73 70.27 67.40 0.00 66.30 69.93 79.22 83.51 86.55 89.13 64.12 61.28 64.66 0.00 68.98 10.25 71..4 12.56 72.92 0.00 73.96 77.97 87.44 92.53 95.9 93.80 0.00 0.00 71.44 69.41 70.71 0.00 71.58 66.09 68.24 67.96 68.10 0.00 68.46 d

I

3 i j t l: a 44 ATOM 120 NI ATOM 121 111111 ATOM 122 HH12 ATOM 123 NH2 ATOM 124 1312 ATOM 125 lH2 AMtM 126 C ATOM 127 0 ATOM 128 N ATOM 129 1 ATOM 130 CA ATOM 131 CB ATMI 132 C02 ATOM 133 CoI ATOM 134 CDI ATOM 135 C ATOM 136 0 ATOM 137 N ATOM 138 H 20 ATOM 139 CA ATOM 140 CH A7M 141 CG ATOM 142 CI2 ATOM 144 CE2 ATOM I"4 CE3 ATOM 145 CDI ATOM 146 NEI ATOM 147 HEI ATOM 148 C22 ATOM 149 CM ATOM 150 CH2 ATOM 151 C ATOM 152 0 ATOM 153 N 3O.S ATOM 154 H ATOM 155 CA ATOM 356 CB ATOM 157 C ATOM 158 0 ATOM 159 N ATOM 360 H ATOM 161 CA ATOM 162 CB .ATOM 163 CG ATOM 164 CI ATOM 165 051 ATOM 166 NE2 ATOM 161 H321 ATOM 168 3E22 ATOM 169 C ATOM 170 0 ATOM 171 N ATOM 72 H ATOM 173 CA ATOM 174 Ca ATOM 175 CG ATOM 176 CD ATOMI 177 CE ATOM -178 NZ ATOM 179 HZI ATOM 180 1122 ATOM 181 H3 ATOM 182 -C ATOM 183 0

ARG

ARG

'JO

ARG

ARG

ARG

ARG

ARG

ELE

ELE

ME

MEE

ME

MEE

MEE

ILE

TRP

UPL

TRP

IRP

15

TRP

1RP u-B

TRP

TRP

TRP

TRP

TRP

AUP

TIP

ALA

ALA

AIP

TIP

TIP

TIP

GLN

M.P

GLN

GLN

TLN

GLN

GLN

GLN

GLN

GLN

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

CLYS

143 69.028 80.800 143 68.137 80.358 143 69.090 81.797 143 71.365 80.664 143 72206 80.125 143 17.412 81.661 343 70.013 73.850 143 70.765 72.907 144 68.696 73.662 144 68-1(5 74.411 144 68.143 72.352 144 66.605 72.394 144 66.077 70,990 144 66.084 73.314 144 64.5U 73.540 144 68.604 71.367 144 69.045 70.260 345 68.507 71.765 145 68.167 72.662 145 68.929 70.861 145 68.658 71.462 145 69.038 70.460 145 6117 69.255 145 69.056 68.711 145 67.071 68.594 145 70.133 70.564 145 70.113 69.520 145 70.787 69.359 143 68.642 67562 145 66.717 67.44 145 67.473 66.945 145 70.411 70.581 145 70.817 69.442 146 71.240 71.631 146 70.872 72.546 146 72.678 71.479 146 73.316 72.866 146 73.078 70.678 146 74.075 69.975 147 72.310 70.767 147 71.78 71.421 147 72.543 69.925 141 71.690 70.379 147 72.033 69.642 147 70.829 69.585 147 69.729 70.018 347 70.957 6.991 147 71.821 61.602 147 70.155 68987 147 72.16 68.479 147 73.051 67.595 148 70.969 68.171 141 70.319 6R.869 148 70.621 66.770 148 69.339 66.539 348 69.449 66.861 148 68.211 66.386 348 68.28 66.715 148 69.521 66.190 141 70.339 66.615 141 69.553 65.156 148 69.514 66.411 148 70.457 66.096 148 70.294 64.882 -6,754 4.691 -&.550 -6.498 .6.490 -9.263 -9.074 -9.295 .9.519 .8.995 49.986 -8129 -7.895 4.033 -10.036 -9.743 311.304 -11.514 -12.35L -13.746 -14.839 -15.09% 16136 -14.711 -15,MH .16.475 -17.,366 -16.873 .35.394 -16,445 -12.84 12.042 .12.196 -12279 -12.055 -11.003 -10.3227 -10.833 -9.739 -9.11 -4.583 -7.387 -6090 -5.191 -5.491 -4.ODS -3.760 -3.438 -4.375 4.80 -9.211 -9A39 -9.139 -8.438 -6.954 .6.193 -4.706 4.145 4.627 -4.275 -3.131 4.39.

,10.625 1.00 68.32 1.00 0.00 1.00 0.00 1.00 68.45 1.00 0.00 1.00 00 1.00 73.97 3.00 77.08 1.00 73.91 3.00 0.00 1.0 73.75 3.00 69.05 1.00 69.61 1.00 65.22 1.00 62.44 1.00 72.66 1.00 71.17 1.00 75.70 1.00 0.00 3.00 83.94 3L00 86.15 3.00 93.70 3.00 95.03 1.00 96.30 3.00 94.15 3.00 95.29 1.00 93.47 .OO 0.00 1.00 100.90 3.00 96.37 1.00 98.93 1.00 87.50 1.00 .19 3.00 90.23 .00 0.00 1.00 90.96 1.00 89.47 .OU 92.53 1.00 93.84 1.00 93.24 3.00 0.00 1.00 94.09 1.00 96.74 1.00 102.36 1.00 107.7$ 1.00 109.80 1.00 109.34 3.00 0.00 1.00 0.00 3.00 93.64 1-00 93.43 3.00 91.82 1.00 0.00 1.00 99.79 3.00 93.49 1.00 91.59 3.00 94.76 1.00 97.17 3,00 102316 1,00 0.00 3.00 0.00 .00 0.00 8713 100 88.22 e 45 ATOM 184 N ATOM 185 H ATOM 186 CA ATOM 387 CB S ATOM 188 C ATOM 189 0 ATOM 190 N ATOM 191 H ATOM 192 CA ATOM 193 CB ATOM 194 C ATOM 195 0 ATOM 196 N ATOM 197 H ATOM 198 CA ATOM 199 CB AMM 200 CO ATOM 201 CD ATOM 202 OE ATOM 203 082 ATOM 204 C ATOM 205 0 ATOM 206 N ATOM 207 H ATOM 208 CA ATOM 209 CR ATOM 210 CG2 ATOM 211 CGl ATOM 212 CDI ATOM 213 C ATOM 214 0 ATOM 215 N ATOM 216 H ATOM 217 CA 35 ATOM 218 CR ATOM 219 CO ATOM 220 CDI ATOM 221 CEI ATOM 2n CD2 ATOM 223 CE2 ATOM 224 CZ ATOM 225 OH ATOM 226 10 ATOM 227 C ATOM 228 0 ATOM 29 N ATOM 230 CD ATOM 231 CA ATOM 232 CB ATOM 233 cL ATOM 234 C ATOM 235 0 ATOM 236 N ATOM 237 H ATOM 238 CA ATOM 239 CB ATOM 240 CG2 ATOM 241 CCI ATOM 242 CD, ATOM 243, C ATOM 244 0 ATOM 245 N ATOM 246 HI ATOM 247 CA ALA 149 70.496 66825 ALA 349 70.728 67.775 ALA 149 70273 66.3t6 ALA 149 71.A16 66.848 ALA 349 70.092 4.8017 ALA 149 69.034 64.377 ALA 150 71.079 63.939 ALA 30 71.903 64.290 ALA 150 70.928 62.493 ALA 150 72.254 63.803 ALA 150 69.786 61.803 ALA 350 69.332 60728 (LU 131 69.298 62.391 GLU 151 69.693 6322 GLU 151 68.153 61.823 GLI 151 68.612 61.333 ILU 153 69.510 60.091 GLU 153 69.733 59.292 OLD 153 68.7S3 58.778 OLU 151 70.887 59.159 GLU 15 66.966 62.759 GLU 151 66237 62.811 .E 152 66.751 63-S38 IE I22 67.390 63.5 ILE 152 65S78 64.379 ILE 152 66.006 65.876 ILE 152 64.846 66.776 .E 8 152 66.456 66.419 ME I2 67.026 67.826 E 152 64.88 63.880 0.E 152 65.399 63.521 TYR 353 63.56 63.846 TYR 13 63.125 64.015 T 53 62.606 63.601 TYR 153 61.131 63521 TYh 133 60.815 62.157 TYR 353 60.389 61.23 TYR 353 W0.661 59.769 Tn 153 60.496 62.024 Tn. .53 6066 60.774 TYR 153 60.352 59.65 TP 133 60.116 58.405 TYR 153 59.896 58342 TYR 153 62.734 64.754 TnP 153 62.U5 65.195 PRO 154 63.04S 64.m PRO 154 63.433 63298 PRO 154 63.119 65.885 PRO 354 63.791 65.076 PRO 154 63.408 63.614 PRO 154 63.782 66.309 PRO 154 60.763 65.693 E 35 61.793 67.620 ILE IM5 62.636 68.068 IE I5 60.623 68417 ME 355 60.810 69.687 ILE 355 59.893 70.851 3.E ISS 60.584 69.235 IE 355 59.370 68.340 ME 1 55 60.648 68.636 M.E 355 61.715 68.fi0 IET 156 59.510 68.638 MET 156 58.677 68.455 MET 356 59.471 68.939 -31.691 -11.599 -13.039 -13.906 .13.2TI .13.7m9 -32.604 -13M24 -12.8m2 -12.458 -12.832 -11.366 -11.031 -10.70 -9.316 -9m3 259 -7.7M3 -7.846 -10.610 .63 -11.671 -12.419 -11.820 -11.979 -12.371 -10.633 -10.790 -13063 -14.089 -12.966 -12.080 -14.098 -13.608 -13.006 -13795 -13.266 -11.663 -11.126 -11.934 -11.404 -10.4S1 -15.092 -14.708 -16.332 -16.886 -17.300 -18.508

-IBM

-17.627 -17.860 -17.645 -17.453 -17.912 -17.067 -17.429 -15.624 315.354 -20.136 -19.642 -21542 8.64 0.00 81.28 80.46 60.23 83.20 7.26 0.00 72.54 71.93 67.17 64.24 58.90 0.00 55.45 60.03 67.91 72.08 75.94 70.44 49.03 40.75 46.24 0.00 41.23 41.97 3744 39.50 45.38 39.42 38.70 37.18 0.00 34.02 30.99 28.21 27.24 25.65 24.92 20.91 24.92 22.74 0.00 31.41 30.52 29.09 2614 26.59 21.73 26.66 27.17 30.28 27A9 000 27.05 27.3 28.62 27.98 30.26 33.41 34.72 33.95 0.00 31.70 2 r: g /r 46 ATOM 248 CR ATOM 249 CG AT(Al 250 SD ATOM 251 CE ATOM 252 C ATOM 253 0 ATOM 254 N ATOM 255 H ATOM 256 CA AOMM 257 Ca ATOM 258 CG ATOM 259 ODI ATOM 260 OD2 ATOM 261 C ATOM 262 0 ATOM 263 N ATOM 264 R ATOM 265 CA ATOM 26 CB ATOM 267 CG ATOM 268 CD ATOM 269 CE ATOM 271] N! ATOM 271 HZI ATOM 272 1Z2 ATOM 273 un ATOM 274 C ATOM 275 0 ATOM 276 N ATOM 277 H ATOM 278 CA ATOM 279 CE ATOM 280 o ATOM 28 H 35 ATOM 22 C ATOM 283 0 ATOM 284 N ATOM 285 H ATOM 286 CA ATOM 287 CH ATOM 288 00 ATOM 289 HG ATOM 290 C *ATOM 291 0 45 ATM 292 N ATOM 293 H ATOM 294 cA ATOM 295 CE ATOM 296 CG ATOM 297 CD ATOM 298 NE ATOM 299 HE ATOM 300 c2 ATOM 301 NH3 ATOM 302 111113 ATOM 303 31112 ATOM 30 NH2 ADtO 305 I1H21 ATOM 306 H22 ATOM 307 C ATOM 308 0 ATOM 309 N ATOM 310 H AO. 311 CA MET 156 MET 1356 MET 156 MET 156 ME 156 MET 156 ASP 157 ASP 157 ASP 157 AS? I57 ASP 157 ASP 157 ASP 157 ASP 157 ASP I5 LYS 158 LYS 158 LYS 158 LYES 58 LYS 158 LY 158 LYS 158 LYS 158 LYS 158 LYS 358 LYS 358 LYS 158 LYS 158 SER 159 SER 159 SER 159 SIR 159 SER 159 SER 159 SER 359 SER 159 SER 160 SIR 160 SIR 160 SER 160 SER 160 SIR 160 SER 160 SER 160 AILG 161 ARG 163 ARG 161 MAG 161 ARG 161 AG 161 ARG 151 ARO 161 MRG 161 ARO 161 ARC 161 ARG 161 eato 161 ARG 161 ARG 16i ARG 161 A R C 1 6 1 THR 162 TUR 162 THR !62 58.252 68.339 -22M4 59,574 67.001 -22.838 57.138 66.=5 -237 57.590 64.545 -23M8 5934 70.434 -21.735 59.035 71.208 .20.05 59.711 70.869 -22.942 59.923 70.224 -23.648 59.715 72-281 -23240 60-385 72.38 -24.613 60.369 73.848 -25.178 60.483 73.957 -26.401 60.249 74.822 -24.421 58.315 72.829 -23.20- 57.360 72.284 -23730 58.22 73.965 -22.543 59.032 74-318 -22.102 16.975 74.38 -22.320 37.265 75.906 -21540 56.037 76.664 -21.085 56.390 71.826 -20.34 55.152 78.472 -19.54 5537 79.342 -18.433 56.154 80095 -38.79 56.050 78.792 -17.712 34691 79.763 -17.998 "6.275 74.942 -23.618 55.076 74.80 -23.735 56.962 75.367 -24.672 57933 75.470 -24.645 56297 75.717 -25-916 57.3 7622 26.930 58553 75.55 -26.729 58.453 74.606 -26.880 55.543 74-M9 -26548 54.666 74.774 -70 55.886 73.373 -26.184 56.444 73.181 -25399 55.410 72.61 -26.959 56592 71.756 -2.769 .835 71.958 -27.083 58.517 71.3 -27.614 54.3m 713154 -26121 5.2 70317 -6.00 4.82 71.162 -24.782 55.201 71.923 -24.278 S4.322 70.004 -24.097 54.987 69.929 -22.727 54.633 71.054 -23.769 55-s9 71.078 -20.57! 55.148 72.182 -19.760 54.189 72.357 -19.653 56.001 72.971 -19.138 55.486 74.036 -18.4 54.499 74.198 -1&481 56.093 74.673 -18.006 57.339 72.712 -19.11S 57.703 71.901 -39.51 57.957 73.33 1 -18.630 52.816 70.07 -23.999 52.219 71.144 -23.983 52.134 68.955 -23.937 W295 68.099 -24.073 50699 68.963 -23.791 3.00 1.00 1.00 3oo .00 1.00 1.00 .00 .00 3.00 1.00 .oo 1.00 3.00 1.00 1.00 Looo i.00 3.00 1.0 1.00 1.00 Loo 1.00 Loo 1.00 1.00 1.00 3.00 1.00 1.00 Looo 3.00 1.00 3.00 1.00 1.00 3.00 1.00 3.00 3.00 1.00 3.00 1.00 3.00 1.00 1.00 1.00 3.00 3.00 1.00 1-00 .30 1.00 1.00 1.00 .00 31.0 30.35 39.82 34.89 34.78 32.33 37.95 0.00 38.36 51.68 61.42 68.30 67.34 31.15 2750 30.63 0.00 33.69 34.01 40.80 48.58 52.62 59.47 0.00 0.00 0.00 38.44 4.75 41.20 0.00 39.99 39.69 40.47 0.00 40.83 43.09 43.73 0.00 4571 45.15 53.30 0.00 45.78 48.03 38.45 0,00 32.89 31.71 29.23 26.52 23.86 0.00 24.88 23.22 0.00 0.0 26,60 0.00 0.00 30,40 30.63 25.20 0.00 23,14 i

I

i i ;t .or 47 ATOM 312 CB lER 16 50.068 68176 -25.028 1.00 23.65 ATOM 313 001 Ml 162 67.056 1.00 2656 ATOM 314 HGI THR 162 50.21 66510 -24.445 1.00 0.00 ATOM 315 C02 71 162 .,149 69.114 -26.306 1.00 23.19 ATOM 316 C 1111 162 30.373 68219 -22.510 1.00 25.37 ATOM 317 0 THR 162 50.062 67.030 -22. 1.00 27.7S ATOM 318 N ARG 163 .423 68.26 -21.340 1.00 4.62 ATOM 319 R ARG 163 X50. 69.79 -21.312 1.00 000 ATOM 320 CA AG 163 50.174 68.133 -20.077 1.00 24.12 ATOM 321 CH ARG 163 50.757 68.905 -18.918 1.00 15.46 ATOM 322 CO ARO 163 52.227 68.612 -18.793 1,00 17.34 ATOM 323 CD ARG 163 52.721 69.593 -17.772 1.VD 18.83 ATOM 324 NE ARG 163 514. m 69.427 -17.669 1U Z4.11 ATOM 325 HE ARG 163 54.5V. 68.736 -13.195 1.00 0.00 ATOM 326 CZ ARO 163 4.22 70.22 -16883 1.00 25.67 ATOM 327 HIl ARO 163 56.160 70.051 -16.807 1.00 34.46 ATOM 328 1111 ARG 163 56.596 69318 -17328 1.00 0.00 ATOM 329 HHIZ ARG 163 56.710 70.634 -16209 1.00 0.00 ATOM 330 tN1U ARG 163 54.219 7120 -16.167 1.D0 27.3 ATOM 331 H21 ARG 163 5329 71357 -16223 1.00 0.00 ATOM 332 Hl2 ARO 163 54.772 71.806 -15.57S L. 0.00 ATOM 333 C ARO 163 49.738 67.876 -19.714 1.00 24.62 ATOM 334 0 ARO 163 41.893 68.762 -19.750 1.00 29.04 ATOM 335 N LEU 164 49,357 66667 -19.30 1.00 23.75 ATOM 336 H LEO 164 49.023 65.971 .19.147 1.00 0.00 AT01 337 CA LEU 164 46.944 6&55 -19.043 1.03 24.S, ATOM M CB LEU 164 46.3M6 65-3 -19.893 1.00 27.72 ATOM 339 CO LEO 164 4.556 65.607 -21.165 1.00 28.04 ATOM 340 CDI LEO 164 46.108 66.89 -21.926 1.00 30.94 ATOM 341 Cn LE .164 45.73 6.331 2LO6 1.00 27.16 ATOM 342 C LEO 164 46.774 66.041 -17.612 100 23.76 ATOM 343 0 LEU 164 47.607 65.28 -17.142 1.00 22.71 ATOM 344 1 AL 165 45.733 66505 -16907 1.00 21.51 ATOM 345 H ALA 165 45.124 61.175 -17.22 .00 0.00 35 ATOM 346 CA ALA 165 45.396 65.986 -1599 1.00 24.14 ATOM 347 CE ALA 16S 45.68 67.006 -14.4 1.00 21.16 ATOM 343 C ALA 165 43.924 65.6 -153 1.00 24.40 ATOM 349 0 ALA 165 43.162 66279 46279 1.00 25.81 ATOM 350 1 LEU 166 43.495 64.730 -14699 1.60 24.86 ATOM 351 H LEO 166 44.143 64.82 .14.114 .OD 0.00 ATOM 352 CA LEO 166 42.091 64.361 .14.592 1.00 27.92 ATOM 353 CE LEO 166 41.914 62.948 -15.196 1.00 23.03 ATOM 354 CG LEO 166 40.511 62.210 -14.919 1.00 20.45 ATOM 355 CDI LED 166 39.446 62.990 -15.65 1.00 14.43 45 ATOM 356 CD2 LEO 166 40.691 60.07 -15.320 1.00 21.71 ATOM 357 C LEO 166 4150 64,404 -13.149 1.CO 23.64 ATOM 338 0 LEU 166 42.207 63.33 -12.61 1.00 26.66 ATOM 339, N ME 167 40.441 65.079 -12.901 1.00 27.07 ATOM 360 H a.E 161 39.991 63.549 .13.638 1.00 0.00 ATOM 361 CA ILE 167 39.802 65.137 -11.600 1.00 2.8 ATOM 362 CEB LE 16? 39.494 662 -11.20 1.00 20.90 -ATOM 363 CG2 MEl 167 33.719 66.663 -9.893 1.00 21.77 ATOM 364 CI IL 167 40.193 67-354 -11.003 1.00 17.47 ATOM 365 CDI MlE 167 40.499 63.753 -10.522 1.00 17.98 ATOM 366 C ME 167 3.513 64.35 -11.634 1.00 24.82 ATOMA 367 0 ILE 167 37.634 64.653 -12.493 1.00 23.12 ATOM 368 N IL 168 38333 63321 -10.865 1.00 25.26 ATOM 369 H IL 168 39.066 63.004 -1029 1.00 0.00 ATOM 3170 CA IE 168 37.022- 62.731 -10.751 1.00 23.44 ATOM 31 C ILE 168 37.119 61.228 -11.016 1.0 23.97 ATOM 372 CG2 ILE 168 35.741 6032 10.6 1.00 28.90 ATOM 373 CI 12. 163 37531 61.030 -12.5W6 1.00 22.39 ATOM 374 CDI am 168 37.869 39.! -12.959 1.00 22.21 ATOM 375 C ILE 168 36.0 62.931 -9353 1.00 24.34 48 ATOM 376 0 ATOM 37 1 ATOM 373 H ATOM 379 CA ATOM 380 CB ATOM 381 So ATOM 332 C ATOMf 393 0 ATOM 3u4 N ATOM 35 H ATm 336 CA ATOM 337 CB ATOM 393 CG ATOM 339 ODI ATOM 390 N 2 ATOM 391 H4021 ATMO 392 1122 ATOM 393 C ATOM 0 20 ATOM 395 N ATOM 396 H ATOM 397 C *ATOM 398 CE ATOM 399 CG ATOM 40 C) ATOM 401 022 ATOM 402 OE2 ATOM 403 C ATOM 404 0 ATOM 40 N ATOM 406 H ATOM 407 CA ATOM 408 CB ATOM 409 CO ATOM 410 CD ATOM 411 022 ATOM 412 032 ATOM 413 C ATOM 414 0 40 ATOM 41S 1 ATOM 416 H ATOM 417 CA ATOM 418 Ca *AVOM 419 CG ATOM 42 CD *ATOM 42!1 CM, ATOM 422 CEI ATOM 413 CE2 ATOM 424 CZ ATOM 425 C ATOM 426 0 ATOM 4-7 N ATOM 428 1 ATOM 4-9 CA ATOM 430 CE ATOM 431 CO ATOM 432 ODI ATOM 433 002 ATOM 434 C ATOM 435 0 ATOMt 436 1 ATOM 437 H ATOM A3U1 CA ATOMt _439.- CB

LE

CYS

CYS

CYS

CYS

CYS

CYS

CYS

ASN

ASN

ASN

ASN

ASN

ASN

GLU

GLV

CLD

GLV

GLU

OLD

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GL GLU

GLU

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GLU

PIE

PHE

PHE

PIlE

M~E

PIlE

PRE

ME

PRE

PRlE PlE

APH

ASP

ASP

ASP

ASP

ASP

A'SP

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ASP

SER

SEP.

SER

SEP.

68 37.1.6 62.633 169 35.337 63M613 169 34.821 63.777 169 34.765 64.030 169 34.347 65.54 169 34.147 66.177 169 33.323 63.629 169 32520 63.950 ItO 32.9M3 62.918 170 33.617 62.U3 170 31.554 62.555 170 31.3M. 61.07! 170 31674 60.138 170 31.709 60.485 170 31.7m 5.87 170 32.015 58.569 170 32.132 53.288 170 30.957 63.328 17 31.382 63.066 M.1- 30.000 64.247 171 29.711 64.420 171 29.301 6.05 171 29.119 66.413 171 28.418 67.094 171 28.233 63579 171 28302 69.095 171 28003 69.233 171 27.914 64.338 171 27S09 64.214 172 27.133 63.976 172 27505 63.929 172 25.760 63534 172 24.802 64.170 172 23.819 65.134 172 24.170 66.50 172 23149 67.402 172 25.42 66843 172 25.676 62.035 172 2.92 61.51 173 25.125 61.338 173 24.495 61.780 173 250S5 59.833 173 25.178 59.313 173 27.371 59.57 173 2&173 ra.5V1 173. 27.927 60.747 173 29-535 58.684 173 29191 60.94 173 30.089 59.70 373 23.661 59372 373 22.772 60.010 374 23.394 58.194 174 24.131 57.657 174 2.544 57.63 174 21.943 56-370 174 22.091 56.666 174 22.150 55.699 174 22.148 57.&M 174 21.500 57.347 174 20.317 57447 173 22.354 56940 175 .23.312 57.026 175 21.856 56&461 I 22.007 54.933 -9.356 .10.079 -7906 -7.831 6.282 -7.866 735 6803 -&124 -6.619 -4286 -7-440 _&614 -7351 216 901 -5-478 -4.361 -5.672 -4.822 .3*634 -3.17 -4.M9 4-761 -4.240 -3.096 -5251 43.050

_&M

-5.4m8 -3.699 -3.708 -3.911 -5242 -266 -4.250 -3,493 -4303 -3.142 -3286 -3.76R -2.867 -3,773 -2.902 -3332 -4.221 -3.675 -4.764 .5.134 4.745 .5.557 -7030 I.795 -7.408 -3.371 -3.03 -2.457 -2.628 -1.185 -I.132

.OG

1.00 1.00 1.00 1.0 .00 .00 1,00 1.00 1.00 .00 1.00 1.00 .oo 1.00 1.00 1.00 .00 .00 1.0 IMa 1.00 .00 .oo 1.00 1.00

LOD

1.00 .oo 1.00 .00 L~oo

LOD

1.00 .00 .00 Loua 1.00 .00 .00 .00 .00 1.00 .00 1.00 1.00 .00 1.00 1.00 1.00 100 1.00 1.00 1.00 1.00 1.00 3.00 23.09 26,60 0.00 27.2 26.82 31.50 28.36 25.3 27.23 0.00 29.76 26.22 29.16 32.47 25.4 0.00 0.00 32.01 M4.71 36.94 0.00 39.76 36,97 43.95 47.95 53.49 48.17 41.90 40.9 43.24 0.00 51.31 55.01 66.24 74.35 31.11 76.04 49.73 46.42 49.39 0.00 53,47 50.77 46.81 40.36 42.17 37.00 33.90 39.65 56.41 00.33 54.59 0.00 53.81 51.10 53.87 53.10 53.80 56.79 60.64 57 04 0.00 5%.32 53.73

A

48 ATOM 376 0 ATOM 377 N ATOM 378 H ATOM 379 CA ATOM 380 CB ATOM 381 SG ATOM 382 C ATOM 383 0 ATOM 384 N ATOM 395 H ATOM 386 CA ATOM 387 CB ATOM 388 CG ATOM 389 ODI ATOM 390 ND2 ATOM 391 HD21 S ATOM 392 HD22 SATOM 393 C S ATOM 394 0 S 20 ATOM 395 N ATOM 396 H S ATOM 397 CA SATOM 398 CB SATOM 399 CG ATOM 400 CD ATOM 401 OEI ATOM 402 OE2 ATOM 403 C ATOM 404 0 ATOM 405 N ATOM 406 H ATOM 407 CA S ATOM 408 CB ATOM 409 CG 35 ATOM 410 CD S ATOM 411 OEI ATOM 412 OE2 ATOM 413 C S ATOM 414 0 40 ATOM 415 N ATOM 416 H ATOM 417 CA ATOM 418 C3 ATOM 419 CG ATOM 420 CDI ATOM 421! CD2 ATOM 422 CEI ATOM 423 CE2 ATOM 424 CZ ATOM 425 C ATOM 426 0 ATOM 427 N ATOM 428 H ATOM 429 CA ATOM 430 CB ATOM 431 CG ATOM 432 ODI ATOM 433 OD2 ATOM 434 C ATOM 435 0 ATOM 436 N ATOM 437 H ATOM 438 CA ATOM 439 CB

ICE

CYS

CYS

CYS

CYS

CYS

CYS

CYS

ASN

ASN

ASN

ASN

ASN

ASN

ASH

ASN

ASN

ASN

ASN

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

GLU

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PIlE

PIE

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PRE

PHE

PHE

PHE

PHE

PHE

PHE

ASP

ASP

ASP

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ASP

ASP

ASP

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ASP

SER

SER

SEX

SER

168 37.126 62.633 169 35.337 63.613 169 34.821 63.777 169 34.765 64.030 169 34.847 65.544 169 34.147 66.177 169 33.323 63.629 169 32.520 63.950 170 32.948 62.918 170 33.617 62.683 170 31.554 62.555 170 31.372 61.071 170 31.674 60.138 170 31.709 60.485 170 31.970 58.887 170 32.015 58.569 170 32.132 58.288 170 30.957 63.328 170 31.382 63.086 171 30.000 64.247 171 29.711 64.420 171 29.301 64.905 171 29.119 66.413 171 2S.418 67.084 171 28.233 68.579 171 28.302 69.095 171 28.003 69.233 171 27.914 64.338 171 27.509 64.214 172 27.133 63.976 172 27.505 63.929 172 25.760 63.534 172 24.802 64.170 172 23.819 65.134 172 24.170 66.580 172 23.249 67.402 172 25.342 66.893 172 25.676 62.035 172 26.092 61.501 173 25.125 61.338 173 24.695 61.780 173 25.085 59.888 173 25.878 53.313 173 27.371 59.547 173 28.173 58.521 173 27.927 60.747 173 29.535 58.684 173 29.291 60.904 173 30.089 59.870 173 23.661 59.372 173 22.772 60.010 174 23,394 58.194 174 24.131 57.657 174 22.044 57.658 174 21.943 56.370 174 22.091 56.666 174 22.150 55.698 174 22.148 57.850 174 21.500 57.347 174 20.317 57.447 175 22.354 56.940 175 23.312 57.026 175 21.8S6 56.461 175 22.007 54.938 -8.356 -9.260 -10.079 -7.986 -7.831 .6.282 -7.866 -8.735 46.803 .6.124 -6.619 -6.286 .7.440 .8.614 -7.151 -6.216 .7.901 -5.478 -4.361 -5.672 -6.594 -4.558 4.822 -3.634 -3.777 -4.890 -2.761 -4.240 -3.096 -5.251 -6.159 -5.050 .6061 -5.408 -5.699 -5.708 -5.911 -5.242 -6.266 4.250 -3.485 4.303 -3.142 -3.286 -3.768 -2.867 -3.778 -2.902 -3.332 -4.221 -3.675 ,4.764 -5.134 -4.745 -5.557 -7.030 -7.795 -7.408 -3.370 -3.086 -2.457 -2.628 -1.185 -1.152 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 23.09 26.60 0.00 27.82 26.82 31.50 28.36 25.83 27.23 0.00 29.76 26.22 29.16 32.47 I. .J) 0.00 32.01 34.71 36.94 0.00 39.76 36.97 43.98 47.95 53.49 48.17 41.90 40.97 48.24 0.00 51.31 55.01 66.24 74.35 81.11 76.04 49,78 46.42 49.39 0.00 53.47 50.77 46.81 40.36 42.17 37.00 38.90 39.65 56.41 60.38 54.59 0.00 53.81 51.10 53.87 53.10 53.80 56.79 60.64 57.04 0.00 56.82 523.73 n.j 4'- 49 ATOM 440 OG SER ATOM 441 HG SER ATOM 442 C SER ATOM 443 0 SER ATOM 444 N ILE ATOM 445 H ILE ATOM 446 CA ILE ATOM 447 CB ILE ATOM 448 CG2 I.E ATOM 449 CG1 ILE ATOM 450 CDI IE ATOM 451 C ILE ATOM 452 0 ILE ATOM 453 N PRO ATOM 454 CD PRO ATOM 455 CA PRO ATOM 456 CB PRO ATOM 457 CG PRO S ATOM 458 C PRO 20 ATOM 459 0 PRO ATOM 460 N ARG S ATOM 461 H ARG ATOM 462 CA ARG S ATOM 463 CB ARG ATOM 464 CO ARG S ATOM 465 CD ARG S ATOM 466 NE ARG ATOM 467 HE ARG ATOM 468 CZ ARG ATOM 469 NHI ARG ATOM 470 HHII ARG ATOM 471 HH12 ARG ATOM 472 NH2 ARG ATOM 473 HH2I ARG 35 ATOM 474 HH22 ARG S ATOM 475 C ARG ATOM 476 0 ARG ATOM 477 N ARG ATOM 478 H ARG ATOM 479 CA ARG SATOM 480 CB ARG ATOM 481 CG ARG ATOM 482 CD ARG SATOM 483 NE _ARG ATOM 484 HE ARG ATOM 485 CZ ARG ATOM 486 NHI ARG ATOM 487 HHI I ARG ATOM 488 HH12 ARG ATOM 489 NH2 ARG ATOM 490 HH21 ARG ATOM 491 HH22 ARG ATOM 492 C ARG ATOM 493 0 ARG ATOM 494 N THR ATOM 495 H THR SATOM 496 CA THR ATOM 497 CB THR ATOM 498 OGI THR ATOM 499 HGi THR ATOM 500 CG2 THR ATOM 501 C THR ATOM 502 0 THR ATOM 503 N GLY 175 21.821 54.307 175 22.453 54.685 175 22.614 57,115 175 22.050 57.584 176 23.949 57.172 176 24.384 56.889 176 24.725 57.758 176 25.167 57.155 176 25.982 55.645 176 26.947 57.381 176 28.205 56.486 176 24.716 59.262 176 24.516 59.755 177 24.918 60.009 177 25.067 59.502 177 24.821 61.475 177 24.733 61.862 177 25.513 60.744 177 25.870 62.308 177 27.078 62.045 178 25.287 63.373 178 24.351 63.552 178 25.959 64.288 178 24.910 65.270 178 25.505 66.396 178 24.484 67.300 178 25.167 68.507 178 26.144 68.522 178 24.473 69.607 178 23.102 69.585 178 22.609 68.736 178 22.586 70.415 178 25.133 70.773 178 26.133 70.802 178 24.609 71.605 178 27.133 65.020 178 27.078 65.589 179 28.243 65.007 179 28.218 64.607 179 29.458 65.625 179 30.625 64.919 179 30.599 63.395 179 31.759 62.682 179 33.057 63.164 179 33.387 62.869 179 33.827 63.996 179 33.483 64.391 179 32.630 64.073 179 34.077 65.023 179 34.965 64.483 179 35.205 64.214 179 35.562 65.100 179 29.512 67.124 179 30.215 67.684 180 28.730 67.862 180o 28.063 67.420 180 28.816 69.308 180 27.770 69.840 180 26.517 69.456 180 25.800 69.689 180 27.792 71.351 180 30.221 69.783 180 30.882 69.285 181 30.713 70.783 -2.423 -3.041 -0.038 0.945 -0.133 -0.963 0.942 0.920 1.127 -0.375 -0.451 0.770 -0.332 1.801 3.162 1.806 3.280 3.937 1.096 1.042 0.539 0.778 .0.328 -0.842 -1.661 .2.273 -2.664 -2.727 -2.918 .2.899 -2.708 .3.103 -3.165 -3.145 -3.354 0.2f0 1.354 -0.439 -1.336 0.039 .0.632 -0.447 -1.159 .0.704 0.169 .1.427 .2.691 .3.103 .3.191 .0.859 0.074 -1.370 .0.198 1.034 0.570 1.138 0.548 1.532 0.968 1.56S 1.735 0.901 1.807 0.171 50.93 0.00 59.19 63.80 57.13 0.00 50.84 52.16 54.09 51,82 47.41 47.11 45.21 47.09 46.36 48.52 45.72 45.89 51.58 56.38 50.95 0.00 52.24 54.80 62.55 67.32 77.79 0.00 85.85 92.67 0.00 0.00 90.17 0.00 0.00 49.82 51.03 47.03 0.00 42.75 42.27 39.14 40.61 40.03 000 40.60 35.23 0.00 0.00 35.40 0.00 0.00 44.55 44.42 45.86 0.00 46.36 50.60 54.63 0.00 53.13 4.6.41 47.93 46.30

I

I

50 ATOM 504 H GLY ATOM 505 CA GLY ATOM 506 C GLY ATOM 507 0 GLY ATOM 508 N ALA ATOM 509 H ALA ATOM 510 CA ALA ATOM 511 CB ALA ATOM 512 C ALA ATOM 513 0 ALA ATOM 514 N GLU ATOM 515 H GLU ATOM 516 CA GLU ATOM 517 CB GLU ATOM 518 CG GLU ATOM 519 CD GLU ATOM 520 OEI GLU ATOM 521 OE2 GLU ATOM 522 C GLU ATOM 523 0 GLU ATOM 524 N VAL ATOM 525 H VAL ATOM 526 CA VAL S ATOM 527 CB VAL ATOM 528 CGI VAL ATOM 529 CG2 VAL ATOM 530 C VAL ATOM 531 0 VAL ATOM 532 N ASP ATOM 533 H ASP ATOM 534 CA ASP ATOM 535 CB ASP ATOM 536 CO ASP SATOM 537 ODI1 ASP 35 ATOM 538 OD2 ASP ATOM 539 C ASP ATOM 540 0 ASP ATOM 541 N ILE SATOM 542 H ILE ATOM 543 CA ILE ATOM 544 CB ILE ATOM =545 CG2 IE ATOM 546 CGI ILE ATOM 547 CDI ILE ATOM 54 C ILE ATOM 549 0 ILE ATOM 550 N THR ATOM 551 H THR ATOM 552 CA THR ATOM 553 CB THR ATOM 554 OGI THR ATOM 555 HGI THR ATOM 556 CG2 THR ATOM 557 C THR ATOM 558 0 THR ATOM 559 N GLY ATOM 560 H GLY ATOM 561 CA GLY ATOM 562 C GLY "ATOM 563 0 GLY ATOM 564 N MET SATOM 565 H MET SATOM 566 CA MET ATOM 567 CB MET 181 30.090 71.312 181 32.118 71.193 181 32.915 70.679 181 34.005 71,154 182 32.381 69.688 182 31.540 69.257 182 33.136 69.171 182 32.314 68.028 182 33.503 70.220 182 34.590 70.239 183 32.599 71.153 183 31.748 71.149 183 32.G24 72.180 183 31.614 73.091 183 31.637 73.914 183 30.918 73.216 183 30.272 73.920 183 30.994 71.988 183 33.998 73.045 183 34.778 73.390 184 34.197 73.424 184 33.502 73.236 184 35.437 74.093 184 35.391 74.286 184 36.568 75.090 184 34.081 74.992 184 36.707 73.335 184 37.716 73.836 185 36.649 72.025 185 35.811 71.642 185 37.768 71.151 185 37.470 69.728 185 37.375 69.592 185 38.047 70.367 185 36.635 68.705 185 38.020 71.139 185 39.16 71.233 186 36.956 71.032 186 36.070 70.902 186 37.096 71.078 186 35.728 70.861 186 35.874 71.027 186 35.212 69.446 186 33.829 69.127 186 37.684 72.413 186 38.691 72.466 187 37.132 73.560 187 36.310 73.583 187 37.739 74.320 187 36.943 75.985 187 35.600 75.803 187 35.578 75.839 187 37.461 77.371 187 39.193 74.935 187 39.997 75.448 188 39.561 74.456 188 38.908 74.010 188 40.932 74.607 188 41.872 73.681 188 42.983 74,024 189 41.491 72.438 189 40.618 72.115 189 42.405 71.582 189 42.016 70.117 -0.350 0.236 .0,948 -1.278 -1.672 -1.418 -2.785 -3.377 -3.824 -4,378 -4.107 -3.627 -5.108 -5,246 -6.544 -7.697 -8.4S1 .7.818 -4,776 -5.639 -3.521 -2.857 -3,114 -1.598 -1.054 -1.308 -3.505 -3.987 .3.278 -2.945 .3.586 -3.077 -1,539 .0,833 -1.082 .5.059 -5.493 -5,849 -5.447 -7.305 4-8.016 .9.544 -7.688 -8.292 -7.748 -8.447 -7,388 -6.842 -7.788 -7.175 -7.625 -3.583 -7.566 -7.395 -8.163 -6.203 -5.619 -5.753 -6.483 -6.B71 -6.720 -6.415 -7.426 -7.218 0.00 41.25 40.64 37.29 35.93 0.00 39.67 35.68 39.67 40.75 42.72 0.00 40.12 47.30 63.74 71.48 78.62 76.00 35.44 37.02 32.69 0.00 32.56 30.19 28.08 34.05 31.95 31.94 36.44 0.00 32.53 33.96 35.35 28.61 32.20 30.28 29.51 29.28 0,00 31.19 31.31 29.98 33.52 24.25 30.16 30.56 27.47 0.00 27.93 27.45 32.61 0.00 24.19 28.16 28.74 28.71 0.00 25.69 27.48 28,6i 27.34 0.00 25.62 31.58 .1 51 ATOM 568 CG ATOM 569 SD ATOM 570 CE ATOM 571 C ATOM 572 O0 ATOM 573 N ATOM 574 H ATOM 575 CA ATOM 576 C ATOM 577 0GI ATOM 578 HG ATOM 579 CG2 ATOM 580 C ATOM 581 O0 ATOM 582 N ATOM 583 H S ATOM 584 CA ATOM 585 CB ATOM 586 CGo ATOM 587 SD S ATOM 588 CE S ATOM 589 C ATOM 590 O0 ATOM 591 N ATOM 592 H ATOM 593 CA ATOM 594 CB ATOM 595 CG ATOM 596 C9D! ATOM 597 CD2 ATOM 598 C ATOM 599 O0 ATOM 600 N ATOM r01 H S ATOM /502 CA S603 CD ATOM 604 CG ATOM 605 CDI ATOM 606 CD2 S 40 ATOM 607 C ATOM 608 O0 ATOM 609 N ATOM 610 H ATOM 611 CA ATOM 612 C3 ATOM 613 CG ATOM 614 CD ATOM 615 OEI ATOM 616 NE2 ATOM 617 HE21 ATOM 618 HE22 ATOM 619 C ATOM 620 0 ATOM 621 N ATOM 022 H ATOM 623 CA ATOM 624 CB ATOM 625 CG ATOM 626 ODI ATOM 627 ND2 ATOM 628 HD21 ATOM 629 HD22 ATOM 630 C ATOM 631 0O

MET

MET

MET

MET

MET

THR

THR

THR

THR

HR

THR

THR

THR

MFIR MET MET ME

MET

MET

MET

MET

MET

MET

LEU

LEU

LEU

LEU

MEU

MEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

GLN

GLN

GLN

GLN

GLN

GLN

GLN

GLN

GLN

GLN

GLN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

189 42.445 69.542 -5.843 189 44.201 69.754 -5.409 189 44.983 68.533 6.417 189 42.477 71.893 -8.893 189 43.559 71.814 -9.460 190 41.385 72.251 -9.570 190 40.494 72.204 -9.158 190 41.446 72.677 -10.959 190 40.030 73.023 .11.453 190 39.259 71.861 -11.228 190 38.355 72.025 -11.500 190 39.922 73.321 -12.943 190 42.363 73.878 -11.143 190 43.255 73.913 -11.989 191 42.207 74.935 -10.353 191 41.531 74.956 -9.640 191 43.092 76.058 .10.563 191 42.618 77.263 -9.731 191 41.203 77.811 -10.021 191 40.720 77.816 -11.767 191 42.005 78.898 -12.322 191 44.534 75.727 -10.227 191 45.439 76.102 -10.949 192 44.841 75.015 -9.139 192 44.141 74.785 -8.487 192 46.207 74.599 872 192 46.226 73.695 -7.637 192 47.592 73.125 -7.246 192 48.470 74.276 -6.797 192 47.450 72.049 -6.153 f92 46.798 73.872 -10.049 192 47.871 74.187 -10.546 93 46.115 72.857 -10.554 193 45.208 72.654 -10.237 193 46.743 72.049 -11.571 193 45.920 70.802 -11.849 193 46.081 69.749 -10.749 193 45.125 68.587 -11.000 193 47.509 69.236 -10.751 193 46.921 72.815 -12.844 193 47.962 72.768 -13.484 194 45.909 73.565 -13.262 194 45.038 73.515 -12.809 194 46.071 74.455 -14.391 194 44.769 75.244 -14.530 A; 44.400 75.460 -15.983 194 42.994 75.954 -16.101 194 42.085 75.270 -16.546 194 42.733 77.202 -15.779 194 43.468 77.783 -15.495 194 41.799 77.487 -15.61 194 47.281 75.347 .14-146 194 48.177 75.481 -14.964 195 47.356 75.986 -12.990 195 46.613 75.928 -12.354 195 48.530 76.762 -12.606 195 48.460 77.239 -11.151 195 47.691 78.525 -10.957 195 47.679 79.452 -11.753 195 47.015 78.668 -9.836 195 47.044 77.925 -9.206 195 46.506 79.497 -9.715 195 49.860 76.066 -12.719 195 50.871 76.696 -12.949 1.00 1.00 1,00 1.o00 1.00 1.00 1.00 1.00 1.00 1.00 .00 .00 .00 .00 .00 1.00 .00 1.00 1.00 1.00 I.00 1.00 I.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 I.00 1.00 1.00 I.00 1.00 1.00 I.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 Loc) 1.00 2.00 1.00 1.00 I.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 37.74 37.50 38.22 24.18 27.99 24.30 0.00 23.16 23.46 23.54 0.00 19.04 25.40 25.28 25.85 0.00 26.80 31.82 30.18 42.05 40.6 26.94 28.00 25.81 0.00 22.18 19.29 25.33 25.29 25.35 19.90 19.35 10.99 0.00 21.52 21.31 23.69 26.09 24.27 25.18 25.97 27.74 0.00 29.82 28.29 33.28 34.79 33.62 36.58 0.00 0.00 31.33 31.54 33.01 0.00 30.77 34.88 38.35 43.57 36.28 0.00 0.00 28.51 26.10 1~ 52 ATOM 632 N ATOM 633 H ATOM 634 CA ATOM 635 CB ATOM 636 CG ATOM 637 CDI ATOM 638 CD2 ATOM 639 C ATOM 640 0 ATOM 641 N ATOM 642 H ATOM 643 CA' ATOM 644 C ATOM 645 0 ATOM 646 N ATOM 647 H eATOM 648 CA ATOM 649 CB ATOM 650 CG 20 ATOM 651 CDI ATOM 652 CEI ATOM 653 CD2 ATOM 654 CE2 S ATOM 655 CZ S ATOM 656 OH ATOM 657 HH ATOM 658 C ATOM 659 0 ATOM 660 N ATOM 661 H ATOM 662 CA ATOM 663 CB ATOM 664 OG3 ATOM 665 IIG 35 ATOM 666 C ATOM 667 0 ATOM 668 N ATOM 669 H ATOM 670 CA S" 0 ATOM 671 ca ATOM 672 CGI ATOM 673 CG2 ATOM 674 C ATOM 675 0 45 ATOM 676 N ATOM 677 H ATOM 678 CA ATOM 679 CB ATOM 680 CO ATOM 681 ODi ATOM 682 OD2 ATOM 683 C ATOM 684 0 ATOM 685 N ATOM 686 H ATOM 687 CA ATOM 688 CB ATOM 689 CGI ATOM 690 CQ2 6 0 ATOM 691 C ATOM 692 0 ATOM 693 N ATOM 694 H ATOM 695- CA 196 196 196 196 195 196 196 196 196 197 197 197 197 197 198 198 198 198 198 198 198 198 198 198 198 198 198 198 199 199 199 199 199 199 199 199 200 200 200 200 200 200 200 200 201 201 201 201 201 201 201 201 201 202 202 202 202 202 202 202 202 203 203 203 49.928 49.112 $1.193 51.179 51.188 50,972 52.519 51.433 52.442 50.478 49.631 50.704 49.860 50.142 48.830 48.531 48.125 47.870 49.145 49.919 51.123 49.572 50.771 31.567 52.806 53.179 46.794 46.136 46.353 46.951 44.953 44.776 45.617 45.356 44.163 44.489 43.095 42.823 42.321 42.090 41.528 43.399 40.978 40.073 40.787 41.548 39.459 39.501 39.387 38.413 40.251 38.786 39.402 37.561 37.092 36.820 36.274 35.461 37.434 35.669 34.893 35.496 36.163 34.334 74,758 74.264 74.051 72.795 73.150 71,914 73.794 73.661 73.078 73.970 74.363 73.779 72.707 72.320 72.188 72.597 71.016 69.968 69.319 69.969 69.433 68.117 67.572 68,265 67.774 68.383 71.385 72.288 70.714 70.163 70.775 70.400 71.216 72.131 69.810 68.636 70.325 71.240 69.612 70.526 69.691 71.222 69,116 69.889 67.815 67.209 67.259 65,852 65,818 65.246 66.358 67.159 66.593 67.665 68.158 67.390 68.690 68.387 69.634 66.447 66.731 65.309 65.050 64.448 -12.542 -12.313 -12.633 -11.806 -10.326 -9.481 -9.985 -14.123 -14.503 -14.993 -14.695 -16.401 -17.015 -18.140 -16.360 -15.518 -16.872 -15.744 -15.195 -14.251 -13.835 -15.711 -15.298 -14.399 -14.022 -13.376 -17,465 -16.959 -18,533 -19.092 -18.924 -20.402 -21.197 -21.040 -18.077 -17,967 -17.471 -17.708 -16.481 -15.284 -14,142 -14.885 -16,979 -17.248 -17.115 -17,020 -17,292 -17.808 -19,271 -19,737 -19,949 -15.940 -15,030 -15,755 -16.463 -14.543 -13.955 -12.696 -13.649 -14.865 -15.763 -14.183 -13.508 -14.372 27.38 0.00 25.33 24.19 23.30 24.60 31.63 26.23 24.03 27.17 0.0 25.75 26.04 27.75 24.74 0.00 24.17 24.44 17.56 17.53 19.28 16.54 17.99 22.77 26.91 0.00 25.30 28.39 27.93 0.00 29.13 31.52 44.44 0.00 25.88 28.99 22.17 0.00 21.46 17.18 19.27 17.36 25.60 26.95 25.10 0.00 26.46 32.29 40.02 33.36 48.96 28.68 25.09 26.15 0.0 29.60 29.43 26.97 26.15 33.37 36.29 33.12 0.00 28.52 .1

I

I,

i31 'iF 53 ATOM 696 CB ATOM 697 CO ATOM 698 CD ATOM 699 CE ATOM 700 NZ ATOM 701 HZI ATOM 702 HZ2 ATOM 703 HZ3 ATOM 704 C ATOM 705 O0 ATOM 706 N ATOM 707 H ATOM 708 CA ATOM 709 CB ATOM 710 CG ATOM 711 CD ATOM 712 CE ATOM 713 NZ ATOM 714 HZ) ATOM 715 HZ2 ATOM 716 HZ3 ATOM 717 C ATOM 718 O0 ATOM 719 N ATOM 720 R ATOM 721 CA ATOM 722 CE ATOM 723 CG ATOM 724 ODI ATOM 725 ND2 ATOM 726 HD21 ATOM 727 HD22 ATOM 728 C ATOM 729 O0 ATOM 730 N ATOM 731 H ATOM 732 CA ATOM 733 CB ATOM 734 CG ATOM 73s CDI ATOM 736 CD2 ATOM 737 C ATOM 738 O0 ATOM 739 N ATOM 740 H ATOM 741 CA ATOM 742 CB ATOM 743 001 ATOM 744 HOI ATOM 745 C02 ATOM 746 C ATOM 747 O0 ATOM 748 N ATOM 749 H ATOM 750 CA ATOM 751 C ATOM 752 C ATOM 753 O0 ATOM 754 N ATOM 7 5 H ATOM 756 CA ATOM 757 CB ATOM 75 00 ATOM 759 HG

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

ASH

ASH

ASH

ASH

ASH

ASH

ASNH

ASH

ASHN

ASNH

ASNH

LEU

ASN

SN

ASNLE

ASNEU

SN

LEU

LEU

LEU

LEU

LEU

LEU

THR

TIHR

THR

THR

THR

ALA

AA

ALA

A

ALA

ALA

ALA

ALA

SER

SER

SER

SER

58k

SER

203 34.749 63.050 -14.826 203 35.590 62.990 -16.088 203 34,746 63.437 -17.266 203 35.531 63.532 -18.574 203 34.651 64.012 -19.625 203 34.274 64.946 -19.364 203 33.854 63.352 -19.738 203 35.168 64.088 -20.524 203 33.596 64.313 -13.046 203 34.220 64.117 -12.016 204 32.268 64.402 -12.954 204 31.711 64.391 -13.762 204 31.610 64.417 -11.655 204 30.827 65.701 -11.363 204 31.646 66.950 -11.553 204 30.858 68.196 -11.158 204 31.660 69.462 -11.544 204 30.997 70.674 -11.086 204 30.884 70.631 -10.052 204 30.058 70.742 -11.528 204 31.548 71.505 -11.339 204 30.603 63.311 -11.602 204 30.073 62.894 -12.625 205 30.378 62.866 -10.365 205 30.993 63.186 -9.659 205 29.340 61.902 -10.003 205 27.956 62.531 -10.122 205 27.915 63.623 -9.103 205 28.416 63.536 -7.988 205 27.363 64.768 -9.456 205 27.014 64.830 -10.369 205 27.318 65.493 -8.798 205 29.323 60.626 -10.792 203 28.356 60.317 -11.453 206 30.354 59.799 -10.793 206 31.092 59.947 -10.163 206 30.352 58.667 -11.698 206 31.657 58.587 -12.498 206 32.070 59.852 -13.281 206 33.375 59.557 -14.009 206 31.010 60.255 -14.297 206 30.196 57.402 -10.933 206 30.211 57.314 -9.715 207 30.043 56.331 -11.664 201 29.939 56.423 -12.631 207 30.012 55.027 -11.058 207 28.851 54.344 -11.773 207 17.728 54.701 -10.986 207 26.926 54.355 -11.394 207 28.942 52.841 -11.905 207 31.381 54.358 -11.219 207 32.157 54.742 -12.079 208 31.747 53.344 -10.429 208 31.129 53.028 -9.745 208 33.047 52.699 -10.537 208 33.042 51.453 -9.643 208 33.402 52.325 -11.959 208 34.525 52.467 -12.425 209 32.419 51.940 -12.707 209 31.516 51.718 -12.349 209 32,623 51.470 -14.096 209 31.471 50.593 -14.573 209 30.259 51.040 -13.965 209 29.34 50.500 -14.307 28.62 32.11 40.39 50.57 59.39 0.00 0.00 0.00 29.68 30.93 32.55 0.00 34.17 32.07 37.18 44.8 54.50 60.66 0.00 0.00 0.00 32.66 33.79 31.17 0.00 29.81 25.29 26.48 29.97 29.11 0.00 0.00 3034 32.35 31.19 0.00 28.61 29.20 29.46 30.38 33.07 30,76 35.60 32.87 0.00 36.34 36.91 42.24 0.00 42.72 36.64 39.32 36.10 0.00 34.66 32.92 36.07 38.58 38.40 0.00 38.81 45.07 56.06 0.00 i~J, 54 ATOM 760 C ATOM 761 0 ATOM 762 N ATOM 763 R ATOM 764 CA ATOM 765 CB ATOM 766 Co ATOM 767 OD ATOM 768 OD2 ATOM 769 C ATOM 770 0 ATOM 771 N ATOM 772 H ATOM 773 CA ATOM 774 CB ATOM 775 CG ATOM 776 5D ATOM 777 CE ATOM 778 C sc ATOM 779 0 ATOM 710 N SATOM 785 H ATOM 786 CA ATOM 787 CD ATOM 784 001 ATOM 795 HGI ATOM 786 CG2 ATOM 787 C ATOM 788 C ATOM 789 0 ATOM 794 H ATOM 795 CA ATOM 796 CB ATOM 793 OGI 35 ATOM 794 HGI ATOM 795 CG ATOM 300 C ATOM CE0 ATOM 8 CN 40 ATO 7 H ATOM 80 CA ATOM 806 CB ATOM 807 0G ATOM 80 CD ATOM 809 O ATOM 605 CAI ATOM 811 C ATOM 17 CG ATOM 81 N ATOM 809 H ATOM 810 CA ATOM Bi6 CB ATOM 812 CG ATOM 813 CUI ATOM 819 CA2 ATOM 820 C ATOM 816 O ATOM 817 Nl ATOM 8118 H1 ATOM 819 CA ATOM $20 CR ATOM $21- CG ATOM 822 CD ATOM 823 -OI

SER

SEE

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

MET

MET

MET

MET

MET

MET

MET

MET

MET

THR

THR

TIE

HR

ma

THE

THR

THR

THR

THE

THR

THR

THR

TIHR

THR

THR

GLU

ULU

GLU

GLU

ULU

GLIJ

GLU

GLU

GLU

GLU

LEU

LEU

LEU

LEU

LEU

LEU

LFU

LEU

LEU

GLU

GLU

GL

iLU

GLU

GLU

209 209 210 210 210 210 210 210 210 210 210 211 211 211 211 211 211 211 211 211 212 212 212 212 212 212 212 212 212 213 213 213 213 213 213 213 213 213 214 214 214 214 214 214 214 214 214 214 215 215 215 215 215 215 215 215 215 216 216 216 216 116 216 216 32.724 33.571 31.17 31.139 32.061 31.103 29.602 29.209 28.830 33.469 34.155 33.9" 33.371 35.310 35.574 34.914 35.182 34.219 36.310 37.247 36.140 35.445 36.997 36.443 36.580 37.491 37.128 37.122 38.195 35.973 35.126 35.869 34.3S4 33.818 34.273 34.086 36.571 37.304 36.356 35.706 37.082 36.673 35.275 34.998 35.754 34.015 38.561 39.348 38.976 38.327 40.392 40.670 40.608 .82 41.648 40.966 41.998 40.334 39.555 40.802 39.922 39.940 39.089 39.535 52.676 52.733 53.687 53-563 54.930 56.049 55.780 54.945 56.441 55.432 55.935 55.299 54.919 55.693 55.528 56.611 56.360 57.715 54.866 55.382 53.551 53.118 52.704 51.289 50.901 50.999 50.325 53.142 53.275 53.375 53.245 53.796 53.892 52.581 51.966 54.420 55.122 55.298 56.097 55.963 57.351 58.355 58.972 595MS 60.353 59.057 57.147 57.759 56.276 55.881 55.922 54.929 55.-7 54.470 56.656 55.300 55.138 54.261 53.884 53.695 52.537 51.319 50.173 49.546 -14.978 -15.557 -14.779 4.150 -15.516 -15.088 -15.228 -16.050 -14.507 .15.219 -16.096 -13.965 -13.268 -13.629 -12.130 -11.271 -9.493 -8.954 -14.404 -14.999 .14.436 -13.893 -15.242 -15.170 -13.805 -13.313 -16.127 -16.681 -17.241 -17.297 -16.813 -18.681 -19.044 -18.856 -19.436 -20.461 -18.900 -19.865 -18.006 -17.279 -18.103 -17.025 -17.161 -18.559 -19.052 -19.154 -17.960 -18.665 -17.045 -16.422 -16.%42 -15.786 -14.386 .13.329 .14.292 .18.190 -18.686 -13.740 -18.282 -19.991 -20.463 -19-317 .20.064 -21.027 .2 1i 55 ATOM 824 OE2 GLU 216 38.003 49.907 -19.525 1.00 60.44 ATOM 325 C 0LU 216 40.823 54.724 -21.094 1.00 29.88 ATOM 326 O GLU 216 41.722 54.757 -21.926 1.00 27.84 ATOM 827 N ALA 217 39.836 55.617 -21.144 1.00 27.72 ATOM 32 H ALA 217 39.086 5.550 -20.516 1O. o.0o ATOM 829 CA ALA 217 39.351 56.698 -22,121 1.00 27.12 ATOM 830 CB ALA 217 38.530 57.477 -22.045 1.00 27.01 ATOM 831 C ALA 217 41.008 57.620 -21.855 1.00 26.62 ATOM 132 O AIA 217 41.791 57.934 -22.740 .00 26.18 ATOM 833 N PHE 218 41.179 58.083 -2064 1.00 25.15 ATOM 834 H PHE 218 40.554 57.809 -19.917 .00 0.00 ATOM 835 CA PHE 218 42.258 58.999 -20,319 1.00 23.19 ATOM 836 CE PHE 218 42.185 59.322 -13.828 1.00 20.84 ATOM 837 CG PHE 218 43.236 60.340 -18.445 1.00 25.38 ATOM 838 CDI PRE 218 44.361 59.929 -17.742 :.00 24.12 ATOM 339 CD2 PRE 218 43.036 61.686 -18.721 1.00 23.15 SATOM 840 CEI PHE 218 45.234 60.889 -17.2642 1.00 23.97 ATOM 41 CE2 PE 218 43.920 62.634 -18.228 1.00 23.08 SATOM 342 CZ PE 218 45.005 62.234 -17.484 1.00 21.26 20 ATOM 843 C PRE 218 4303 58.427 -20.701 1.00 25.14 ATOM 844 O PHE 213 44.495 59.110 -21.209 1.00 29.00 A,..TOM 345 N ALA 219 43.772 57.138 -20.456 1.00 23.61 ATOM 846 H ALA 219 43.063 56.655 -19.953 o1,00 0.00 SATOM 847 CA ALA 219 44.911 56.429 -20.858 1.00 24.15 25 ATOM 48 CB ALA 219 44.884 54.979 -20.375 1.00 24.62 ATOM 849 C ALA 219 45.224 56.436 22.351 1.00 30.25 SATOM 850 0 ALA 219 46.335 56.291 -22.34 1.00 30.75 ATOM 851 N HIS 220 44.155 56.608 -23.131 1.00 31.80 ATOM 852 11 HIS 220 43.276 56.759 -22.722 1.00 0.00 ATOM 353 CA HIS 220 4.2460 56.691 -24.578 1.00 32.54 ATOM 354 CD HIS 220 43.030 56.108 -25.287 1.00 33.65 ATOM a55 CG HIS 220 43.153 54.627 -25.144 I.00 38.22 ATOM 56 CDZ HIS 220 43.712 53.790 -26.069 3.00 40.87 ATOM 857 NDI HIS 220 42.884 53.94 -24.069 .00oo 2.77 ATOM 858 HDI HIS 220 42.460 54.202 -23.244 1.00 0.00 ATOM 859 CEI HIS 220 43.283 52.673 -24.281 .00 41.06 ATOM 160 NE2 HIS 220 43.788 52.624 -2.483 1.00 39.69 ATOM 861 HE2 HIS 220 44.189 51.815 -25.355 1.00 0.00 SATOM 362 C HIS 220 44.431 58.066 -25.139 1.00 31.27 40 ATOM 63 0 HIS 220 44.482 5.222 -26.346 .00 34.54 ATOM 864 N ARG 221 44.519 59.129 -24.354 1.00 31.23 ATOM 865 H ARG 221 44.453 59.018 -23.381 t 00 0.00 ATOM 866 CA ARG 221 44.684 60.441 -24.935 1.00 25.20 ATOM 867 CB ARO 221 44.496 61.460 .23.34 1.00 23.87 45 ATOM 8s co ARo 221 43.089 63.433 -23.298 .00O 24.28 ATOM 869 CD ARG 221 42.164 62.262 -24.150 1.00 24.77 ATOM 870 NE ARG 221 42.527 63.66 -24.17 100 30.34 ATOM 871 HE ARG 221 43.163 64.003 -24.760 1.00 0.00 ATOM 872 CZ ARG 221 42.025 64.537 -23.214 1.00 36.39 ATOM 873 KHI ARG 221 41.323 64.137 -22.120 1.00 37.62 ATOM 874 HHII ARG 221 41.185 63.163 -21.950 1.00 0.00 ATOM 375 HH12 ARG 221 40.931 64.814 -21.467 1.00 0.00 ATOM 8716 NH2 ARG 221 42.162 65.884 .23.412 1.n 36.36 ATOM 877 HH21 ARG 221 42.631 66.223 -24.228 1.00 0.00 ATOM 878 HH22 ARG 221 41.797 66.529 -22.740 1.00 000 ATOM 879 C ARG 221 46.030 60.643 -25.605 1.00 27.94 ATOM 880 0 ARG 221 47.097 60.475 -25.035 1.00 27.32 ATOM 881 N PRO 222 46.015 61.030 -26.842 1.00 30.14 ATOM 882 CD PRO 222 44.795 61.177 .27.621 I. 28.66 ATOM 883 CA PRO 222 47.20 6.317 -27.635 1.o00 26.80 ATOM 34 CB PRO 222 46.670 61.799 -28.976 300 27.03 ATOM 885 CO PRO 222 45.279 62198 -28.634 L 26.07 ATOM 886 C PRO 222 48.125 62.316 -26.990 100 238.33 ATOM U17 0 PRO 222 49329 62.305 -27.114 1.00 34.07 rI

IB

ii m~ II ATOM 888 N ATOM 889 H2 ATOM 890 CA ATOM 891 CB ATOM 892 CG ATOM 893 CD ATOM 494 OEl ATOM 895 OE2 ATOM 896 C ATOM 897 0 ATOM 898 N ATOM 899 H2 ATOM 900 CA ATOM 901 CB ATOM 902 CG ATOM 903 CD2 SATOM 904 NDI ATOM 905 1DI2 ATOM 906 CEI ATOM 907 NE2 ATOM 908 HE2 ATOM 909 C ATOM 910 O0 ATOM 911 N ATOM 912 H1 ATOM 913 CA ATOM 914 C ATOM 915 CG ATOM 916 CD ATOM 917 CE ATOM 918 NZ ATOM 919 HZI ATOM 920 HZ2 SATOM 921 RZ3 35 ATOM 92 C n ATOM 923 O0 ATOM 924 N ATOM 925 H S 4 ATOM 926 CA ATOM 927 CB ATOM 928 001 ATOM 929 HI01 ATOM 930 CG2 ATOM 931 C 45 ATOM 932 0 ATOM 933 N ATOM 934 H ATOM 935 CA ATOM 936 C6 ATOM 937 o ATOM 938 RG0 ATOM 939 C ATOM 940 O ATOM 941 N ATOM 942 H ATOM 943 CA ATOM 944 CB ATOM 945 CO ATOM 946 D001 ATOM 947 002 ATOM 948 C ATOM 949 0 ATOM 950 N ATOM 951 H 56 47.627 46.669 48.500 47.671 46.658 45.293 44.328 45,.177 49.569 50.606 49.343 48.522 50.346 49.960 48.T74 47.721 48.546 49.236 47.382 46.889 46.004 51.646 52.700 51.608 50.750 52.826 52.503 52.645 52.548 52.571 51.337 50.529 51.211 51.387 53.638 54.854 53.015 52.045 53.803 53.293 51.944 51.452 54.127 53.770 53.809 53.685 53.657 53.733 52.412 52.319 51.584 54.872 55.409 55.246 54.753 56.270 57.305 56.663 57.311 55.524 $5.732 56.509 54.425 53.756 63.254 -24.189 63.259 -26.005 64.265 -21.620 65.306 -24.887 66.030 -25.769 65.381 -25.726 66.114 -25.534 64.168 -25.3880 63.758 -24663 64.371 -24.461 62,599 -24.034 62.108 -24.230 62.037 -23.139 60.670 -22.601 60.779 -21.676 39.901 -21.673 61.704 -20.722 62.337 -20.408 61.423 -20.177 60.352 -20.773 59.939 -20.625 61.845 -23.859 62.029 -23.281 61.474 -25.138 61.290 -25.584 61.359 -25.937 60.985 -27.373 59.492 -27.564 59.057 -29.024 57.524 -29.181 56.917 -28.699 57.292 -29.236 57.140 -27.691 55.887 -28.827 62.627 25.985 62.599 -26.051 63.792 -25.975 63.869 -26.061 64.992 -25.854 66.018 -26.896 65.709 -27.203 66.009 -26A22 65.983 -28.180 65.546 -24.441 66.756 -24.226 64.702 -23.408 63.727 -23.528 65.196 -22.055 64.917 -21.322 65.533 -20.021 65.178 -19.486 64.535 -21.301 63.498 -21.651 65.201 -20.199 66.020 -19.977 64.715 -19.270 65.848 -18.973 67.095 -18.438 68.115 -18.499 61071 .17.976 64.156 -17.923 63.644 -17.121 64.213 -17.599 64.595 -18.2113 25.34 0.00 28.40 27.51 32.04 34.72 35.25 38.35 28.17 28.18 26.29 0.00 25.11 21.67 28.93 29.23 26.59 0.00 31 4 33.43 0.00 26.89 30.62 30.81 0.00 32.77 39.91 55.31 67.66 75.00 78.51 0.00 0.00 0.00 30.17 32.06 29.27 0.00 29.35 28.73 27.05 0.00 31.09 30.98 34.36 27.8S 0.00 25.31 23.69 29.36 0.00 27.92 22.67 28.65 0.00 28.62 21.37 23.40 25.53 24.44 29.79 33.42 30.06 0.00

I

57 ATOM 952 CA ATOM 953 CB ATOM 954 OG ATOM 955 HG ATOM 956 C ATOM 957 0 ATOM 958 N ATOM 959 H ATOM 960 CA ATOM 961 C ATOM 962 OG1 ATOM 963 HGI ATOM 964 C62 ATOM 965 C ATOM 966 0 ec.. ATOM 967 N ATOM 968 H ATOM 969 CA ATOM 970 Ca 20 ATOM 971 C ATOM 972 CDI ATOM 973 CD2 ATOM 974 CEI ATOM 975 CE2 25 ATOM 976 Cz ATOM 971 C ATOM 978 0 ATOM 979 N ATOM 980 H ATOM 982 CA ATOM 982 CB ATOM 983 CI ATOM 984 CDI ATOM 985 CD2 ATOM 986 C *0 4 ATOM 987 0 ATOM 988 N o ATOM 989 H a* ATOM 990 CA ATOM 991 Ca 9 ATOM 992 CGI ATOM 993 CG2 ATOM 994 C ATOM 995 0 45 ATOM 996 N ATOM 997 H ATOM 998 CA ATOMI 999 CS ATOM 1000 CC ATOM 1001 CI ATOM 1002 CD)2 ATOM 1003 CEI ATOM 1001 CE2 ATOM 1005 Cz ATOM t006 c ATOM 1007 0 ATOM 1008 N AM 1009 H ATOM 1010 CA ATOM 1011 C5 ATOM 1012 CG ATOM 1013 SD) ATOM 1014 CE ATOM 1015 C 53.905 63.70 54.314 64.599 54.171 65.969 53472 66.56 52.413 63.747 51.900 64.273 51.720 63.201 52.164 62.618 50.303 63.473 49.501 62.410 48.158 62.903 48.159 63.784 49.485 61.030 49.933 63.471 50.683 63.015 48.751 63.999 48.196 64.345 48.169 64.022 47.955 65.441 49.190 65.978 49.220 66.071 50.273 66.392 50.361 66.561 51.391 66.900 51.442 66.965 46.792 63.409 46061 63.809 46.343 62.479 46.149 62.069 44.915 62.139 4".627 60.672 45.050 60.289 46.436 59.760 44.089 59.40 44.338 62.387 44.9G0 62.002 43.174 62.998 42,629 63.238 42.664 63.90 42.519 64.812 42.185 65.09D 43.826 65.525 41.291 62.645 40.429 62.303 41.049 61.906 41.749 61.801 39.764 61.286 39.87 59.773 40.400 59.110 39.525 5Sa55 41.769 59.045 40.019 57.951 42.262 58.445 41.379 57.910 39.259 61.858 39.990 61.926 38.006 62.298 37.441 62.301 37451 62.780 37.276 64.294 38,619 64.946 38.601 66.742 38,003 66.982 36120 62.112 -16.33 -15.282 -15.,47 -14.792 -16.437 -17.423 -25.435 .14.780 -15.239 -16.052 -16.198 -16.611 -1 .391 -13.739 -12.887 -13.412 -14.142 -12.081 -11.549 -10.873 -9.486 -11.635 -8.861 .11.010 .9.620 -12.160 -13.062 -11.312 -10.654 .11.293 -11.3" -12.995 -12.956 -13.555 -9.941 979 -9.754 -10.535 4.423 43.180 -539 -9.122 -7.176 -6.495 938 6.767 -8.005 49.90 .3.222 -10.046 -9.353 -10.266 -5.650 -4.669 -5.583 4.34 -4326 4.343 4.6u1 4.482 .4.736 2.85 21. 71 21.06 0.00 23.31 23.18 24.93 0.00 22.17 28.90 23.08 0.00 17.19 22.19 29.95 23.0 0.00 22.98 22.01 23.66 22.17 21.19 23.83 18.70 22.93 21.75 22.92 20.85 0.00 19.87 23.90 30.22 32.80 32.58 18.92 22.64 20.69 0,00 22.20 21.39 20.93 27.2 27.10 24.87 24,72 0.00 23.25 20.02 21.08 26.42 2579 29.30 23.33 31.52 25.38 27.17 26.22 O.O0 25.64 24.45 30.66 37.65 33.2 29.25 7P i 58 ATOM 1016 0

MET

ATOM 1017 N 21 ATOM 101 H

SEE

ATOM 1019 CA

SER

s ATOM 1020 Ca

SR

ATOM 1021 OG Sm1 ATOM 122 HS

SER

ATOM 1023 C SEl ATOM 1024 0

SER

ATOM 1025 N

HIS

ATOM 1026 H

HIS

ATOM 1027 CA HiS ATOM 1028 CB Is ATOM 1029 CG

HSL

AToM 1030 Cm HiS ATOM 1031 NOI

HIS

ATOM 1032 HDI

HIS

ATOM 1033 CEI

HIS

ATOM 1034 NEZ

HIS

20 ATOM 1035 H2 HIS ATOM 1036 C IlS ATOM 1037 0

HIS

ATOM 1038 N

GLY

ATOM 1039 H

GLY

AToM o40 CA

GLY

ATOM 1041 C lLY ATM 1042 0

GLY

ATOM 1043 N

ME

ATOM 104 H 11.

ATOM 1O4S C

LE

ATOM 1046 CB 12 ATOM 1047 CG2

ME

ATOM 1048 Cr 1.a 3 ATOM 1049 CDI ME ATOM 1050 C

ILE

ATOM 1051 0

ILE

ATOM 1052 N

ARG

ATOM 101 H

ARG

40 ATOM 1054 CA

AXIS

AMUon Ims ca ARG ATOM os CC ARG ATOM loS? C

ARG

ATOM 1057 CD

ARO

ATOM 1058 NE

ARG

ATOM Ion9 RF

ARG

ATOI 1060 CZ

ARI

ATOM 1061 NHI

ARIG

ATOM 1062 HH11

ARG

ATOM 1063 1112

ARG

ATOM 1064 NHI ARG #,TOM 1065 HMI

ARG

ATOM 1066 HH22

AXIO

ATOM 1067 C

ARG

ATOM 1068 0

AXIS

ATOM1 1069 N GLU ATOM 1070 H

GLU

ATOM 1071 CA

GLI)

ATOM 1072 C

GLU

ATOM 1073 CG 01.

ATOM 1074 CD 5.0.G ATOM 1075 oE0 (1.

ATOM 1076 022

GLU

AMIM 1077 C

OLU

ATOM 10n 0

GLU

ATOM 1079, -1

GLY

2 23 23 23 231 231 23 237 237 237 237 237 237 237 237 237 237 237 237 237 238 238 238 238 239 239 239 239 239 239 239 239 2.39 240 240 240 240 240 240 240 240 240 240 240 240 240 240 240 240 240 240 241 241 241 241 24 1 24, 241 241 241 242 35 35.339 42,136 -3.175 ISMS.80 61.432 -3.138 6 36.441 61 .364 -2.382 6 34.531 60.7,69 -2.995 6 34.412 59.59 -3.971 6 33.M2 58.96 -3.835 6 32.463 59.641 -4.143 34.462 60.246 -1.373 6 35.389 60.340 -0.778 33,329 39.661 -1.203 32,531 59.730 -1,783 33.332 58.80 40015 31.882 58-331 0.377 31.079 59.375 1.141 29.798 59.740 0.816 31,457 60.128 2.188 32.32S 60A120 2.661 30.46 60.934 2.446 29.466 60.721 1.609 28.6S39 61,252 1.540 34.194 57.5"~ .0.321 34.52 57.234 L1462 34.606 SU.82 0.702 34.A29 57.073 1.646 3539 55.668 0.428 M .217 S4,774 1.609 3437 55.199 2.707 3547 53.512 1.370 35.767 S3.23 0.474 3S.461 52.4" 2,439 NA.41 51.45 2.210 33.067 52.124 2.180 34.64S 30.683 0.928 33.745 49.470 0.810 36.822 S1.930 2.435 37.M4 U52. 1.572 37.118 51.076 3.398 36.434 501847 4.063 .38.428 3046 3.465 38.400 49.473 4.647 39.680 41.660 4892 40,939 149.489 5.139 42.114 41.529 4.394 42.319 48.953 3.6W 42.7n2 41.052 5.327 44,059 47M56 4.740 44.2S9 47.111 3.799 44.688 46.986 51247 42.657 4771 6.622 4188 48.0oq 7.073 43.29 47.122 7.115 38.W6 49.791 2.164 40.040 49.8115 1.8"* 37.969 49,138 1.399 37.022 49.233 1.606 38.319 48.416 0.151 37.135 47.44 A0M8 37.559 46.447 -1.379 337 45 .6M6 -2.0$8 35,339 46.31o .2.712 ;6.350 4444) -1.470 W8.S" 49.403 -1.009 39.236 49.10 *-L98 38.015 5062 .966 1.1 1.1 1.0 1.0 3.0 3.0r 1.0( 1.00 1.00 .00 1.00 1.00 1.03 I.00 1.00 .00 .oo .00 .00 1.00

LOD

1.00 1.00 1.00 1.00 1.00 1.00 1.00 L~oo Looa 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 L.oo 1.00 L~oo 1.00 I.0 1.00 1,0 .00 .00 .00 .00 .00 .00 O 31.99 0 0.00 0 32.91 0 31.29 32.86 0.00 32.69 33.35 33.10 0.00 34,12 31.00 40.88 41,19 0.00 43.14 41.94 0.00 33.55 32.97 32.27 0.00 31.94 31.47 29.61 30.61 0.00 36,45 38.77 42.42 42.77 45.44 37.10 37.25 39.29 0.00 42.79 43.84 S2.68 51.36 60.04 0.00 66.92 73.i2 0.00 0.00 65.55 0.00 0.00 43.91 39.72 46.63 49,66 84.57 R6.53 90.34 45.34 453.76 40.7s ~Sggin~re4aaiar~%aa~.~ rcpll~ ~8ass4t~a~e~s~ 3~ 59 ATOM 100 R ATOM 1081 CA ATOM 1082 C ATOM 1083 0 ATOM 1084 N ATOM 1085 R ATOM 1086 CA ATOM 1087 CB ATOM 108U CQ2 ATOM 1089 CGI ATOM 1090 CDI ATOM 1091 C ATOM 109 0 ATOM 093 N ATOM 1094 1 ATOM 1095 CA ATOM 1096 CB ATOM 1097 SG ATOM 18 C ATOM 1099 0 ATOM 1100 N ATOM 1101 H ATOM 1102 CA ATOM 1103 C ATOM i104 0 ATOMlIS N ATOM 1106 H ATOM 1107 CA ATOM 1108 CB ATOM 1109 CG ATOM 1110 CD ATOM I111 CE ATOM 1112 NZ ATUM 1113 HZ1 ATOM 1114 HZ ATOM lI 123 ATOM 1116 C ATOM UP1 0 ATOM 11l N ATOM 1119 H ATOM 1120 CA ATOM 1121 CE ATM 1122 CG ATOM 1123 CD ATOM 1125 ATOM 1124 CE ATOM 1126 HZ1 ATOM 1127 122 ATOM 1128' H3 SO ATOM 1129 C ATOM 1130 0 ATOM 1131 N ATOM 1132 H ATOM 1133 CA ATOM 1134 CD ATOM 1135 CO ATOM 1136 CD.

ATOM 1137 NDI ATOM 1138 HDI ATOM 1139 CEI ATOM 1140 N82 ATOM 1141 HIE2 ATOM 1142 C ATOM 1143 0

GLY

OLY

GLY

GLY

LE

ILE

LE

I.E

ILE

ILE

CYS

CYS

CYS

YS

CYS

CYS

CYS

CYS

GLY

GLY

GLY

GLY

CYS

LYS

LYS

LYS

LYSv

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

HS

HS

HIS

HIS

HS

HIS

HIS

HIS

LS

LYS

LS

LS

1S 242 37.428 50.949 242 38.342 31.556 242 37.375 52.679 242 36.694 52.955 243 37-90 53.311 243 37.107 52.88 243 36.590 54,576 243 37.34 55.376 243 36.739 56.740 243 38.749 55.64 243 39.643 56.0T7 243 35.148 54.304 243 34.920 53.418 244 34.151 55.017 244 34.331 551767 244 32.774 54.760 244 31.955 55.223 244 32.093 54.318 244 32.291 55.415 244 32.421 56.619 245 31.95 54.653 245 31.152 53.680 245 30.973 55.279 245 29.574 55.745 245 29.085 55.433 246 28.868 5601 26 2931 56.669 246 27.604 57.140 246 27.040 57.989 246 26.636 57.098 246 25.882 57.806 246 24.5W 58.422 246 24.007 59.246 246 23.833 58.663 246 24.671 60.006 246 23.112 59.670 246 26.477 56.209 246 25.597 56.538 247 26.479 54.982 247 27.176 4.727 247 25.465 54.045 247 25209 53.130 241 24.508 53.886 247 24143 53.001 247 23M3 53.721 241 23.373 52.835 241 22.808 52.005 247 24.310 52.525 247 22.899 53.347 247 25.856 53251 247 a5.319 52203 248 26.793 53.661 248 27.235 54.526 248 27,155 52.799 248 28.438 53.305 248 28.94 52.330 248 29.411 51.071 248 28.983 52.505 248 28.723 53.315 248 29.467 2 1.412 248 29.720 50.547 248 30-051 49.640 248 2605 52.703 248 23.591 53.685 241 -2.017 -1.071 -3213 .3.992 -3.407 -4.511 -4.856 -3.963 .5.135 -3.806 .618 -3176 -2.666 -3.652 -2.553 -1.030 -4.95 -5.169 -5.563 -5.763 -6.962 .5.470 -7.415 -8.312 -7.033 -3.186 -9.348 -10.459 -10.000 -11.063 -11.905 -11.295 -II'm( -10.742 574 -5.17 -7.019 -7.7 13 -6.685 -7.875 -10220 -11.39 -ILM6 -12.261 -12.261 -13297 -5.450 -5.182 4.603 4C729 .3.498 -2.813 -1.737 -1.973 -0.416 0.00 0.129 -0.817 -0.6m -2.473 -19 0.00 33.55 32.49 31.51 0.00 30.10 28.31 31.33 35.46 30.26 30.91 29.95 0.00 31.93 29.44 36.43 35.60 3.9 34.56 0.00 37.28 42.13 41,53 44.81 0.00 47.47 46.88 51.8 62.88 69.93 76.93 0.00 0.00 0.00 48.40 42.25 45.29 0.00 49.00 54.12 42.74 72.81 81.39 86.55 0.00 0.00 0.00 49.50 51.73 50.46 0.00 45.73 40.60 36.78 33.70 38.30 0,00 34.53 33.3f

O.OD

48.32 44.66 ~a~E~B~W$raa~sasa~

C

Cc Ci

C

C ATOM 1144 N ATOM 1145 II ATOM 1146 CA ATOM 1147 CD ATOM 1148 OG ATOM 1149 HG ATO)M 1150 C ATOM 1151 0 ATOM 1152 N ATOM 1153 1 ATOM 1154 CA ATOM 1155 ca ATOM 1156 CO ATOM 1157 CD ATOM 1151 OEl ATOM 1159 0M ATOM 1160 C ATOM 1161 0 ATOM 1162 N 20 ATOm 1163 11 ATOM 1164 CA ATOM 1165 CB ATOM 1166 CG ATOM 1167 CD 25 AToM 1163 oEI ATOM 1169 N2 ATOM 1170 HMI2 AOMN 1171 11EM2 ATOM 1172 C 30 AToM 1173 0 ATON 1174 N ATOM 1175 H ATOM 1176 CA ATOM 1177 CB ATm 1173 a C ATOM 1179 CG2 ATOM 110 C ATOM 111 0 ATOM 1182 N 40 ATM 113 cD ATOM 1134 CA ATOM IlaS CE ATOM 1186 CG ATOM 117 C ATOM tin o ATOM 1189 N ATOM 1190 11 ATOM 1191 CA ATOM 1192 CB AOMm 1193 CG ATM 1194 ODI ATOM 1195 0D2 ATOM 1196 C ATOM 1197 0 ATOM 1198 N ATOM, 1199 11 ATOM 1200 CA ATOM 1201 CB ATOM 1202 CG2 ATOM 1203 CGI ATOM 1204 CD1 AIOM 125 C ATOM 1206 a ATOM 1207 1

SER

SER

SER

SEl

SER

GLU

GLD

OGL

1LU

OLD

(iLU

GLU

GLU

GLU

OLD

OLD

GLN

OLN

GLN

GLN

OLN

GLN

OLN

~lN OLN

VAL

VA.

VAL

VAL

VA.

VAL

VAL.

VAR

PRO

PRO

PRO

PRO

PRO

PRO

PRO

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ILE

ELE

E.S

ELE

1LE

ILE

lLu VU ILE

LEU

25.626 51.478 25.152 50.711 24.798 S1 23-4U5 50.59 23.488 49.67 23996 49.345 25.626 50.270 26.610 49.341 25.285 50.003 24.519 50.60 25.982 48.958 25.518 48.929 25.746 "0.253 25.05 S.olo 25.196 51.0 5 25.633 48.922 25.7n 47.573 26.649 46.720 24J74 47369 23.971 48.135 24.155 46.078 22.616 4S.902 21.6 47.096 21.754 49.076 22.556 4U05 20.940 47965 20.218 47236 21.031 48.633 24-544 45.330 24.771 44.704 24,66 46.363 24.241 47.718 25.375 46.711 24.482 46,902 25.271 47.177 23.711 45.600 6.466 47.745 26.21 48.961 27448 47.275 28.02 45.V75 28.312 48.113 29905 4751 29.165 46.015 29.113 49.551 28.914 47.837 29.533 49.301 2909 50.376 30-03 5034 29.051 51.461 29.31 51.949 28.990 53.090 29.903 51.204 31.400 50.90 31-50 52.00 32.4g 50.0 32.50 49.50 33.761 50.400 34.181 49317 35.615 49.605 33.157 49,254 32.955 50.50 34.790 50.453 34626 49.556 35.642 51.410 -2.193 -2.770 .04G -1.432 -1.933 -2.757 -0.760 0.982 1.391 1.710 3.169 3.922 5.411 6,085 5.895 1.115 1.119 0.595 0.546 0.085 024 0.015 1.182 1.219 2214 1234 2.26 -1.356 -1.776 -2.165 -1.911 -3.425 4.657 -5.941 -4.932 -3.424 -3,4Q9 -3,427 -3.27 -3.455 -2.798 -22 -5.853 -4.931 -4.143 -6156 4.530 .799 -8.264 -8.796 -5.739 -5.321 -5.937 -C329 .5.307 4.295 -3.797 -3.1" -2.435 428 -7.25 4.503 1.00 1.00 1.00 1.00 1.00 1.00 LOD .00 .00 .00 1.00 .00 .00 .00 .0O 1.00 1.00 1.00 1.00 1.00 .00 .00 .00 1.00 1.00 .00 .oo 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 I bo 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 L~oo 1.00 1.00 IAOo 1.0 1.00 1.00 1.00 1300 .00 I.00 1.00 53.23 0.00 57.14 56.80 58.69 0.00 59.00 55.94 68.05 0.00 77.66 85.81 96.97 103.17 108.23 105.30 77,99 78.08 77.K, 0.00 77.63 6.21 97.44 106.14 112.85 108.36 0.00 0.00 74.59 7325 70.06 0.00 66.24 68.01 67.53 71.77 60.42 57.54 57.04 57.51 53.11 57.01 57.3 47.52 45.35 42.16 0.00 38.48 36.32 36.27 39.87 36.65 35.50 34.98 33.54 0.00 33.65 39.66 40.31 39.28 40.5 39.99 40.26 39.89 vi 61 ATOM 1203 H LEO ATOM 1209 CA LEU ATOM 1210 CE LEU ATOM 1211 CG LEO ATOM 1212 CDI LETO AOM 1213 CD2 LEO ATOM 1214 C LEO ATOM I 215 0 LEU ATOM 1216 N GLN ATOM 1217 H GLN ANTOM 1218 CA GLN ATOM 1219 CB (LN ATOM 1220 CG 17.

ATOM 1221 C! GLN ATOM 1222 OEI G1 ATOM 1223 N4EI GLN o ATOM 1224 HE21 GLN ATOM 12M5 HE22 GLN ATOM 1226 C COLN ATOM 1227 o GLN ATOM 1228 N4 LEJ TOM 129 H LEO ATOM 123 CA LEU ATOM 1231 CB LEO ATOM 123 CG LEU ATOM 1233 CD1 LEU ATOM 1234 Ct2 LEU ATOM 123 C LEO ATOM 1236 0 LEO ATOM i23 N 4 ASN ATOM 1323 H ASN ATOM 1339 CA ASN ATOM 1240 CS ASN ATOM 1241 CO .451 ATOM 1242 001 ASN ATOM 1243 N42 ASN ATOM 1244 HM1 ASN *~ATOM 1245 UD2 ASN ATOM 1246 C ASH ATOM 1247 0 AV ATOM 1248 N ALA ATOM 1249 H ALA ATOM 120 CA AL ATOM 1231 C ALA 45 Amo-= C ALA ATOM 1253 0 ALA ATOM 1254 N ME ATOM 15 H xE ATOM 1256- CA ILE ATOM 1257 CB am ATOM 1255 Coi mi' ATM 1259 CGI ILE ATOM 1260 CDI E ATOM 1261 C am ATOM 1262 0 ME ATOM 1263 N PHE ATM 1264 H PRE ATOM 1365 CA P1E- ATOM 1266 CE PlE ATOM 1267 CO PRE ATOM i:24 CDI PlHE ATOm 1269 CD- PHlE ATOM 1210 CEI PRlE ATOM 4271 cr- PHlE 2S6 35,471 52.284 26 36.831 51,410 256 37.066 32.75 256 33.293 52,842 256 33114 51.825 26 38.377 54145 2 38.022 51.095 256 3.368 51.713 257 38.658 49.977 25 330 49.468 .157 39.395 49.436 257 40 030 4&M02 257 40M720 47.07 257 39.707 46.094 257 33.56 4&361 257 40.013 44.767 257 40697 44-483 257 39195 41.174 27 40.991 5)53 257 41.296 50.526 253 41.747 50.678 258 41.461 50.433 28 42951 51.48 253 43.638 51.714 25 45171 51.726 25 45742 53.133 258 45.495 50,830 258 43.95 50.464 253 44.3 51.56 259 44114 49561 259 43.67 42.856 259 45.197 49.203 259 4&029 48.071 259 45376 46.750 259 44.1&7 46.674 259 4115 4S.710 259 47.0 5 45.857 259 45,632 44.330 259 44.636 4.900 259 45.417 43.535 260 43309 48.952 260 408 49.000 260 42-7n2 48.991 260 41.24 49.011 260 43.243 50.221 260 43.637 50.176 261 43255_ 51.359 261 42.856 51.344 261 43.812 52.619 261 43.53 53.698 261' 44-6W 55-014 263: 42.300 53.941 261 42.175 54.347 2 45.246 S2.443 261 45-594 52.315 262 46.130 51.861 262 45.3 51.647 2621 47.511 51.5%2 263 48.275 50.353 262 43.793 51.855 262 47.919 52.571 262 50.147 52.112 262 48.402 53.50 262 50.619 53.093 .51364 1.00 -7.340 1.00 -3,074 1.00 -8.988 Looa -10.115 1.00 .9 571 1.00 -6.407 1.00 .5.455 1.00 -6715 1.00 -7.488 1.00 -5.992 1.00 .455 1.00 5.487 .OD -5.046 1.00 4126 L.O0 -5.063 i.00 -5.369 1.00 4.52 1.00 6.300 1.00 -7.457 1.00 -5.315 1.00 -4414 1.00 5m L.00 -4.187 1.00 .4015 10 -3.795 1.00 -2.809 1.00 6564" !10 .7.36 1.00 .654 1.00 6.05 110 7.648 1.00 -7.151 ,2'0 -7280--' (ss .7.644 1.00 -7.110 1.( -7.714 1.0 -9.W7 1.00 -9.896 3.00 .9.180 .0 .3.409 1.00 .10312 1.00 -1035% 1.00 .1123 1.00 -12.395 .00 -10.532 3.00 .9.640 1.00 11.027 1.00 .9.884 1.00 10.311 1.00 -9.44 1.00 -8215 1.00 -11.516 1.00 -12.626 1.00 -10.699 1.00 .9.791 I.00 .11.075 1.00 -9.941 IM.0 4.914 1.00 -8.110 3.00 4.831 1.00 .7.279 1.00 -7.974 1.00 0.00 39.66 38.42 37.25 36.68 39.39 3.52 40.24 39.82 0.00 42.81 49.86 62.3 66,77 0,00 0.00 43.83 44.09 45.79 0.00 48.19 62.23 71.72 65.86 80.83 46.26 43.62 41.70 0.00 41.83 46.19 49.87 56.43 50.33 0.00 0.00 38.03 35.47 33.64 0.00 29.46 25.72 28.23 31.28 28.30 0.00 25.63 27.06 2.81 23.10 15.19 27.03 30.14 25.11 0.00 23.29 23.94 22.84 26.517 22.06 29.99 24.85 61 ATOM 1208 IH LEU 256 35.471 52.284 -5.964 1.00 0.00 ATOM 1209 CA LEU 256 36.931 51.410 -7.340 1.00 39.66 ATOM 1210 CB LEU 256 37.06 52.755 -8.074 i.00 38.42 ATOM 1211 CG LEU 256 38.298 52.842 -8.988 1.00 37.25 ATOM 1212 CDI LEU 256 38.214 51.825 -10.115 1,00 36.68 ATOM 1213 CD2 LEU 256 38.377 54.245 -9.571 1.00 39.39 ATOM 1214 C LEU 256 39.022 51.095 -6.407 1.00 38.52 ATOM 1215 0 LEU 256 38.368 51.793 -5.455 1,00 40.24 ATOM 1216 N GLN 257 38.658 49,977 -6.715 1.00 39.82 ATOM 1217 H GLN 257 38.308 49.468 .7.488 1.00 0.00 ATOM 1218 CA GLN 257 39.795 49.436 -5.992 1.00 42.81 ATOM 1219 CB GLN 257 40.050 48.020 -6.455 1.00 49.86 ATOM 1220 CG GLN 257 40.720 47.077 -5.487 1.00 62.03 ATOM 1221 CD GLN 257 39.707 46.054 -5.046 1.00 66,77 ATOM 1222 OEI GLN 257 38.568 46.361 -4.726 1.00 70.03 ATOM 1223 NE2 GLN 257 40.013 44.767 -3.063 1.00 67.84 ATOM 1224 HE21 GLN 257 40.897 44.483 .5.369 1.00 0.00 ATOM 1225 HE22 GLN 257 39.295 44.174 .4.752 1.00 0.00 ATOM 1226 C GLN 257 40.991 50.253 .6.300 1.00 43.83 ATOM 1227 0 GLN 257 41.296 50.526 .7.457 1.00 44.09 ATOM 1228 N LEU 258 41.747 50.678 .5.315 1.00 45.79 ATOM 1229 H LEU 258 41.461 50.433 -4.414 1.00 0.00 ATOM 1230 CA LEU 258 42.951 51.468 .5.556 1.00 48.19 ATOM 1231 CB LEU 258 43.638 51.714 -4.187 1.00 62.23 ATOM 1232 CG LEU 258 45.178 51.726 -4.015 1.00 71,.72 ATOM 1233 CDI LEU 258 45.742 53.135 .3.795 1.00 65.86 ATOM 1234 CD2 LEU 258 45.495 50.830 .2.809 1.00 80.83 ATOM 1235 C LEU 258 43.958 50.864 -6.564 1.00 46.26 ATOM 1236 0 LEU 258 44.583 51.564 -7.368 1.00 43.62 ATOM 1237 N ASN 259 44.214 49.561 .6.654 1.00 41.70 ATOM 1238 H ASN 259 43.687 48.856 .6.205 1.00 0.00 ATOM 1239 CA ASN 259 45.197 49.203 -7.648 1.00 41.88 ATOM 1240 CB ASN 259 46.029 48.071 .7.151 1.00 46.19 ATOM 1241 CG ASN 259 45.376 46.750 -7.280 1.00 49.87 ATOM 1242 ODI ASN 259 44.187 46.674 -7.008 1.00 56.41 ATOM 1243 ND2 ASN 259 46.115 45.710 -7.644 1.00 50.33 ATOM 1244 HD21 ASH 259 47.075 45.857 -7.810 1.00 0.00 ATOM 1245 HD22 ASN 259 45.682 44.830 -7.714 1.00 0.00 ATOM 1246 C ASN 259 44.636 48.900 -9.007 1.00 38.03 ATOM 1247 0 ASN 259 45.417 48.585 -9.896 1.00 35.47 ATOM 1248 N ALA 260 43.309 48.952 -9.180 1.00 33.64 ATOM 1249 H ALA 260 42.708 49.000 -8.409 1.00 0.00 ATOM 1250 CA ALA 260 42.732 48.991 -10.512 1.00 29.46 ATOM 1251 CB ALA 260 41.224 49.011 -10.356 1.00 25.72 ATOM 1252 C ALA 250 43.243 50.221 -11.230 1.00 28.23 ATOM 1253 0 ALA 260 43.637 50.176 .12.385 1.00 31.28 ATOM 1254 N ILE 261 43.255 51.359 .10.532 1.00 28.30 ATOM 1255 H ILE 261 42.856 51.344 -9.640 1.00 0.00 ATOM 1256' CA [LE 261 43.812 52.619 .11.027 1,00 25.68 ATOM 1257 CB [LE 261 43.758 53.698 -9.884 1.00 27.06 ATOM 1258 CG2 ILE 261 44.364 55.014 -10.371 1.00 22.81 ATOM 1259 CGI [LE 261 42.300 53.941 -9.444 1.00 23.10 ATOM 1260 CDI ILE 261 42.175 54.847 -8.215 1.00 15.19 ATOM 1261 C ILE 261 45.246 52.443 -11.516 1.00 27.03 ATOM 1262 0 ILE 261 45.594 52.815 -12.626 1.00 30.14 ATOM 1263 N PHE 262 46.130 51.861 -10.699 1.00 25.11 ATOM 1264 H PHE 262 45.832 51.647 -9.791 1.00 0.00 ATOM 1265 CA PHE 262 47.511 51.582 -11.075 1.00 23.29 ATOM 1266 CB PHE 262 48.275 50.853 -9.941 1.00 23.94 ATOM 1267 CG PHE 262 48.795 51.855 -8,914 1,00 22.84 ATOM 1268 CDI PHE 262 47.919 52.571 -8.110 1.00 26.50 ATOM 1269 CD2 PHE 262 50.147 52.112 -8.831 1.00 22.06 ATOM 1270 CEI PHE 262 48.402 53.570 -7.279 1,00 29.99 ATOM 1271 CE2 PHE 262 50.619 53.093 -7.974 1.00 24.85 ATOM t7 P 24. I 5 ATOM 1272 CZ ATOM 1273 C ATOM 1274 0 ATOM 1275 N ATOM 1276 H ATOM 1277 CA ATOM 1278 CB ATOM 1279 CG ATOM 1280 ODI ATOM 1281 ND2 ATOM 1282 HD21 ATOM 1283 HD22 ATOM 1284 C ATOM 1285 0 ATOM 1286 N ATOM 1287 H ATOM 1288 CA ATOM 1289 CB ATOM 1290 CG ATOM 1291 SD ATOM 1292 CE ATOM 1293 C ATOM 1294 0 ATOM 1295 N ATOM 1296 H ATOM 1297 CA ATOM 1298 CB ATOM 1299 CG ATOM 1300 CDI ATOM 1301 CD2 ATOM 1302 C ATOM 1303 0 ATOM 1304 N ATOM 1305 H ATOM 1306 CA ATOM 1307 CB ATOM 1308 CG ATOM 1309 OD1 ATOM 1310 ND2 ATOM 1311 HD21 ATOM 1312 HD22 ATOM 1313 C ATOM 1314 0 ATOM 1315 N ATOM 1316 H ATOM 1317 CA ATOM 1318 CB ATOM 1319 OGI ATOM 1320 HG1 ATOM 1321 CG2 ATOM 1322 C ATOM 1323 0 ATOM 1324 N ATOM 1325 H ATOM 1326 CA ATOM 1327 CD ATOM 1328 CG ATOM 1329 CD ATOM 1330 CE ATOM 1331 NZ ATOM 1332 14Z1 ATOM 1333 HZ2 ATOM 1334 HZ3 ATOM 1335 C

PHE

PH

PHE

ASN

ASN

ASN

ASN

ASH

ASN

ASN

ASN

ASN

ASH

ASN

MET

MET

MET

MET

MET

MET

MET

MET

MET

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LASNEU

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

ASN

TR

TR

THR

THR

THR

THR

THR

THR

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

262 262 262 263 263 263 263 263 263 263 263 263 263 263 264 264 264 264 264 264 264 264 264 265 265 265 265 265 265 265 265 265 266 266 266 266 266 266 266 266 266 266 266 267 267 267 267 267 267 267 267 267 268 268 268 268 268 268 268 268 268 268 268 268 49.756 53,842 47.573 50.725 48.285 50.994 46,796 49.640 46,219 49.470 46.827 48.683 45.872 47.495 46.483 46.467 47,677 46.416 45,694 45.600 44,728 45.639 46.129 44.947 46.450 49.332 47.043 49.081 45.443 50.203 44.940 50.34S 45.054 50.956 43.742 51.698 42.552 50.779 40.953 51.627 40.447 51.015 46.036 51.979 45.903 52.396 47.044 52,435 47.167 52.116 47.941 53.456 47.844 54.709 46.508 55.432 45.942 :5.833 46.722 56.641 49.365 52.980 50.309 53.724 49.583 51.696 48.832 51.117 50.918 51.154 50.905 49.810 50.208 48.692 50.004 48.725 49.889 47.599 50.148 47.527 49.389 46.885 51.463 51.028 50.834 51.391 52.671 50.496 53.155 50.095 53.325 50.491 54.807 50.073 55.330 51.030 54.960 50.998 55.686 50.139 52.618 49.591 52.606 49.803 51.988 48.543 52.010 48.391 51.228 47.643 50.800 46.427 49.912 45.417 49.406 44.239 48.410 44.674 47.936 43.541 48.737 43.066 47.453 42.877 47,263 43.878 50.013 48.367 -7.209 .12.288 .13.247 -12.270 .11.496 -13.372 -13.119 .12.141 .11.842 -11.517 -11.681 -10.928 -14.673 -15,715 -14.640 .13.807 -15.817 -15.565 -15.835 -15.768 .14.191 .16.362 .17.500 .15.617 .14.695 .16.133 .15.272 .15.452 .14.099 -16.339 .16.157 -16,327 -15.972 .15.737 -16.072 .15.355 .16.103 .17.302 .15.435 .14.492 .15.880 .17.496 -18.480 .17.668 -16.911 -18.965 -18.787 .17.858 .16.964 20.052 -19.933 -21.128 .19.427 -18.464 -20.275 -19.458 .20.194 -19.341 -18.258 .17.483 -17.022 -18.122 -16.766 -20.803 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1,00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1,00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 20.46 27.29 29.20 29.26 0.00 32.07 36.54 48.06 52.97 50.97 0.00 0.00 31.78 30.44 30.71 0.00 31.45 36.38 44.27 51.17 51.48 30.63 25.16 27.60 0.00 23.27 24.95 30.77 35.13 30.86 21.55 21.21 20.49 0.00 21.68 20.66 26.15 30.13 26.18 0.00 0.00 27.52 25.25 29.62 0.00 31.01 31.03 36.10 0.00 28.06 31.70 31.87 34.40 0.00 34.41 38.03 46.78 51.28 59.23 63.39 0.00 0.00 0.00 34.09

A

63 1 ATOM 1336 0 LYS ATOM 1337 N

ASN

ATOM 1338 H ASN ATOM 1339 CA ASN ATOM 1340 CB ASN ATOM 1341 CO

ASN

ATOM 1342 ODI ASN ATOM 1343 ND2 ASN ATOM 1344 HD21 ASN ATOM 1345 HD22 ASN ATOM 1346 C ASN ATOM 1347 0 ASN ATOM 1348 N CYS ATOM 1349 H CYS ATOM 1350 CA CYS ATOM 1351 CB CYS ATOM 1352 SG CYS ATOM 1353 C CYS ATOM 1354 0 CYS ATOM 1355 N PRO ATOM 1356 CD PRO ATOM 1357 CA PRO ATOM 1358 CB PRO ATOM 1359 CO PRO ATOM 1360 C PRO ATOM 1361 0 PRO ATOM 1362 N SER ATOM 1363 H SER ATOM 1364 CA SER ATOM 1365 CB SER ATOM 1366 00G SER ATOM 1367 HO SER ATOM 1368 C SER ATOM 1369 0 SER ATOM 1370 N LEU ATOM 1371 H LEU ATOM 1372 CA LEU ATOM 1373 CB LEU ATOM 1374 CG LEU ATOM 1375 CDI LEU ATOM 1376 CD2 LEU ATOM 1377 C LEU ATOM 1378 0 LEU ATOM 1379 N LYS ATOM 1380 H LYS ATOM 1381 CA LYS ATOM 1382 CB LYS ATOM 1383 CO LYS ATOM 1384 CD LYS ATOM 1385 CE LYS ATOM 1386 NZ LYS ATOM 1387 HZ! LYS ATOM 1388 HZ2 LYS ATOM 1389 HZ3 LYS ATOM 1390 C LYS ATOM 1391 0 LYS ATOM 1392 N ASP ATOM 1393 H ASP ATOM 1394 CA ASP ATOM 1395 CB ASP ATOM 1396 CG ASP ATOM 1397 ODI ASP ATOM 1398 OD2

ASP

ATOM 1399 C

ASP

268 269 269 269 269 269 269 269 269 269 269 269 270 270 270 270 270 270 270 271 271 271 271 271 271 271 272 272 272 272 272 272 272 272 273 273 273 273 273 273 273 273 273 274 274 274 274 274 274 274 274 274 274 274 274 274 275 275 275 275 275 275 275 275 49.735 49.267 49.577 48.012 46.995 46.6S5 46.295 46.896 47.263 46.674 48.130 47.253 49.228 49.948 49.351 49.028 43.971 50.770 51.515 51.237 50.428 52.628 52.694 51.292 53.104 54.259 52.220 51.287 52.617 51.440 50.225 50.053 53.068 53.666 52.826 52.343 53.350 52.250 51.343 50.112 52.168 54.519 54.948 55.108 54.753 56.314- 56.853 58.111 58.619 59.960 61.040 60.925 61.021 61.949 57.348 57.594 57.942 57.514 59.007 60.222 60.849 61.282 60.919 58.618 48,325 -21.979 49.054 -19.948 49.149 .19.020 49.651 -20.366 49.607 -19.263 48.179 -19.067 47.488 -19.988 47.620 -17.892 48.175 .17.168 46.672 -17.821 51.078 -20.767 51.677 .21.370 51.744 .20.458 51.324 -19.931 53.133 -20.g48 54.048 -19.676 55.800 -20.133 53.374 -21.287 54.159 -20.711 52.761 -22.313 52.006 -23.263 52.860 -22.730 51.997 -23.974 52.036 -24.514 54.281 -22.955 54.644 -22.776 55.180 -23.365 54.942 -23.574 56.555 -23.545 57.305 -24.135 56.635 -23.790 56.659 -22.840 57.208 -22.269 58.259 -22.345 56,687 .21.064 55.839 -20.921 57.384 -19.910 57.567 -18.851 58.788 -19.139 58.798 -18.225 60.056 -18.965 56.632 -19.336 56.803 -18,205 55.731 -20.112 55.544 -21.008 55.053 -19,684 54.212 -20.819 53.417 -20.485 52.783 -21.777 52.073 -21.658 53.035 -21.669 53.685 -20.867 53.571 -22.560 52.537 -21.5S0 56.087 -19.285 57.057 -19.983 55.855 -18.123 55.152 -17.599 56.689 -17.546 56.827 -18.507 55.482 -18.831 55.358 .19.970 54.577 -17.980 58.090 .17.114 1.00 1.00 1.00 1.00 1,00 1.00 L,00 1,00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 34.97 29.59 0.00 24.54 26.50 32.19 35.32 35.76 0.00 0.00 27.02 27.85 26,07 0.00 23.70 22.29 27.75 26.34 24.34 26.04 26.80 26.32 25.30.

24.28 29.26 31.29 30.94 0.00 31.44 37.05 53.17 0.00 30.10 30.70 29.16 0.00 26.90 23.25 24.56 20.80 20.75 28.09 24.97 28.29 0.00 27.01 28.08 32.44 40.58 45.30 47.25 0.00 0.00 0,00 27.81 27.52 26.78 0.00 24.40 24.70 28.62 35.31 31.34 21,07

'I

Li .1~ 64 ATOM 1400 0 ATOM 1401 N ATOM 1402 H ATOM 1403 CA ATOM 1404 CB ATOM 1405 CO ATOM 1406

CD

ATOM 1407 CE ATOM 1408 NZ ATOM 1409 HZI ATOM 1410 HZ2 ATOM 1411 HZ3 ATOM 1412 C ATOM 1413 0 ATOM 1414 N ATOM 1415 CD ATOM 1416 CA ATOM 1417 CB ATOM 1418 CC 20 ATOM 1419 C ATOM 1420 0 ATOM 1421 N ATOM 1422 H ATOM 1423 CA ATOM 1424 CB ATOM 1425 CG ATOM 1426 CD ATOM 1427 CE ATOM 1428 NZ ATOM 1429 HIZl ATOM 1430 HZ2 ATOM 1431 HZ3 ATOM 1432 C ATOM 1433 0 35 ATOM 1434 N ATOM 1435 H ATOM 1436 CA SATOM 1437 CB ATOM 1438 CGI 40 ATOM 1439 CG2 ATOM 1440 C ATOM 1441 0 ATOM 1442 N ATOM 1443 H ATOM 1444 CA ATOM 1445 CB ATOM 1446 CG2 ATOM 1447 CGI ATOM 1448 CDI ATOM 1449 C ATOM 1450 0 ATOM 1451 N ATOM 1452 H ATOM 1453 CA ATOM 1454 CB ATOM 1455 CG2 ATOM 1456 CGI ATOM 1457

CDI

ATOM 1458 C ATOM 1459 0 ATOM 1460 N ATOM 1461

H

ATOM 1462 CA ATOM 1463. CB

ASP

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

PRO

PRO

PRO

PRO

PRO

PRO

PRO

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

LYS

VAL

VAL

VAL

VAL

VAL

VAL

VAL

VAL

ILE

ILE

ILE

II.E

ILE

ILE ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

ILE

Ile 275 59.450 58.931 -16.787 276 57.332 58.409 -17.087 276 56.635 57.754 .17.318 276 56.918 59.738 .16.690 276 55.977 60.268 .17.772 276 56,700 60,352 -19.129 276 55.762 60.741 -20.275 276 56.341 60.584 .21.682 276 55.317 60.858 .22.677 276 54.962 61.828 -22.562 276 54.528 60.192 -22.550 276 55.716 60.744 -23.630 276 56.238 59.664 -15.339 276 55.549 58.696 -15.080 277 56.342 60.565 -14.432 277 57.125 61.779 -14.555 277 55.658 60.491 -13.158 277 56.178 61.665 -12.348 277 57.365 62.160 .13.113 277 54.146 60.506 .13.252 277 53.522 61.268 .13.988 278 53.531 59,642 .12.473 278 54.049 58.986 .11.955 278 52.093 59.598 -12.436 278 51.654 58.233 -12.953 278 52.074 58.090 .14.435 278 51.919 56.673 -14.945 278 53.057 55.723 .14.550 278 54.258 55.975 -15.318 278 54.573 56.952 -15.158 278 54.052 55.841 -16.328 278 55.008 55.317 -15.025 278 51.698 59.807 -11.016 278 51.903 38.916 .10.202 2719 51.133 60.956 -10.655 279 50.893 61.619 .11.338 279 50.830 61.249 -9.270 279 51.262 62.701 -8.960 279 51.042 63.041 -7.484 279 52.737 62.882 -9.318 279 49.340 61.054 -9.066 279 48.520 61.647 .9.770 280 48.946 60.212 -8.099 280 49.628 59.772 -7.543 280 47,535 59.939 -7.830 280 47.305 58.401 -7.914 280 45.900 58.000 -7.454 280 47.519 57.971 .9.377 280 48.045 56.562 -9.513 280 47.150 60.496 -6.467 280 47.789 60.204 -5.464 281 46.108 61.318 -6.363 281 45.542 61.495 -7.148 281 45.770 61.940 -5.109 281 45.858 63.465 -5,300 281 45.338 64.229 -4.081 281 47.333 63.810 -5.516 281 47.627 65.269 -5.781 281 44.389 61.476 -4.737 281 43.495 61.498 .5.568 282 44.128 61.032 -3.509 282 44.833 61.012 -2.823 282 42.781 60.593 -3.163 282 42.761 59.050 -3.045 1.00 1.00 1,00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 2.00 1.00 1.00 1.00 1.00 1.00 1.00 1,00 1,00 1.00 1,00 1.00 1.00 1.00 1,00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1,00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 22.30 20.11 0.00 20103 15.62 20.79 25.10 24.43 26.93 0.00 0.00 0.00 21.33 23.74 18.25 20.11 19.61 19.47 17.30 21.46 24.07 19.22 0.00 18.47 16.94 15.20 16.03 18.36 19.84 0.00 0.00 0.00 21.76 23.16 18.92 0.00 16.68 15.16 12.89 13.20 20.13 20.96 20.07 0.00 20.53 25.21 17.77 24.13 30.13 21.62 22.51 20.14 0.00 19.43 19.82 16.34 24.03 26.46 22.29 21.61 23.39 0.00 24.32 21.45 wI 65 ATOM 1464 CG2 1E ATOM 1465 CI ILE ATOM 1466 CD IE ATOM 1467 C 1.

ATOM 1469 0 IL ATOM 1469 N GLN ATOM 1470 H GLN ATOM 1471 CA GLN ATOM 1472 CB GLI ATOM 1473 C GLN ATOM 1474 CD GLN ATOM 1475 OE GLN ATOM 1476 NE2 GLN ATOM 1477 HE21 0LN ATOM 1478 HE22 0LN ATOM :479 C GLN ATOM 1480 0 GLN ATOM 1431 N ALA ATOM 1482. H ALA ATOM 1483 CA ALA ATOM 1484 CB ALA ATOM 1485 C ALA ATOM 1486 O ALA ATOM 1487 N ALD) ATOM 1488 CA ALd ATOM 14189 C ALD ATOM 1490 0 AL) ATOM 1491 CB ALD ATOM 1492 S0 ALD ATOM 1493 HI ALD ATOM 1494 CI ALD ATOM 1495 CS ALD ATOM 1496 02 ALD ATOM 1497 C9 ALD S 35 ATOM 1498 CG ALD ATOM 1499 CDI ALD ATOM 1500 CD2 ALD ATOM 1501 CEI ALD ATOM 1502 CE2 ALD) ATOM 1503 CZ ALD ATOM 1504 OH AL!) ATOM 1505 N2 ALD ATOM 1506 C2 AL ATOM 1507 C6 ALD ATOM Isos 03 AL ATOM 1509 C1 ALD ATOM 1510 CGI ALD ATOM 1512 CG2 ALD ATOM 151 N3 ALD ATOM 1513 C3 ALD ATOM 1514 C7 ALD ATOM 1515 04 ALD ATOM 1516 C11 ALD ATOM 1517 N4 AL! ATOM 1518 C4 AL! ATOM 1519 Ca ALD ATOM 1520 C12 ALD ATOM 1521 C13 ALD ATOM 1522 ODI1 ALD ATOM 1523 OD!2 AL) ATOM 1524 05 AD ATOM 1525 HI ALD SATOM 1526 CI4 A d ATOM 1527 HS AL) 282 282 282 232 282 283 283 283 283 283 283 263 283 283 283 283 283 2384 284 284 284 284 284 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 285 41.368 58.517 43.224 58.492 43.645 57.044 42.275 61.214 42.837 61.022 41.195 61.985 40.880 62.229 40.417 62.361 39.916 63.809 38.790 64.295 39.213 64.338 40.263 64.820 38.357 63.923 37.468 63.628 38.632 63.911 39.227 61.397 38.396 61.282 39.127 60.692 39.827 60.742 38.115 59.682 38.196 58.595 38.438 59.050 39.598 58.871 37.396 58.754 37.538 58.081 38.038 56.689 37.779 56.155 36.215 57.908 35.603 59.475 323516 69.905 33.306 68.911 32.529 67.649 31.833 67.247 34.623 68.691 35.549 69.731 35.586 71.002 36.268 69.456 36.376 7L.984 37.056 70.446 37.109 71.725 37.756 72.792 32.657 67.034 31.932 65.809 33.033 64.915 34.011 65.383 30.819 66.116 30.399 64.915 29.588 65.547 32.851 63.630 33.824 62.646 33.316 62.147 32.114 62.219 33.899 61.464 34.265 61.650 34.210 61.553 34.763 60.201 35.030 62.710 34.835 62.936 35.766 63.453 33.772 62.594 33.751 59,337 36.497 58.977 31.983 70.803 31.384 69.885 -2.706 4.387 4.387 .1.878 -0.802 -1.981 -2.879 0.807 .0.951 .0.003 1.454 1.835 2.370 2.058 3.299 -0,.673 -1.530 0.460 1.148 0.688 -0.3a3 2.029 2.387 2.795 4.104 3.860 2.190 4.846 5.487 10.351 10.160 10.069 10.983 10.902 10.350 10.923 9.185 10.326 8.595 9.163 8.546 8.925 8,102 8.206 7.657 7.661 6.822 8.453 8.422 7.997 6.681 6.431 8.952 5.98 4.464 4.008 3.971 2.5310 1.898 1.993 3.478 2.418 10.042 11.844 1.00 1.00 1.00 1.00 I.Co 1.00 1.00 1.00 1.00 1.00 LooO 1.00 1.00 1.00 1.00 1.00 1.00 2.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 .Oo 1.00 1.00 1.00o 1.00 1.00 1.00 1.00 Itoo 1.00 1.00

LODO

1.00 1.00 2.00 1.00 .00o 1.00 1.00 .00oo 1.00 2.00 I.o00 1.00 .00oo 1.00 2.00 1.00 1.00 .00 2.00 I.00 1.00 1.00 2.00 1.00 1.00 1.00 2.00 2.00 22.39 17.29 18.62 24.17 23.02 24.17 0.00 26.94 29.78 30.76 31.96 26.84 32.78 0.00 0.00 27.75 27.45 24.57 0.00 22.92 15.36 25.81 29.97 27.43 26.59 26.61 30.02 -26.43 34.69 58.66 48.23 45.89 47.20 46.18 50.40 53.28 53.34 55.15 52.67 57.31 63.58 38.70 34.11 33.25 27.84 39,67 44.49 40.46 31.01 33.50 37.64 44.05 31.45 35.48 33.98 36.52 3504 37.05 37.30 34.87 42.82 0.00 65.26 0.00 p' j jr a--

I_

r 6 ATOM 1528 H6 ALD ATOM 1529 H7 MD ATOM 1530 H8 ALD ATOM 1531 H9 ALD ATOM 1532 CIs ALD ATOM 1533 01 ALD) ATOM 1534 H2 AL) ATOM 1535 N ARG ATOM 1536 H ARG ATOM 1537 CA ARG ATOM 1538 CB ARG ATOM 1539 CG ARG ATOM 1540 CD ARG ATOM 1541 NE ARG ATOM 1542 HE ARG ATOM 1543 CZ ARG ATOM 1544 NHI ARG ATOM 1545 HH11 ARG *ATOM 1546 HH12 ARG ATOM 1547 NH12 ARG ATOM 1548 HH21 ARG ATOM 1549 1H22 ARO ATOM 1550 C ARG ATOM 1551 O ARG ATOM 1552 N GLY ATOM 1553 H GLY ATOM 1554 CA GLY ATOM 1555 C GLY ATOM 1556 -0O GLY ATOM 1557 N ASP ATOM 1558 H ASP ATOM 1559 CA ASP ATOM 1360 CB ASP ATOM 1561 CG ASP ATOM 1562 ODI ASP ATOM 1563 OD2 ASP ATOM 1564 C ASP ATOM 1565 0 ASP ATOM 1566 N SER ATOM 1567 H SER ATOM 1568 CA SER ATOM 1569 CB SER ATOM 1570 G00 SER ATM 1571 HG SER11 ATOM 1572 C SER ATOM 1573 0, SER ATOM 1574 N PRO ATOM 1575 CD PRO ATOM 1576 CA PRO ATOM 1577 CB PRO ATOM 1578 CG PRO ATOM 1579 C PRO ATOM 1580 O PRO ATOM 1581 N GLY ATOM 1582 H GLY ATOM 1583 CA GLY ATOM 1584 C GLY ATOM 1585 O OLY ATOM 1586 N VAL ATOM 1517 H VAL ATOM 1588 CA VAL ATOM 1589 CB VAL ATOM 1590 CGI VAL ATOM 1$91 CG2 VAL 285 37.739 72.711 285 33.249 67.364 285 32.004 63.310 285 35.098 61.322 285 30.476 70.799 285 32.684 71.569 285 35.481 60.411 286 38.740 56.061 286 38.960 56.493 286 39.231 54.719 286 40.747 54.692 286 41.387 55.701 286 42.430 54.961 286 43.768 55.251 286 43,.926 55.944 286 44.798 54.588 286 46.050 55.018 286 46.181 55.814 286 46.845 54.538 286 44.621 53.491 286 43.700 53.16i 286 45.416 53.010 286 38.629 53.755 286 39.160 52.683 287 37.495 54.113 287 37.021 54.925 287 36.905 53.350 287 36.162 54.330 287 35.932 55.475 288 35.804 53.848 288 36.263 53.019 288 34.941 54.525 288 33.839 53.609 288 33.043 52.985 288 33.051 51.752 288 32.413 53.731 288 35.710 55.003 288 35.283 55.884 289 36.884 54.437 289 37.321 53.757 289 37.596 54.805 289 38.704 53.801 289 38.083 52.535 289 38.773 51.861 229 38.069 56.239 289 38.394 56.809 290 38.143 56.885 290 37.408 56.497 290 38.631 58.270 290 38.051 58.769 290 37.920 57.494 290 40.138 58.521 290 40.637 59.638 291 40.904 57.44 291 40.473 56.576 291 42.342 57.513 291 43.153 57.845 291 44.292 58.236 292 42.646 57.715 292 41,794 57.243 292 43.348 58.171 292 42.800 59.510 192 43.027 60.64 292 41.311 59.404 7.587 8.179 8.842 6.347 10.002 9.386 3.215 4.793 5.651 4.550 4.682 3.739 2.926 3.356 4.031 2.845 3.144 3.735 2.774 2.058 1.851 1.689 5.538 5.803 6.133 5.843 7.208 8.100 7.728 9.292 9.545 10.273 10.820 9.706 9.612 8.944 11.500 12.250 11.775 11.200 12.969 13.071 13.042 13.003 12.940 11.907 14.041 15.249 14.136 15.454 16.266 14.026 13.939 14.034 14,160 13.835 15.056 14.877 16.287 16.414 17.491 18.102 17.151 18.371 0.00 0.00 0.00 0.00 65.03 73.24 0.00 26.38 0.00 25.85 24.02 23.75 27.07 29.64 0.00 26.29 25.89 0.00 0.00 19.53 0.00 0.00 26.43 27.12 26.73 0.00 27.46 30.03 32,06 33,55 0.00 32.84 40.57 48.86 53.99 51.26 31.98 32.42 28.78 0,00 24.44 22.45 33.26 0.00 24.99 26.10 27.20 22.52 25.66 27.04 21.06 28.40 26.84 24.70 0.00 27.48 26.97 24.56 24.36 0.00 26.77 24.74 28.54 25.92 4- 67 ATOM 1592 C ATOM 1593 0 ATOM 1594 N ATOM 1595 H ATOM 1596 CA ATOM 1597 CB ATOM 1598 CGI ATOM 1599 CG2 ATOM 1600 C ATOM 1601 0 ATOM 1602 N ATOM 1603 H ATOM 1604 CA ATOM 1605 CB ATOM 160W6 CG SATOM 1607 CD2 ATOM 1608 CE2 ATOM 109 CE3 S ATOM 1610 CDI S 20 ATOM 1611 NEI ATOM 1612 HEI ATOM 1613 CZ2 ATOM 1614 CZ3 SATOM 1615 CH2 S 25 ATOM 1416 C S* ATOM 1617 0 ATOM 1618 N ATOM 1619 H ATOM 1620 CA ATOM 1621 cI ATOM 1622 CO SATOM 1623 CDI ATOM 1624 CD2 ATOM 1625 CEI 35 ATOM 1626 CE2 ATOM 1627 CZ ATOM 1628 C SATOM 1629 O0 ATOM 1630 N .n 40 ATOM 1631 H ATOM 1632 CA ATOM 1633 CB ATOM 1634 CG ATOM 1635 CD ATOM 1636 CE ATOM 1637 NZ ATOM 1638 HE ATOM 1639 HZ22 ATOM 1640 HZ3 ATOM 1641 C ATOM 1642 O0 ATOM 1643 N ATOM 1644 H ATOM 1645 CA ATOM 1646 CB ATOM 1647 CO ATOM 16498 OD ATOM 1649 OD2 ATOM 1650 C ATOM 1651 0 ATOM 1652 OT ATOM 1653 CB ATOM 1654 C ATOM 1655 O0 292 43.273 57.168 292 42.441 56.276 293 44.199 57.326 293 44.833 58.079 293 44.287 5.466 193 45.426 55.456 293 44.801 55.914 293 45.069 54.152 293 44.566 57.485 293 45.149 58.538 294 44.153 57.245 294 43.657 56,424 294 44.490 58.156 294 43.553 58.075 294 42.160 58.474 294 41.796 59,897 294 40.345 59.680 294 42.296 61.171 294 41.128 57.603 294 40.077 58.356 294 39.189 57.990 294 39.526 60.761 294 41.421 62.218 294 40.084 62.017 294 45.823 57.790 294 46.217 56.632 295 46.574 58.751 295 46.375 59.702 295 47.704 58.370 295 49.007 58.377 295 49.439 59.770 295 50.325 60.433 295 48.853 60.419 295 50.559 61.777 295 49.086 61.764 295 49.928 62.441 295 47.765 59.359 295 47.343 60.503 296 43.314 58.905 296 48.708 58.012 296 48.421 59.737 296 48.497 58.862 296 48.184 59.668 296 47.966 58.760 296 49.201 57.947 2%96 50.320 58.836 296 50.088 59.472 296 50.511 59.398 296 51.159 58.270 296 49.684 60.547 296 50.729 59.951 297 49.591 61.860 297 49.170 62.489 297 50.160 62.460 297 51.698 62.430 297 52.196 61.867 297 51.766 62.345 297 53.019 60.945 297 49.705 63.893 297 49.463 64.419 297 49.571 64.477 317 65.517 45.642 317 63.053 45.718 317 62.644 46.858 18.641 18.749 19.569 19.525 20.719 20.369 20.801 21.025 21.845 21.595 23.098 23.290 24.193 25.389 25.030 24.914 24.556 25.019 24.792 24.518 24.324 24.347 24.795 24.478 24.766 24.801 25.251 25.116 26.063 25.207 24.852 25.679 23.779 25.480 23.583 24.444 27.208 27.145 28.316 28.341 29.496 30.739 31.983 33.189 33.534 33.780 34.569 32.925 34.015 29.397 29.161 29.569 28.934 30.750 30.748 32.082 33.140 32.067 30.706 29.608 31.777 31.211 -30.743 -30.593 26.43 28.53 27.36 0.00 28.84 27.36 23.55 28.31 30.19 30.10 27.38 0.00 26.27 24.27 26.12 27.34 25.56 28.09 25.19 28.67 0.00 21.74 27.80 25.43 27.42 26.36 32.18 0.00 41.42 40.27 44.54 45.58 48.03 49.71 47.03 52.74 44.22 42.87 57.30 0.00 64.18 65.20 68.41 72.60 75.23 81.49 0.00 0.00 0.00 68.26 72.09 72.66 0.00 82.26 88.06 92.95 94.73 96.27 84.91 87.39 84.32 60.98 35.95 55.33 r i i

'I

i

I

r a d g 68 ATOM 1656 HTI ATOM 1657 HT2 ATOM 165 N ATOM 1659 3 SATOM IG6 CA ATOM 1661 N ATOM 2662 H ATOM 1663 CA ATOM 1664 Ca ATOM 1665 CG2 ATOM 1666 CGI ATOM 1667 CDI ATOM 1668 C ATOM 1669 0 SATOM 1670 N ATOM 1671 H ATOM 1672 CA ATOM 1673 Ca ATOM 1674 CG ATOM 3675 CD ATOM 1676 CE ATOM 1677 NZ ATOM 1678 HZI ATOM 1679 Ca 25 ATOM i680 Hz3 ATOM 1681 C ATOM U162 0 ATOM 1683 N ATOM 1634 H ATOM 1685 CA ATOM 166 CS ATOM 1687 CG ATOM 1688 CD ATOM, 1689 CE ATOM 1690 HZ ATOM 1692 HZI ATOM 1692 HZ2 ATOM :1693 HZ3 ATOM 2694 C 40 ATOM 1695 o ATOM 1696 N ATOM 1697 H ATOM 1698 CA ATOM 1699 Ca ATOM 1700 c ATOM 1701 0 ATOM 1702 N ATOM 1703 H ATOM 1704 CA SO ATOM 1705 Ca ATOM 1706 CG ATOM 2707 CD2 ATOM 1708 NDI AMM 1709 HV! ATOM 1720 CEI ATOM 711 NE2 ATOM 271 E22 ATOM 1713 C ATOM 2714 0 TO 'AMTOM 1715 N ATOM 17t6 H AITOM 17 CA ATOM 1718< Ci -ATOM 1719 CG2

ALA

ALA

ALA

ALA

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11.Y

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LYS

LYS

LYS

LYS

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LY

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LYE

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LYE

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HIS

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ILE

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'IL

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11.

317 63.898 317 62.379 317 6.865 317 64.4 317 64.124 318 62.584 328 62.8M 318 61.755 318 60.789 318 59.718 318 61.523 318 60.596 328 62.702 328 63.754 319 62.351 319 61.527 319 63.202 319 63.641 329 65.100 319 65.488 329 66.995 319 67.163 319 66.878 319 66.80 329 68.294 319 62.157 319 60.90 320 62.635 320 63.71 320 61.123 320 62.389 320 62.391 320 63.120- 320 63.108 320 64.002 320 63.689 320 64.966 320 63.995 320 61.701 320 62.635 321 60.521 321 59.T77 321 60.307 322 59.387 321 59.653 321 58.914 322 59.889 322 60.599 322 59.185 322 59.573 322 61.001 322 61.504 322 62.036 322 62.980 322 63.132 322 62.800 322 63.431 322 57.741 322 57,445 323 56.837 323 57,138 323 55.418 323 S4 711I 323 54.71 323 34.53 47.390 46.419 46.499 46.464 45.449 44.702 43.777 44.926 43.749 43.915 42.407 41,194 45.020 44.390 45.808 46.341 45.893 47.280 47.492 48.878 49.115 50.131 49.820 51.037 50.246 45.597 45.806 45.096 44.830 44.993 43.928 42.526 41.594 40.150 39.368 39.403 39.751 38.382 46.280 47.066 46.503 45.8970 47.641 48.636 47.109 46.243 47.690 48.347 47.349 48.206 43.032 46.919 48.845 49.784 43.250 47.082 46.421 47.676 48.725 46.786 45.908 47.059 45.714 45.378 32.203 -3.0d2 -32,753 -33.540 -31.79 -30.23S -28,863 -28.729 -29796 -28.849 -28.946 -27.656 -27.651 -26.630 -26,691 -25,383 -23.528 -24.982 -24.57 -23,739 -2282 -24.022 -23.64 -24.329 -24.324 -23.105 -23.064 _21.m9 -20.978 -21.349 -20.583 -21.065 -20.235 -19.245 -20.319 -2o.57 -21.053 -20.917 -20.61 20.580 -19.592 -20.248 -18.320 -18,310 -17.150 -17,216 15.904 -14.672 -14.197 -13.586 .1c.409 -14.69 -14001 .23.515 -13.150 ,16.169 -16.715 -IS.79 -15.473 435.946 25.710 -14.233 1.00 1.00 1.00 .00 1.00 100 1.00 2.00 2.00 1.00 1.00 2.00 1.00 L~oo 1.00 1.00 1.00 1.00 .00oo 1.00 1.00 2.00 I.00 2.00 1.00 1.00 1.00 1.00 1.00 1.00 I.oci 1.00 1.00 2.00 3.00 1.00 1.00 1.0 3.00 1.00 1.00 1.00 1.00 2.00 2.00 1.00

.OD

3,00 3.00 1.00 1.00 3.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 3.00 3.00 O.00 0.00 63.55 0.00 59.35 52.67 0.00 48.35 43.28 39.84 35.21 36.89 48.70 46.83 47.06 0.00 42.30 46.07 59.38 73.60 83.83 85.82 0.00 0.00 0.00 36.54 36.02 34.11 0.00 35.41 361.25 40.56 40.42 43.28 52.04 0.00 0.00 0.00 33.67 31.18 28.86 0.00 26.84 26.05 25.85 29.15 25.43 0.00 20.62 28.27 16.95 23.40 20.74 0.00 17.22 29.50 000 23.43 21.27 27.07 0.00 25.91 23.73 19.80 .1

-SW

-69 ATOM 1720 CGI ATOM 1721 CDI ATOM IM722 C ATOM 1723 O0 ATOM 1724 N ATOM 1725 H ATOM !726 CA ATOM 1727 C ATOM 1721 CG ATOM 1729 CD ATOM 1730 OEl ATOM 1731 OE2 ATOM 31732 C ATOM 1733 O0 ATOM 1734 N S* ATOM 1735 H ATOM 1736 CA SATOM 1737 CD ATOM 173 CG ATOM n1739 CD ATOM 1740 CE ATOM 1741 NZ ATOM 1742 HZI ATOM 1743 122 25 ATOM 1744 HZ23 ATOM 1745 C ATOM 1746 O0 ATOM 1747 N ATOM 1748 H1 ATOM 1749 CA ATOM 1750 CB ATOM 1751 CGo ATOM 1752 _ODI ATOM 1753 OD2 ATOM 154 C ATOM 1755 0 ATOM 1756 N ATOM 17S7 H ATOM 1758 CA 40 ATOM 1759 CD ATOM 1760 CG ATOM 1761 CDI ATOM 1762 CD2 ATOM 1763 CEI ATOM 1764 CE2 ATOM 1765 C ATOM 1766 C ATOM 1767 O0 ATOM 1768 N ATOM 1769 H ATOM 1770 CA ATOM 1771 C ATOM 1772 CG2 ATOM m177 CI ATOM 1774 CDI ATOM 1775 C ATOM 1776 O0 ATOM 1777 N ATOM 778 H ATOM 2779 CA ATOM 15 CBa ATOM 1781 C ATOM 1782 0 ATOM 1M3 N

IE

ILE

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MLE

GLU

GLU

GLU

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ALA

PHE

323 53.41l 45.795 323 52.681 44.478 323 54.891 48.183 323 S3.998 48.930 324 55.418 48.357 324 56.140 41.768 324 54.973 49,441 324 54.186 48.824 324 53.692 49.837 324 52.881 49.122 324 52.25 48.099 324 52.858 49.586 324 56.191 50.205 324 57.106 49.587 325 56.245 51,531 325 55.526 52.005 325 57.371 52.335 325 58.571 52.036 325 59.855 52.715 325 61.087 52.259 325 61.101 52.703 325 61.146 54.143 325 62.010 54.507 325 60.311 54.557 325 61.145 54.397 325 56.161 53.774 325 55.928 54.033 326 57.480 54.700 326 58.191 54AIS.415 326 57.214 56.136 326 57.422 56,737 326 58.721 56.273 326 59.506 56.575 326 58.646 55.579 326 55.835 56.529 326 55.284 57.559 327 55.222 55.721 327 55.626 54.862 327 53.972 56.107 327 53.008 54.938 327 52.290 54.444 327 51.070 55.004 327 52.772 53.338 327 50.334 54.424 327 52.022 52.752 327 50.801 53.298 327 54.219 56.643 327 55.237 56.323 328 53.302 57.460 328 52.602 57.853 328 53.255 57.771 328 54.123 59.032 328 53.663 60.311 328 54.026 59.244 328 55.177 60.055 328 51802 58.030 328 51.153 58.692 329 51.225 57.543 329 5S.716 56.924 329 49.867 57.908 329 48.95 56.671 329 94.854 58.614 329 50.64 58.289 330 48.969 59.584 .16A62 -16.349 -15.046 -15.420 -13.843 -13.556 -12.974 -11.821 -10.816 -9.775 -10.071 .649 -12.459 -11.922 -12.604 -13.083 -12.203 -13.135 -12.663 -13.405 -14.81 -14.983 -14.531 -14.522 -15.993 -12.340 -13.094 -11.589 -10.974 -11.610 -13.034 -13.707 -13.224 -14.711 -11.122 -11.488 -10.264 -10.024 -9.608 -9.410 -10.658 -11.016 -11.361 -12.043 -12.380 -12.718 -8.201 .7.588 -7.673 -8.243 -6.266 -5.914 -6.612 4.395 -3.780 -5.976 -6.769 4.873 -4.289 -4.507 4.395 -3.168 -2.296 -2.967 11.37 24.35 21.83 26.39 27A6 0.00 26.27 25.82 23.38 24.43 25.59 25.24 29.36 26.17 26.30 0.00 24.24 19.60 14.81 11.93 20.28 21,10 0.00 0.00 o.co 26.70 25.97 26.25 0.00 23.30 23.42 24.74 24.42 21.44 20.53 24.45 19.21 0.00 18.83 16.54 20.45 17.51 21.65 20.17 18.97 20.53 18.93 17.66 10.15 0.00 16.96 17.42 1949 13.51 25.10 18.38 18.62 20.05 0.00 20.56 17.5 24.14 22.82 22.84 d_~ 4t~" ~ri i. I- L -I L L~ i .i 70 .9 9.

4 SD.

49a0*~ 9 90 a 90 a 4 -9 0s0..

.9 ATOM 1784 H ATOM 1785 CA ATOM 1786 CB ATOM 1781 CG ATOM 1788 CDI ATOM 1789 COZ ATOM 1790 CEI ATOM 1791 CE ATOM 1792 CZ ATOM 1793 C ATOM 1794 0 1795 N ATOM 1796 H ATOM 1797 CA 15 ATOM 1798 CR ATOM 1799 SG ATOM 1800 C ATOM 1801 0 ATOM 1802 N 20 ATOM 1I03 H ATOM 1804 CA ATOM INS5 CB ATOM 2806 00 ATOM 1807 HG ATOM 1808 c ATOM 1809 0 ATOM 1810 N ATOM 1811 it ATOM 1812 CA ATOM 1813 CR ATOM 1814 00 ATOM 1815 110 ATOM 1816 C ATOM 1817 0 ATOM 1118 N ATOM 1119 H ATOM 1820 CA ATOM 1821 CR ATOM 1822 OG1 40 ATOM 1823 HGI ATOM 1824 CG2 ATOM 1825 C ATOM 1826 0 ATOM 1827 N mOm 1828 CD ATOM 1829 CA ATOM 1830 CR ATOM 1131, CG ATOM 1832 C ATOM 133 0 ATOM 8U N ATOM 1835 H ATOM 1836 CA ATOM 1937 CR ATOM 1838 CG ATOM 1839 ODI ATOM 1840 002 ATOM 141 C ATM 1342 0 ATOM 1843 N ATOM 1844 H A*tM 1IM CA ATOM 1846 C ATOM 1847 .CO

PHE

FHE

PHE

PHE

PHE

PHE

PHE

PHE

PHE

PHE

CYS

CYS

CYS

CYS

CYS

CYS

CYS

SER

SER

SER

SER

SER

SPA

SEP.

SEP.

SER

SEP.

SEP.

SER

SER

smt

SER

THR

THR

m

PRO

THR

THR

PRO

PRO

PRO

PRO

PRO

PRO

PRO

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASN

ASN

ASN

ASN

330 48.352 59.323 330 48.369 60.291 330 49.268 61.745 330 49.659 62.459 330 49.771 63.841 330 49.931 61.760 330 50.147 64.514 330 50.309 6.438 330 50.409 63.815 330 47.425 60.183 330 45.536 60.63 331 47.169 59.539 331 47.914 59.170 331 45.831 59.362 331 45.584 57.950 331 45.613 56.747 331 45.649 60.235 331 46.586 60.503 332 44.421 60.692 332 43.659 60.414 332 44.084 61.593 332 42.690 62.162 332 41.835 61.127 332 40.977 61.499 332 44.120 61.144 332 44.290 61.947 333 43.945 59.844 333 43.r,9 59.215 333 43.9V1 59.274 333 42.530 58.895 333 41.566 58.641 333 40.743 51.463 333 44,745 57.979 333 45.104 57.510 334 45.003 57.359 334 4.646 57.711 334 45.720 56.099 334 46.501 56.407 334 47197 55.855 334 48.191 56.183 334 45.753 55.956 334 44.639 54.990 334 43.459 55.273 335 44.853 53.756 335 46.175 53.207 335 43.804 52.736 335 44.56 51.473 335 45.823 52.030 335 42.850 52.545 335 43309 52.608 336 41.545 52.314 336 41.20 52.278 336 40.529 52.137 336 40.940 51.344 336 41344 49.933 336 42.215 49.465 336 40.725 49.302 336 40-046 53.423 336 39.121 53.427 337 40.623 54.569 337 411271 54.602 337 402.24 55.789 '17 Z.L3563 56.352 37 42.065 55.421 -3,694 -1.709 -1.937 -0,647 -0,6332 O-ul 0.21 0.490 1.665 1.653 -1.260 -1.956 -0.117 0.402 0.40-2 0.841 -0.490 1.609 2.342 1.836 1.268 2.923 2.616 2.164 2.030 4.389 5.302 4.657 3.928 5.994 6.23 5.556 6.021 5.19 4.734 6.951 7.785 6.997 8.296 8.169 7.351 9.519 6.904 7.091 6.619 6.414 6.469 6.155 5.529 7.636 8.769 7.406 6.479 8.449 9.710 9.397 10.040 8.531 9.046 9.849 8.686 7.951I 9.372 9.973 1106 1.00 1.00 1.00 .00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 2.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.0 .00 Ioo 100 1.00 .00 100 100 100 I'Ma 1.00 .00 .00 .00 1.00 I.00

LOO

100

LOD

100 .00 1.00 .00 I.00 .00 1.00 1.00 .00 *.00 .00 0.00 18.35 18.09 22.46 21.17 20.68 22.56 20.81 20.07 16.73 20.04 0.00 17.86 16.56 23.49 21.19 20.65 22.22 0.00 21,44 19.68 24.42 0.00 21.99 20,58 23.27 0.00 20.50 21.15 23.77 0.00 23.49 19.23 23.40 0.00 24.09 28.30 34.1! 0.00 14.09 22.20 23.69 20.97 15.82 19.65 20.20 20.66 25.53 29.15 24.20 0.00 28.39 32.12 43.11 52.99 51.49 29.33 34.19 25.69 0.00 23.68 22.73 22.82 j j" bi

_J

71 ATOM 1848 003 ATOM 1349 NIL' ATOM 1850 21 ATOM 1351 H22 ATOM 18&U C ATOM 1853 0 ATOM 1854 N ATOM 1855 H ATOM 186 CA ATOM 187 C8 ATOM 1853 C01 ATOM 1859 CG2 ATOM 1860 C ATOM 1861 0 ATOM 1862 N ATOM 1863 H ATOM 1364 CA ATOM 1865 CD ATOM 1366 00 ATOM 1367 HG ATOM 1368 C ATOM 1569 0 ~ATOM 18M N ATOM 1871 H 25 ATOM 1372 CA ATOM 1873 CD ATOM 1374 CO ATOM 1875 002 ATOM 1876 CE2 ATOM 1377 03 ATOM 187 CDI ATOM 1879 NEI a ATOM 1881 EI ATOM 182 02 ATOM 1883 0 C02 ATOM 188 C ATOM IM 0 ATOM IU N ATOM 187 H ATOM 1888 CA ATOM 189 CB ATOM I890 CO ATOM 1891 CD 45 ATOM 192 NE ATOM 1893 HE ATOM 1894 CZ ATOM 1895 NHI ATOM I896' H113 ATOM 3897 12 ATOM I898 NH2 ATOM 3899 HH21 ATOM 1900 HH2 ATOM 190! C ATOM 1902 0 ATOM 1903 N ATOM 3904 H ATOM 3905 CA ATOM 2906 CD ATOM 907 CG ATOM 19W8 CD2 ATOM 3909 NDI ATOM 1910 HID1 ATOM 1913 CS)

ASN

ASH

ASN

ASN

ASN

ASN

VAL

VAL

VAL

V'AL

VAL

VAL

VAL

VAL

SER

SER

SEIR

551

SEP

SER

551

SER

SER

TRP

TRP

TRP

TRP

TRP

TIP

TRP

TRP

TRP

YRP

TRP

TRP

ARO

ARO

ARO

ARG

ARG

ARO

APG ARG

ARG

ARG

ARO

ARG

ARO

ARG

ARG

AIS

ARG

IS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

337 41379 54.991 337 43193 54.940 337 43.322 55.204 337 43.604 54.320 337 39.600 56.312 337 39.737 56.360 333 38.815 37.691 338 33.760 57.662 333 38,117 58.722 338 36.902 59.297 3398 35.901 58.187 338 37.369 59.946 338 39.045 59.870 333 40.124 60.040 339 33,588 60.680 339 37.756 60.450 339 39.231 61.948 339 39.666 62.030 339 39.374 60.771 339 39.097 60.209 339 38.381 63.015 339 37.008 62.669 340 38.549 64.280 340 39.494 64.546 340 37.530 65.306 340 38.110 66.071 340 38.069 65.153 340 36.70 64.901 340 37.231 63.820 340 35.473 65.302 340 39.112 64.394 340 38.611 63.616 340 39.143 62.912 340 36.364 63.217 340 34.612 64.660 340 35.043 63.639 340 37.260 66.265 340 38.137 66.652 341 35.989 66.670 34t 35.321 66.289 341 35.487 67.607 341 34.617 66.813 343 34.391 67.533 341 33.517 66.655 341 32.151 66.768 341 31.684 67.621 341 31.501 65.767 341 32.089 64.556 341 33.030 64.400 341 31.577 63.82! 343 30.219 65.962 341 29.717 66.448 341 29.720 65.218 343 3A.616 68.68 341 33,969 68.486 342 34.550 69.880 342 35.002 70.055 341 33.820 70.96% 342 34.85 "1.921 34, 34.28 73.095 342 33.508 74.035 342 34.247 73.368 342 3466t 72.34 34? 33.458 74.387 11.997 10.919 10.231 11.691 .496 7.283 9.095 lo~on 11 9.169 9.486 10.467 8.024 3.588 7.067 6.$96 6.743 5.285 4.672 4.677 6.948 6.924 7J130 7.074 7.447 3,665 91516 11.450 10.320 11.256 11.687 12.277 11.38 12.227 6,311 5.544 6.210 6.323 5.211 4.170 2.361 1.972 2.441 2.324 3.031 3.232 2.934 3.676 3.440 3.294 3.814 5.864 6.883 5.324 4.473 5.934 6.463 7.179 6.600 9.511 9.249 3.7m3 3.00 1.00 1.00 1.00 1.00 1.00

LODO

1.00 1.00 I.00 1.00 .00 L~oo 1.00 1.00 3.00 .00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 .00 1.00 3.oo 1.00 3.00 1.00 1.00 3.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 I3.00 I.00 1.00 1.00 1.00 1.00 1.00 I bo 1.00 1.00 1.00 1.00 LODo 25.91 38.06 0.00 0.00 25.83 29.96 27.31 0.00 24.45 24.69 25.15 25.43 25.97 26.13 27.49 0.00 32.77 30.77 38.35 0.00 32.35 29.99 31.13 0.00 32.42 28.07 3.22 33.69 38.45 37.14 32.03 35.26 0.00 43.31 42.02 45.40 35.22 36,37 37.03 0.00 36.96 34.44 37.33 39.02 48.98 0.00 51.03 56.13 0.00 0.00 U3.22 0.00 0.00 37.23 38.6 39.36 0.00 43.08 41.90 45.95 46.70 43.33 0.00 48.44 72 ATOM 1912 NEM HIS ATOM 1913 112 HIS ATOM 1914 C HIS ATOM 1915 0 HiS S ATOM 1916 N PRO ATOM 1917 Co PRO ATOM 1918 CA PRO ATOM 1919 Ca PRO ATOM 1920 CO PRO ATOM 1921 c PRO ATO16 192-1 0 PRO ATOM 1923 N THR ATOM 1924 H THR ATOM 9 CA THR 15 ATOM 192- CB THR 41 ATOM 1927 001 THR ATOMI me Hr0 THR ATOM 1929 CG2 THR ATOM 1930 C 7HR ATOM I9i TH **lees ATOM 13 N MET ATOM 1933 1 MET ATOM 1934 CA MET CATOM M35 CB ME ATOM 1936 CG ME ATOM 1937 5D MET ATOM 1938 CE MuT ATOM 1939 C MET ATOM 1WO 0 MET ATOM 94 N (3LY ATOM 19C H SILY ATOM 1943' CA GLY ATOM 1944 C GLY ATOM 194S 0 GLY 35 ATom 1m N SEll s ATOM 1947 1R Sml ATOM 198 CA SER ATOM 1949 C SER ATOM 1950 00 SEl ATOM 1951 1( SEP ATOM 1952 C _SEl ATOM 1953 0 SE ATOM 1954 N VAL ATOM 1955 11 VAL ATOM 1956 CA VAL ATOM 195 Co VAL ATOM 1958 CGI VAL ATOM 1959. C2 VAL ATOM 1960 C VAL ATOM 1961 0 VAL ATOM 1962 N PHE ATOM 163 H PHE ATOM 1964 CA PHE ATOM 1965 CS PHIE ATOM CG66 CO pmlE ATOM 1967 CDI PPE ATOM 1963 CD2 PHE ATOM 1969 CE PlE ATOM 19710 CE2 PHE ATOM 1971 Cz PHE ATOM 1972 C PtlE ATOM 1973 0. PHE ATOM 1974 N M1E ATOM 1975 H nm 342 33.020 74.776 7.42 342 32.497 75.591 7.407 342 32.394 71.612 4.99 342 33.367 71.960 3.826 343 31.637 71.344 5.101 343 30.934 71.752 6.98 343 30,678 72.2W .073 343 29.317 72.590 4.824 343 29.512 71.689 6.015 343 31068 73.489 3.199 343 30.05 73.535 1.88 34t 31.615 74.459 3.909 344 31.830 74.264 4.837 344 31.911 75.71 3.334 344 31.577 6.802 4.429 344 31.901 76.175 5.672 344 31.164 75.546 5.7S2 344 30.08 77.164 4.490 344 33.367 73.826 2.944 344 33.82 76.516 2.046 345 34.192 75.055 3.632 345 33.344 74.348 4.

345 35.604 75.172 3.363 345 36.357 7S.29 4.662 345 36.128 76.474 5.498 345 36.752 76.173 7.T76 345 38.40$ 76.807 7.023 345 36.133 74.006 2.553 345 37.28 4.042 2.179 346 35383 72.947 2.236 346 34.439 72.392 2.503 346 .35.91 71.222 1.544 346 -36.764 70.915 2.495 346 36W 70.988 3.718 347 37.567 70.012 1.949 347 37.693 70.023 0.990 347 38.274 68.999 2.69 347 38.434 67.768 1.820 347 37.2 67.491 1.098 347 36.547 67.238 1.697 347 39.631 69.491 3.157 347 40.397 70.136 2438 344 39.580 69.144 4411 38 39.30 68343 4.87 348 41127 69583 5.086 348 41.215 68,976 6.494 343 42237 69.746 1.309 348 39.895 69.060 7.219 34 42.345 69.177 4.305 343 43.247 69.963 4.071 349 42.425 67.917 3.362 349 41.695 67.278 4.013 349 43,621 W7S5 3.181 349 41443 65.960 2.908 349 44.459 65.361 1.941 349 44.029 64.916 0.702 349 45.70 65223 2.311 349 4.919 64.273 .121 349 46.677 64590 1.459 349 46.232 64.101 0.25 349 43.84 63.243 1.89 349 44.966 68.632 1609 350 .42.7 68.499 1.097 3m0 41.912 68.20 .3m6 1.00 .00 1.0 1.00 .00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 3.00 I.00 I.00 1.00 1.00 1.00 1.00 1.00 1.0 1.00 1.00 1.00 1.00 1.00 1.00 _.00 1.00 I.00j 1.00 1.00 1.0 1.00 1.00 I.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 oo 1.00 100 1.00 3.00 1.00 1.00 3.00 3.00 1.00 1.00 1.00 1.00 I Vo 1.00 1.00 1.00 49.68 0.00 46.13 43.57 48.28 46.23 49.93 44.40 41.79 33.21 35.64 54.13 0.00 53.54 56.09 63.15 0.00 57.44 53.06 53.90 51.25 0.00 52.69 60.31 71.30 86.57 81.62 47,27 48.63 40.77 0.00 35.53 40.26 32.41 0.00 28.45 24.33 30.64 0.00 -31.86 33.18 29.14 0.00 25.67 27.65 29.29 29.60 25.79 33.26 23.67 0 00 22.69 38.76 20-55 4.,77 35.53 1644 13.87 12.07 2364 23.92 23.51 0 QQ 000 -I 73 ATOM 1976 CA SATOM 1977 CBa ATOM 1978 CG2 ATOM 1979 CGI ATOM 1980 CDI ATOM 1981 C ATOM 1982 O0 ATOM 1983 N ATOM 1984 Hi ATOM 1985 CA ATOM 1986 C ATOM 1987 O0 ATOM 1988 N ATOM 1989 1 ATOM 1990 CA ATOM 1991 CB ATOM 1992 CG ATM 1993 CD ATOM 1994. NE 20 ATOM 1995 HE ATOM 1996 CZ ATOM 1997 NHI ATOM 1998 HHI ATOM 1999 H111112 ATOM 2000 NH2 ATOM 2001 HH21 ATOM 2002 111122 ATOM 2003 C ATOM 2004 0 ATOM 2005 N ATOM 2006 H ATOM 2007 CA ATOM 2008 CB SATOM 2009 CG S 35 ATOM 2010o CDi ATOM 2011 CD2 ATOM 2012 C ATOM 2013 0 ATOM 2014 N ATOM 2015 H ATOM 2016 CA ATOM 2017 CB ATOM 2018 C0G2 ATOM 2019 CGI ATOM 2020 CD1I ATOM 2021 C ATOM 2022 O0 ATOM 2023. N ATOM 2024 11 ATOM 2025 CA ATOM 2026 CB ATOM 2027 CG ATOM 2028 CD ATOM 2029 OEI ATOM 2030 OE21 ATOM 2031 C ATOM 2032 O0 ATOM 2033 N ATOM 2034 It GO ATOM 2035 CA ATOM 2036 CB ATOM 2037 CG ATOM 2038 CD2 ATOM 2039 NDI 350 42,969 69.170 350 41,617 69.131 350 41.741 69.922 350 41.200 67,689 350 42,089 66.972 350 43.451 70,600 350 44.428 71,058 351 42.813 71,359 351 42.057 70,988 351 43.248 72,720 351 44.702 72.800 351 45.524 73.568 352 45.064 71,963 352 44,388 71.391 352 46.449 71.872 352 46.589 70.842 352 47,697 71.286 352 47.091 71.997 352 47.179 71.107 352 47.920 70.468 352 46.307 71.116 352 46.502 70,272 352 47.294 69.660 352 45.849 70.267 352 45.227 71.943 352 45.080 72,554 352 44,570 71.936 352 47.419 71.483 352 48.535 71.995 353 47,033 70.567 353 46.156 70.135 353 47.866 70.154 353 47.222 68.947 353 47,975 68.452 353 49.420 68.121 353 47.258 67,230 353 48.034 71.303 353 49.142 71.600 354 46,923 71.977 354 46.049 71.660 354 47,019 73,174 354 45,595 73,782 354 45.652 75.176 354 44,769 72,893 354 43.279 73.267 354 48.000 74.172 354 48.967 74.616 355 47.806 74.559 355 47.040 74.212 35$ 48.719 75.507 355 48.501 75.685 355 47,165 76.212 355 47.053 75.777 355 45.921 75.683 355 48.092 75.513 355 50.150 75.114 355 51,029 75.893 356 50.461 73.862 356 49,739 73.225 356 51.855 73.438 356 51.997 72.019 356 51.846 72.065 356 52.890 72.264 356 50.728 72.044 -0.185 -0.939 -2.229 -1,267 -2.272 0.073 -0.510 0.962 1.467 1,212 1.640 1,138 2.614 3.045 3.042 4.167 5.104 6.310 7.452 7.491 8.449 9.494 9.507 10.251 8.435 7,657 9,187 1.931 1.866 1.037 1.143 -0,070 -0,778 -2.020 -1.696 -2,552 -1.031 -1,485 -1,350 -1.035 -2.168 -2.327 -2.940 -3.248 -3.187 -1.540 -2.165 -0.276 0,235 0,341 1.803 2.282 3.746 4.245 4.385 0.242 0054 0.489 0.663 0.432 1.041 2,551 3.421 3.288 23.11 23,.98 24.02 20.37 14.33 23.87 25.61 21.19 0.00 19,54 26.25 28.41 28.53 0,00 26.92 31.79 35.13 38,95 44.61 0,00 41.51 48,56 0.00 0.00 44,65 0.00 0.00 26.87 26.27 25.01 0.00 23,26 25.55 23.59 25.86 29.54 27,31 24.83 22.68 0.00 24,25 21.11 21,96 17.07 18.04 25,12 27.26 27.20 0.00 25.06 3273 45,70 58.26 61.90 61,89 23,.42 25.26 26.55 0.00 26.58 27.11 23 74 22.80 25.98

A

pp~ rri ru- I r r~i- l- 111-- ~1 X~ 74 ATOM 2040 HDI HIS ATOM 2041 CEI HIS ATOM 2042 NE2 HIS ATOM 2043 HE2 HIS ATOM 2044 C HIS ATOM 2045 0 HI ATOM 2046 N MET ATOM 2047 H MET ATOM 2048 CA MET ATOM 2049 CB MET ATOM 2050 CG MET ATOM 2051 SD MET ATOM 2052 CE MET ATOM 2053 C MET 5 ATOM 2054 0 MET ATOM 2055 N GLN ATOM 2056 H GLN ATOM 2057 CA GLN ATOM 2058 CB GLN 20 ATOM 2059 CO GLN ATOM 2060 CD GLN ATOM 2061 OEL OLN ATOM 2062 NE2 GLN ATOM 2063 HE1I GOLN ATOM 2064 HE22 GLN ATOM 2065 C GLN ATOM 2066 O GLN ATOM 2067 N GLU ATOM 2068 H GLU ATOM 2069 CA GLU ATOM 2070 CB GLU ATOM 2071 CO OLU ATOM 2072 CD GLU SATOM 2073 OE GLU 35 ATOM 2074 OE2 GLU ATOM 2075 C GLU ATOM 2076 O GLU ATOM 2077 N TYR ATOM 2078 H TYR 40 ATOM 2079 CA TYR ATOM 2080 CB TYR ATOM 2081 CO TYR SATOM 2082 CDI TYR ATOM 2083 CEI TYR 45 ATOM 2084 CD2 TYR ATOM 2085 CE2 TYR ATOM 2086 CZ TYR ATOM 20817. OH TYR ATOM 2088 11 TYR SO ATOM 2089 C TYR ATOM 2090 0 TYR ATOM 2091 N ALA ATOM 2092 H ALA ATOM 2093 CA ALA ATOM 2094 CR ALA ATOM 2095 C ALA ATOM 2096 O ALA ATOM 2097 N CYS ATOM 2098 H CYS ATOM 2099 CA CYS ATOM 2100 C CYS ATOM 2101 0 CYS ATOM 2102 CB CYS ATOM 2103 SG CYS 356 49.819 71,907 356 51.036 72.231 356 52.333 72.360 356 52.814 72,450 356 52.411 73.447 356 53.560 73.809 357 51.586 73.039 357 50.672 72732 357 52.037 73.060 357 50.995 72.413 357 50.860 70.870 357 52.182 69.919 357 51.449 69,433 357 52.249 74.503 357 53.237 74,846 358 51.365 75,428 358 50.339 75.168 358 51.650 76,808 358 50.505 77693 358 49,262 77,438 358 48,063 78,234 358 47,170 78.570 358 47,919 78.558 358 48,611 78.285 358 47.124 79.077 358 52,982 77,344 358 53,783 77,971 359 53,234 77.083 359 52.581 76.563 359 54,425 77.55& 359 54.178 77.382 359 55.215 77,874 359 55.241. 79,379 359 54.197 79,992 359 56.320 79.940 359 5.638 76,809 359 56.664 77422 360 55.584 75.475 360 54.723 75,002 360 56.814 74,710 360 56.902 73.528 360 56.910 74,022 360 55.765 73,873 360 55.711 74,433 360 58.022 74.677 360 57.974 75.239 360 56.813 75.114 360 56.737 75.692 360 55.852 75.603 360 57,148 74.120 360 58,243 73.583 361 56,288 74.169 361 55.459 74.682 361 56.544 73.494 36. 55,328 73.717 361 57,818 73.954 361 58,461 73.249 362 58,273 75.178 362 57.694 75.795 36Z 59,576 75.578 362 60.768 74.919 362 61,761 74,329 362 59.746 77,086 362 61.400 77.655 2.953 4.550 4,608 5,460 -0.983 -1.239 .1,948 .13.747 -3316 -4239 4.254 5.054 -6.584 -3,704 -4.338 -3,360 -2,897 -3709 -3.214 4.067 -3.596 4362 -2.329 -1.693 -1.095 -3,182 -3.869 -1,906 -1.386 1243 0.251 1.245 1,308 1.550 1.332 -13721 -1.974 -1.867 -1,843 -2,053 -1.079 0.333 1.087 2.342 0.82.4 2.080 2,090 2,823 4.073 4,430 3.389 3523 -4.411 -4320 -$.678 -6.600 -6,355 4,116 -6,133 .d5620 -6.659 -5.987 -6.575 -6.546 -7.050 0.00 19,00 22,63 0,00 26.45 20.78 17.38 0,00 28.28 30,75 34.53 4328 37,66 29.20 31.64 25.95 0,00 27.58 24,97 26.18 28.54 30.13 31.54 0,00 0.00 30.98 31,45 32.65 0.00 32.96 37.33 S1,69 63.61 70,65 69,55 33.19 36,20 33.15 0.00 28,52 30.16 28,83 30.92 33,76 30,38 32,36 35.09 41375 0.00 27.85 27.93 27.66 0.00 28.82 26.34 31.26 35.01 38.50 0.00r 42.41 41.51 41.46 47.61 64.18 (C99 75 ATOM 2104 N ATOM 2105 11 ATOM 2106 CA ATOM 2107 CD ATOM 2108 OG ATOM 2109 HG ATOM 2110 C ATOM 2111 0 ATOM 2112 N ATOM Zt13 11 ATOM 2104 CA ATOM 2115 Ca ATOM 2116 So ATOM 2117 C ATOM 2118 0 ATOM 2119 N ATOM 2120 11 ATOM 2121 CA ATOM 2122. CB ATOM 2123 CG ATOMf 2124 CDI ATOM 2125 OD2 ATOM 2126 C ATOM 2127 0 ATOM 2128 N ATOM 2129 H ATOM 2130 CA ATOM 2131 CB ATOM 2132 Cot ATOM 2133 CG2 ATOM 2134 C ATOM 2135 0 ATOM 2136 N4 ATOM 2137 H 35 ATOM 2138 CA ATOM 2139 CR ATOMN 2140 CG ATOM 2141 L CD ATOM 2142 O13 ATOM 2143 OE ATOM 2144 C ATOM 2145 0 ATOM 2146 N ATOM 2147 H ATOM 2148 CA ATOM 2149 CB ATOM 2150 CG ATOM 2151. CD ATOM 2152 OEI ATOM 2153 0E2 ATOM 214 C ATOM 2153 0 ATOM 2156 H ATOM 2157 1 ATOM 2158 CA ATOM 2159 CB ATOM 2160 CG2 ATOM 2161 cot ATOM 2162 CDI ATOM 2163 C ATOM 2164 0 ATOM 2165 N ATOM 2166 H ATOM 2167 CA

SER

SER

SER

SER

SER

SER

SEl

SER

CYS

CYS

CYS

CYS

CYS

CYS

CYS

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

ASP

VAL

VAL

VAL

VAL

VAL

VAL.

VAL

VAL

GLU

OLU

CLU

GLU

GLI

GLU

GLO

GLU

CLI

CLV

GLU

GLU

GLU

GLU

GLI

CLI

GLI

GLU

GLU

IU

ILE

ILE

ELE

ILE

ILE

ILE

ILE

ILE

PHE

PRE

PRE

363 60,741 74.757 363 59.909 74.803 363 61,955 74.338 363 62.122 75.236 363 60.899 75.837 363 60.553 76,414 363 61.985 72186 363 63,036 72.255 364 60,857 72.220 364 59.916 72.633 364 60.937 70.839 364 60,056 70.623 364 60,508 71.767 364 60,479 69.967 364 59.610 70,34 365 61.025 68.770 365 61.859 68.579 365 60.406 67.784 365 61.379 66.619 365 61.757 65.825 365 61.026 64,918 365 62.805 66.094 365 59.111 67.254 365 58.838 67420 366 58.277 66.590 366 58.54 66,434 366 56.968 66,183 366 56.251 65.513 366 55.102 64.601 366 55,629 66.654 366 57,007 65.309 366 56,256 65.536 367 57.856 64.298 367 58.450 64,111 367 57.867 63.516 367 58.939 62.428 367 58.553 61.219 367 59,442 60.019 367 59,643 59.736 367 59.937 59,388 367 58.106 64,352 367 57.604 64.052 368 58.872 65,437 368 59,242 65.125 368 59,094 66231 36& 60.278 67,145 369 61.017 66,888 369 61.471 68.195 368 61.848 69.099 368 b6.444 68.298 368 57,923 67.082 368 57.561 67.224 369 57.268 67,677 369 7.595 67.596 369 56.027 68.395 369 55.403 68.931 369 54.089 69.671 369 56.360 69.917 369 55.911 70.256 369 55.024 67.497 369 54.331 67,885 370 54,923 66.241 370 55470 65.932 370 53.962 65.359 -4.681 .4165 -4.026 -2.80') -2.362 -3,046 -3,661 -3,593 3,399 -3.542 -2,979 -1.755 -0.422 4.121 -4,887 4.300 -1828 -5,164 .5,4 19 4171 -3,783 -3,599 .,538 -3.343 -5,336 -6.268 857 -6,051 -5.672 .6,848 -3.611 -L674 -3.489 4.237 -2,259 -2.289 -3.144 -2.820 -1.632 -3.767 -1.012 0,0519 -1.959 0.076 -0,069 1,244 1.41 1,078 3.071 0.396 1.55$ -9,59 -1.5 13 -0.295 .1,624 -1,355 -2.2U 3701 0.435 1.371 0.020 -0.733 0.654 1.00 1.00 1,00 1.00 I0 1.00 1.00 1,00 1.00 1.00 1,00 1.00 1,00 1,00 1,00 1.00 1.00 1.00 1.00 1.00 1100 1,00 1.0 1.0 .0D 1.00 .00 .00 1.00 I .00 1,00 1.00 I-Q0 1,00 1.00 .00 .00 1.00 1.00 .00 .00 1,00 1,00 1.0 1.00 ,00 1.0 1.00 1.00 1.00 1,00 1.00 .00 .00 1.00 1.00 1.00 1.00 40.95 0.00 43.35 44,65 53.93 0.00 40.77 43,37 3.31 0.00 33,39 32.55 40.02 29,84 30,07 28.01 0.00 27.07 27.73 31.83 34.71 42,02 28.95 27,98 27.55 0.00 27.84 24.13 23.78 23.21 28.22 30.81 28.49 0.00 27.85 26.64 28.69 37.41 42.87 37,88 28.61 31,13 28,39 0.00 26.65 34.26 53.59 63.52 71.58 64,.40 23.42 27.18 22.22 0.00 19.89 22.54 20.28 17.77 15.38 20.18 20.19 21.39 0.00 23,30 rn 76 ATOM 2168 CB PRE ATOM 2169 CG PRE ATOM 2170 CDI PRE ATOM 2171 CD2 PHE ATOM 2172 CEI PRE ATOM 2173 CE2 PRE ATOM 2174 C2 PHE ATOM 2175 C PHE ATOM 2176 0 PlE ATOM 2177 N ARG ATOM 2178 H ARG ATOM. 2179 CA ARG ATOM 21&0 CB ARG ATOM 2181 CG ARO ATOM 2)82 CD ARG ATOM 2183 NE ARO ATOM 2184 HE ARG ATOM 2185 CZ ARG ATOM 2186 NlH) A1G ATOM 2187 HII ARO ATOM 2188 HIZ ARG ATOM 2189 N112 ARO ATOM 2190 H12 ARG ATOM 2191 HH22 ARO AToM 2192 c ARG ATOM 2193 0 ARG ATOM 2194 N LYS ATOM 2195 H LYS ATOM 2196 CA LYS ATOM 2197 CB LYS ATOM 2198 CG LYS ATOM 2199 CD LYS ATOM 2200 CE LYS ATOM 2201 NZ LYS 35 ATOM 2202 HZI LYS ATOM 2203 HZ2 LYS ATOM 2204 HZ3 LYS ATOM 2205 C LYS ATOM 2206 0 LYS ATOM 2207 N VAL ATOM 2209 11 VAL ATOM 2209 CA VAL r. ATOM 2210 CB VAL ATOM 211 Co VAL ATOM 2212 C02 VAL ATOM 2213 C VA.

ATOM 2214 0 VA, ATOM 2215 N ARG ATOM 2216 H ARG ATOM 2217 CA AR..

ATOM 2218 CB ARG ATOM 2219 CO ARG ATOM 2220 CD ARCI ATOM 2221 NE ARO ATOM 2222 HE ARG ATOM 2223 CZ ARG ATOM 2224 NHI ARG ATOM 222.5 R11 ARG ATOM 2226 HU12 ARO ATOM 2227 11N12 ARG ATOM 2228 HH21 ARG ATOM 2229 1122 ARO ATOM 2230 C ARG ATOM 2231 0 ARG 53.842 64.029 53140 64.222 3,037 63.153 52.624 65,444 52.427 63.311 32,013 65,595 51.96 64.521 54350 65,076 53.5.3 65.031 55.634 64,883 56.291 64804 56.103 64,783 57,611 64.627 57.954 64.338 59.453 64.358 60,022 65.650 60.521 65,766 59.899 66,694 59207 66,648 58.750 65.80 59.163 67.463 60.525 67.849 61.067 67.884 60.471 68,649 55.704 65.999 53.105 65.907 56,020 67.200 56,525 67.281 55,576 68.377 56.008 69,609 57.506 69.718 57,933 70.852 59,042 71.66 58.499 72.323 58.138 71.612 57,717 72,944 59.239 72,893 54.083 68.434 53617 68,867 53,240 68.015 53.577 67,7123 51.813 67977 50.922 67.520 49.A42 67.520 31.108 69,476 51.588 67,006 50.792 67.332 52.251 65,839 52.897 65,603 52,023 64,926 52.827 63.601 52.332 62.777 52.822 61.333 54.262 61.249 54,727 61.264 54,973 6.1.142 54333 61.136 53.340 61.217 54.864 61.061 56.344 61.046 56.811 61.053 56,874 60.96, 52423 65,570 51.724 63.539 -0.121 .1,Z1.

-2.304 -1.841 -3.533 -3,069 -3,919 2,069 2.978 2,314 1.573 3,683 3,722 5.180 5.411 5.122 4.286 1.969 7,152 7,435 7.729 5.632 4.793 6.231 4.50 5.576 4.020 3,182 4.733 3.976 4.053 3.136 3.89 4.969 5,636 4,674 5.426 4.960 6.008 4,026 3.154 4,333 3.121 3.541 1.933 5.471 6,336 3.540 4.838 6,660 6,511 5,307 5.253 5.278 6,141 4.161 2.923 2.126 4.208 5A094 3.362 &.958 8.933 23.30 23,92 27.0 29.88 26,34 27,67 27.89 23.38 24.64 24.62 0,00 26.21 25.42 26.24 27.46 34.21 0.00 40.22 39,64 0,00 0,00 39.43 0.00 0.00 29,69 32,42 26.46 0.00 23,67 25.18 21.18 33.26 42.84 50.90 0,00 0.00 0,00 22,44 25.03 24.85 0.00 26-14 29.33 23.06 20,17 26.25 25.40 22.91 0.00 24.92 24.92 26.50 21.69 21.59 0.00 19.13 22.36 0.00 0.00 22.93 0.00 0.00 23.27 20.79 4.

il^riua~- 77 ATOM 2232 N ATOM 2233 11 ATOM 2234 CA ATOM 2235 ca ATOM 2236 CG ATOM 2237 CDI ATOM 2233 CD2 ATOM 2239 CEI ATOM 2240 CE2 ATOM 2241 CZ ATOM 2242 C ATOM 2233 0 ATOM 2244 N ATOM 2245 H ATOM 2246 CA ATOM 2247 CB ATOM 2248 OG ATOM 2249 HO ATOM 2250- C ATOM 2251 0 ATOM 2252 N ATOM 2253 H ATOM 2254 CA ATOM 2255 CB 25 ATOM 2256 C ATOM 2257 CDI ATOM 223 CDI ATOM 2259 CEI ATOM 2260 CEZ ATOM 2261 CZ ATOM 2262 C ATOM 2263 0 ATOM 2264 1 ATOM 2265 H 35 ATOM 2266 cA ATOM 2267 CB ATOM 2263 CG ATOM 2269 CD ATOM 2270 OEi ATOM 2271 0hZ ATOM 72 C ATOM 2273 0 ATOM 2274 N *ATOM 2275 H ATOM .76 CA ATOM 2277 CR ATOM 2278 CO ATOM 2279. CD ATOM 2280 OEI ATOM 2281 NE2 ATOIN 2282 HEN2 ATOM 223 HME2 ATOM 2284 C ATOM 2283 0 ATOM 2286 N ATOM 2237 C ATOM 2288 CA ATOM 2239 CR ATOM 2290 Cc ATOM 2291 C ATOM 2292 0 ATOM 2293 N ATOM 2294 11 ATOM 2295 CA 375 53.581 66.204 375 54,147 66.181 375 54,025 66.896 375 55.388 67.523 375 56,010 68.272 375 56.926 67,655 375 55-684 69.589 375 57.513 68.348 375 56.291 70.277 375 57.202 69.659 375 52.982 67.917 375 52.775 68,13 376 52.260 68.589 376 52,414 68.468 376 51,291 69,562 376 50,715 70.398 376 49.984 69.740 376 50.480 68.972 376 50,124 68.979 376 49.395 69,683 377 49,927 67,671 377 50.511 67127 377 48,892. 66.990 377 48.276 65,883 377 47.359 66,445 377 47.815 66,654 377 46,00 66.714 377 46.933 67.097 377 45,173 67,155 377 45613 67.337 377 49.407 66.381 377 48.713 65,615 378 50.634 66.675 378 51151 67.389 378 51,169 65.910 378 52,649 66.192 373 53.146 64,886 378 54,632 64,790 373 55130 63.668 378 55.292 65.826 378 50.502 66.092 378 50.366 65.153 379 50.053 67.255 379 50J449 68039 379 49.268 67.367 379 49.380 6,7838 379 50.771 69.437 379 511711 68.603 379 51.480 68.345 379 52.809 68,101 379 52.996 68.731 379 73.71 67.564 379 47.797 67,066 379 47.241 67.723 380 47.099 66157 380 47,634 65.095 380 45,670 65,984 380 45.261 64.315 380 46.548 64.024 380 44.890 67.142 380 44.736 67.733 31 44.366 67.794 381 44.538 67.356 331 43.596 69.021 7,996 7,193 9,187 8,828 9989 10.801 10.221 11,838 11.255 12.064 9.664 10.857 8,742 7,783 9.208 8,047 7.034 6.709 9.960 10.647 9,838 9.265 10.570 9,691 8,647 7.363 8961 6.390 7.981 6.638 11 860 12.528 12.279 11.850 13.374 13.534 14.168 14.234 14.315 14.210 14,719 15,483 15.116 14.543 16.328 16,896 16,941 17,767 13.938 17.225 16.281 17.826 16,009 15.130 16602 17.439 16.367 17.243 17.310 16.664 17.776 15.586 14.721 15,617 24.10 0.00 23.34 29.16 35.30 36.99 33.65 36,47 38.44 37.61 28.21 28.91 28.48 0.00 28,11 24.30 26.36 0.00 30.03 27.69 33.33 0.00 30.24 21.45 21,79 23.05 22.87 21.35 16.45 30,42 29,72 30,87 0.00 35.50 33.01 3.65 42.24 45,42 51,72 36.27 39.55 39.19 0.00 48.22 56.51 63.32 69.92 74,89 72.82 0.00 0.00 50.39 51.95 51.98 50.16 58.10 53.72 49.03 67.86 70.34 77.19 0.00 83.19 -nc mc~i~,-xMn~ntli~~ ~nrp~_i 78 r i t i t i.

ATOM 2296 CB ATOM 2297 CG ATOM 2298 ODI ATOM 2299 OD2 ATOM 2300 C ATOM 2301 0 ATOM 2302 N ATOM 2303 H ATOM 2304 CA 10 ATOM 2305 C ATOM 2306 O0 ATOM 2307 N ATOM 2308 H ATOM 2309 CA ATOM 2310 CU ATOM 2311 CG ATOM 2312 CD ATOM 2313 NE ATOM 2314 HE ATOM 2315 CZ ATOM 2316 NHI ATOM 2317 HH11 ATOM 2318 HU12 ATOM 2319 H112 25 ATOM 2320 1 ATOM 2321 H1122 ATOM 2322 C ATOM 2323 0 ATOM 2324 N ATOM 2325 H ATOM 2326 CA ATOM 2327 CB ATOM 2328 C X. ATOM 2329 0 ATOM 2330 N ATOM 2331 H ATOM 2332 CA ATOM 2333 CE ATOM 2334 CG 40 ATOM 2335 CD ATOM 2336 OE ATOM 2337 Nt2 ATOM 2338 HE21 ATOM 2339 HE22 ATOM 2340 C ATOM 2341 0 ATOM 2342 N ATOM 23431 ATOM 2344 CA ATOM 2345 CA ATOM 2346 CU ATOM 2347 SD ATOM 2348 CE ATOM 2349 C ATOM 2350 0 ATOM 2351 N ATOM 2352. CD ATOM 23W3 CA ATOM 2354 CB ATOM 2355 CG ATOM 2356 C ATOM 237 0 ATOM 238 N ATOM 2359 H

ASP

ASP

ASP

ASP

AS?

ASP

GLY

GLY

GLY

GLY

GLY

ARG

ARG

ARG

ARG

ARG

ARG

ARG

ARGC

ARG

ARG

ARG

ARG

ARC

ARG

ARC

MA

ARG

ARG

ALA

ALA

ALA

ALA

ALA

A

GLN

GLN

GLN

GLN

GLN

GL

GLI

GLN

GLN

CILN

GLN

MFT

MET

MET

MET

MET

MET

MET

MR

MET

PRO

PRO

PRO

PRO

PRO

PRO

PRO

THR

THR

381 43.541 69.494 381 43,014 70,910 381 43.143 71.746 381 42481 71.167 381 42.223 68.750 381 41,197 69.227 382 42.096 67.981 382 42.906 67,764 382 40.780 67.478 382 40.321 66448 382 40.458 65.247 383 39.759 66,873 383 39.519 67,824 383 39.629 66,003 383 39.071 66,738 383 37.832 67.576 383 36,696 66741 383 35.525 67.573 383 35.556 68.516 383 34.408 67.087 383 33.341 67.925 383 33.413 68.873 383 32.487 67.594 383 34.322 65,793 383 35.108 65.180 383 33.469 65.461 383 41.041 65.557 383 41.896 66.385 384 41.392 64.279 384 40.809 63.578 384 42,711 63,902 384 43.392 63,101 384 42.614 63,088 384 41.666 62.329 385 43.604 63.245 385 44.264 63.968 385 43.794 62.325 385 42.964 62733 385 43.318 64,098 385 42.355 64,422 385 41.375 65.110 385 42598 63,875 385 43.370 63.272 385 42.007 614105 385 45.276 62.3904 385 45,955 63312 386 45.844 61.457 386 45253 60,770 386 47.2$1 61.417 386 47.980 60.448 386 49.459 60.203 386 50.369 61.710 386 50.297 62.580 386 47,376 60.901 386 47.187 59.706 387 47.623 6.10 387 47,479 63,144 387 47,706 61.309 387 47.813 62609 387 48.153 63.635 387 48.825 60.365 387 49.849 60.377 388 48.671 59.504 338 47.904 59.580 14. 153 13.938 14,643 12.846 16.235 15,766 17,316 17.831 17.697 16.669 16.841 15.545 15.480 14.381 13-173 13.414 13.993 14,194 13.929 14732 14,847 14.534 15.250 15,173 15.101 15,576 14.005 13.710 13.985 14.357 13.518 14,618 12.237 12,.029 11.349 11,422 10.258 9.034 8,481 7.399 7.586 6.223 6,110 5,478 9.937 10.363 9.197 8,827 8,921 9.881 9,609 9.227 10.768 7503 7.306 6.539 6.670 5.'134 4.355 5.396 4.553 5.523 3.859 3.255 1,00 1.00 1.00 1.00 1.00 too 1.00 1.00 1.00 1.00 IN k IN o 1a00 1.00 1,00 1,0 1.00 1.00 1,00 1.00 1.00 1.00 1,00 1.00 1.00 1.00 1,00 1,00 100 1.00 1.00 1.00 1.00 .OD 1 o 1.00 L00 1,00 1.00 Loo 1,00 1.00 1.00 1.00 1.00 1.00 100 1100 1.00 100 1.00 1.00 1.00 I10 1.00 .00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 LODO 90.96 9M8.27 IA.43 102.13 83.61 86.07 80.88 0.00 78.25 75.15 79.53 69.56 0.00 64.31 67.34 69,84 75.22 75 22 78.64 0.00 79.29 80.15 0.00 0.00 76,12 0,00 0.00 59.27 63.51 51.06 0.00 43.97 39.32 41.10 42.25 30.93 0.00 26.81 23.54 27.75 29.52 30.67 29.71 0,00 0.00 26.43 26.69 25.50 0.00 25.53 22,77 22.60 24.47 22.26 25.56 24.42 24.46 25.50 22.74 21.24 25.14 24.30 23.73 22.51 0.00 S-.79-- ATOM 2360 CA ATOM 2361 CB ATOM 2362 001 ATOM 2363 HIt ATOM 2364 C02 ATOM 2365 C ATOM 2366 0 ATOM 2367 N ATOM 2368 H ATOM 2369 CA ATOM 2370 CB ATOM 2371 OGt ATOP- 2372 GI ATOM 2373 C02 ATOM 2374 C ATOM 2375 0 ATOM 2376 N ATOM 237 H ATOM 2378 CA ATOM 2379 CE ATOM 2380 CO ATOM 2381 CD ATOM 2382 OEI ATOM 2383 OE2 ATOM 2384 C ATOM 2385 0 ATOM 2386 39 ATOM 2387 H ATOM 2388 CA ATOM 2389 CB ATOM 2390 CO ATOM 2391 CD ATOM 2392 NH ATOM 2343 HE ATOM 2394 C7, ATOM 2395 N9 ATOM 2396 Hl ATOM 2397 HH12 ATOM 2398 NH2 ATOM 2399 HHZ1 ATOM 2400 HH27 ATOM 2401 C ATOM 2402 0 ATOM 2403 N ATOM 24Q4 H ATOM 2405 CA ATOM 2406 CB ATOM 2407 CGt ATOM 2408 CG2 SO ATOM 2409 C ATOM 2410 0 ATOM 2411 It ATOM 2412 11 ATOM 2413 CA ATOM 2414 CB ATOM 241S OG1 ATOM 2416 H1 ATOM 2417 CGZ ATOM 241$ C ATOM 2419 0 ATOM 2420 N ATOM -2421 H ATOM 2422 CA ATOM 2423 CB

THR

Tma

THR

T4HR

THR

THR

THR

THR

THR

THR

THR

'HR

THR

THR

THR

'HR

GLU

GLU

GLU

GLU

GLU

GL1

GLU

GLU

01-U

GLU

ARG

ARC

ARG

ARG

ARG

ARG

ARC

ARO

ARG

ARG

ARG

ARC

ARG

ARO

ARG

ARO

VAL

VAL

vA.

VAL

VAL

VAL

VAL

VAL

THR

THR

THR

maR

THR

'HR

THR

THR

THR

LE

LEU

LEU

LEU

388 49,646 8.4-18 388 49,037 57.083 388 -48.30i 57.23b 388 49,021 57.669 388 50.105 56.009 388 50.062 58.416 388 49.262 58.508 389 51.357 58.279 389 51,974 58.283 389 51.933 58.099 389 531304 58.762 389 53,043 60.143 389 52.584 60.204 389 54.305 58.266 389 51.981 56.611 389 52.421 55,845 390 51.532 56,183 390 51.265 56.821 390 51,461 54,770 390 49.993 14.334 390 49,716 54.258 390 48,453 53.567 390 47,644 53.290 390 48.274 53.314 390 52,032 54,416 390 52.148 55.236 391 52,406 53,145 391 52.314 52.633 391 52.871 52472 391 51.677 52.087 391 51.351 50.594 391 51,041 50062 391 49,890 49,159 391 49.248 49.211 391 49,642 48.255 391 50.302 48,156 391 51.042 48,794 391 50,046 47,45.7 391 48749 47.269 391 48.279 47.209 391 48.561 46597 391 53842 53.303 391 53.590 53.655 392 54.998 53634 392 55.287 53,159 392 55.880 54,629 392 3&630 55,422 392 57,408 56.628 392 55-579 55918 392 56.865 53.948 392 57.628 53.121 393 56.890 54,255 393 56.183 54.810 393 57.958 53.731 393 57,450 52.681 393 56.238 53,135 393 56,027 52,485 393 57.220 5U33 393 58.633 54.850 393 59.201 54.661 394 59,605 56.072 394 58.080 56.246 394 39.379 57.138 394 59.097 38.483 4.100 5.359 5.956 4.257 2.233 1.316 2.016 2.77t 0709 0,775 0.521 .0321 -0.223 0.446 1.307 -0,742 -1.440 -1,017 -0.902 0.612 1.001 0,129 2,192 -2.35' -3244 -2,449 -1.634 -3.655 548 -4,650 -6,066 -,094 -5.356 -7m066 -8.246 -8.455 4.910 -6.849 -5.968 -7.567 -4,477 -5.630 -3.892 -3,085 -4,494 -3.404 -3.936 -2.436 -5.392 -4915 6,691 -7,081 -7.506 -8.535 -9.148 -9-822 -7,855 -8.235 -9.303 -7.719 -6.913 -8.338 -7.647 23.26 26.35 34.81 0.00 24.27 19.72 20.10 20.14 0.00 22.14 22.05 32.86 0.00 26.26 27.05 23.21 23.67 0.00 21.51 27.17 35.02 38.42 50.83 47.46 22.05 25,04 20,72 0.00 20.99 24.04 27.32 27.94 36.16 0.00 38.21 39.70 0.00 0.00 47.46 0.00 0.00 24.89 24,46 25.48 0.00 21.84 19.22 11.40 18.72 24.62 20.00 23.61 0.00 20.89 21.30 22.68 0.00 17.80 22.52 22.98 19.99 0.00 2.0.6 18.77 t~s:

UM

s0 ATOM 2424 CG ATOM 2425 CDI ATOM 2426 CD? ATOM 2427 C ATOM 2428 0 ATOM 2429 N ATOM 2430 H ATOM 2431 CA ATOM 2432 CB ATOM 2433 OG1 ATOM 2434 501 ATOM 2435 C02 ATOM 2436 C ATOM 2437 0 ATOM 2438 N ATOM 2439 H ATOM 2440 CA ATOM 241 CB ATOM 2442 CG ATOM 2443 Cl) ATOM 2444 NE ATOM 2445 HE ATOM 2446 CZ ATOM 247 N1 25 ATOM 2448 1111 ATOM 2449 1H12 ATOMt 2450 NH2 ATOM 2451 PI21 ATOM 2452 HH122 ATOM 2453 C ATOM 2454 0 ATOM 2455 N ATOM 2456 H ATOM 2457 CA ATOM 245s cB ATOM 2459 SG ATOM 2460 C ATOM2461 0 ATOM 2462 N 40 ATOM 2463 H ATOM 2464 CA ATOM 2465 CB ATOM 2466 CG ATOM 2467 CDI ATOM 2468 CD2 ATOM 2469 CEI ATOM 2470 CE2 ATOM 2471 CS ATOM 2472 C ATOM 2473 0 ATOM 2474 N ATOM 2475 H ATOM 2476 CA ATOM 2477 CR ATOM 2478 CG ATOM 2479 CDI ATOM 2480 CEI ATOM 2431 CDZ ATOM 2482 CEZ EO ATOM 2483 CZ ATOM Z484 OH ATOM 2485 HU ATOM 2486 C ATOM 2487 0

LEV

LEV

LEV

LEV

LEV

THR

TUR

WHR

THR

THR

THR

ARO

THR

ARO

ARO

ARO

ARG

ARO

ARG

ARG

ARG

ARG

ARG

ARO

ARG

ARG

ARG

ARG

AC

ARG

ARG

CYS

CYS

CYS

CYS

CTS

CYS

PRE.

PHE

PRE

PRE

PHE

PHE

PHE

PHE

NEl

PHE

P1R

TYR

TYR

TYR

TYR

TYR

TYR

TYR

TYR

TYR

TYR

TYR

TYR

TYR5 TYR5 394 57.683 59.073 394 57.62 0.395 394 57.320 59.347 394 60.851 56.789 394 61.273 56.518 395 61.659 56.769 395 61.325 56.899 395 63,090 56.503 395 63.474 55.531 395 63.062 56.260 395 63.298 55.742 395 62.748 54.238 395 63.957 57T78 395 65,167 51.754 396 63.362 58.947 396 62.385 58.998 396 64.097 60.187 396 64.189 60.695 396 64.986 59-799 396 64.666 60.158 396 65-266 59.145 396 64.830 58.271 396 66.403 59.385 396 66.982 58.33.

396 66.539 57.441 396 67.836 58.466 396 66.961 60.635 396 66.516 61.394 396 67.815 60.786 396 63-286 61.210 396 62.133 60.990 397 63.80 62.377 397 64766 62545 397 63114 63.404 397 64-086 64447 397 65.267 63.936 397 62170 64.045 397 62.531 64.263 398 60,945 64.386 398 60.638 64.172 398 60072 65.094 398 58,677 6K.374 398 57.644 65.009 398 58.011 65.812 398 56.306 64.770 398 57.052 66.367 398 55.358 65.320 393 55.716 66.121 39S 59.960 66.512 398 59.259 66.853 399 60.703 67.377 399 61.396 67.062 399 60.518 68.809 399 61.878 69.467 399 62.706 69.308 399 63.54 68.55 399 64.366 68.16$ 399 62.601 70.258 399 63.376 70. 173 399 64.262 69.125 399 65.009 68.999 399 65.683 68.322 399 59.536 69.29 399 59.668 69,103 ,78 -7.031 -9.243 -8.181 -7.073 -9.243 -10-160 -9.136 -10.303 11-471 -11'244 -10.343 -9.146 .3,943 -9.392 -9,423 -9.519 -10936 -11,825 -13-248 .1407 -14,151 -14,715 -15.382 -15369 -14722 ,14.247 -15.219 -8.805 -8.483 -8546 830 .7.849 -7.306 -6.027 4.834 -9.977 -8.64 .7.560 -9.380 -9,454 4L0.409 k1494 .10o186 -12.329 -11.028 -12.095 -8.872 926 -9.537 .10.144 -9.395 -9.624 -4.397 -8.293 -7.155 -7.395 -6.258 -6156 -5.005 -5.130 -10.432 -11.630 1.00 1.00 I.Ck 1.00 1.00 1.00 1.00 1,00 1.00 LooO 1.00 1.00 1.00 1.00 1.00 1.00 Looo 1.00 1.00 1.00 1.00 1.00 1.00 IAVo 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 .00 .00 1.00

LODO

1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 100 1.00 1.00 1.00 1-00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 17.38 16.87 17.98 21.53 21.60 24.17 0.00 22.29 19.28 22.57 0.00 15.70 23.93 25.97 23.31 0.00 26.02 28.35 32.64 43.18 51.29 0.00 57.25 62.75 0.00 0.00 55.06 0.00 0.00 25.66 25.00 27.52 0.00 25.75 33.69 39.19 24.55 23.49 21,44 0.00 20.44 22.28 21.70 26.69 18.84 27.Q9 24.85 25.47 22,33 22.03 24.80 0.00 25.99 25.00 27.28 25.30 33.32 30.60 31.42 31.10 35.39 0.00 27.64 23.69 c-n- 81 ATOM 2488 N ATOM 2489 H ATOM 2490 CA ATOM 2491 CB ATOM 2492 CC ATOM 2493 CDI ATOM 2494 CD2 ATOM 2495 C ATOM 2496 O0 ATOM 2497 N ATOM 2498 H ATOM 2499 CA ATOM 2500 CB ATOM 2501 CO ATOM 2502 CDI ATOM 2503 CD2 ATOM 2504 CE1 ATOM 2505 CE2 ATOM 2506 CZ ATOM 2507 C ATOM 2508 0 ATOM 2509 N ATOM 25to10 CD ATOM 2511 CA ATOM 2512 CB ATOM 2513 CO ATOM 2514 C ATOM 2513 0 ATOM 2516 N ATOM 2517 H ATOM 2518 CA ATOM 2519 C ATOM 2520 O0 ATOM 2521 N ATOM 2522 H ATOM 2523 CA ATOM 2524 CB ATOM 2525 CG ATOM 2526 CD2 ATOM 2527 NDIl ATOM 252 HDiD ATOM 2529 CEI ATOM 2530 NEZ ATOM 2531 HE) ATOM 2532 C ATOM 2533 O0 ATOM 2534 OT ATOM 2535 OH2 ATOM 2536 HI, ATOM 2537 R2 ATOM 2538 OH2 ATOM 2539 Ht ATOM 2540 H2 ATOM 2541 OH2 ATOM 2542 H1 ATOM 2543 H2 ATOM 2544 OH2 ATOM 2545 HI ATOM 2546 H2 ATOMN 2547 O2 ATOM 2548 u1 ATOM 2549 112 ATOM 2550 O012 ATOM 2551 Ht

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

LEU

PHE

PHE

PHE

PHE

PHE

PHE

PHE

PHE

FHE

PHE

PHE

PHE

PRO

PRO

PRO

PRO

PRO

PRO

PRO

GLY

GLY

GLY

GLY

GLY

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

HIS

WA7

WAT

WAT

WAT

WAT

WAT

WAT

WAT

WAT

WAT

WAT

WA

WAT

WAT

WAT

WAT

WAT

58,469 69.931 -10.000 58.337 70.072 -9.041 57.598 70.629 -10.934 56.310 71.032 -10.186 55.445 69.821 .9.32 54.671 70.119 -3.568 54.550 69.475 -11.022 5,422 71.858 -11.293 59.402 72.088 -10.612 58.287 72.708 .12.295 57.660 72.565 -13.041 59.210 73.839 -12.269 60.245 73.706 -13.407 61.399 72.857 -12.928 61.474 71.519 -13.260 62.416 73,423 -12,173 62.582 70,764 -12.92 63.512 72.653 -11390 63.611 71.326 -12.167 58.367 75.031 -12.475 58.334 75.584 -13.558 57.638 75.512 -11.527 57751 75.159 -10.125 56.710 76.609 -I11.75 56.137 76.946 -10.380 57161 76.376 -94A28 57.461 7757 -12.427 58.588 78,123 -12.070 56.770 78.302 -13,431 55.816 78.096 -13.558 57.339 79.300 -14.327 58.024 78.691 -15.553 58.363 79.436 -16.458 58.258 77.372 -15.646 57.856 76,702 -15.043 59.038 76.818 -16.735 60.391 76.313 -16.275 61.057 77.377 -15.470 62.077 78,167 -15.915 60.694 77398 -14.262 59.973 77.436 .13.710 61.438 78.828 -13.970 62.266 79.045 -14.963 62.923 79,771 -14,979 58.318 75.633 -11.302 58.804 75.085 -13.288 57.304 73.238 -16.7t1 57,131 53,937 -16. 157 57,956 S3.989 -16.611 56,15 33,157 -16559 59.288 45.222 -12720 59.678 44.463 -12.289 59.326 45.020 -13.638 61.365 66.988 .12.454 61.282 66.754 -11.566 61.920 66.336 -12.878 54.401 52.311 -15.488 53.455 52.320 -15.423 54.685 52.959 -14.831 52.948 645.165 .10.749 53.471 44.927 -9.991 52.622 46.039 -10.552 39.932 72.422 .0.681 40.131 72.039 0.168 24.40 0.00 31.95 27.67 25.95 26.52 24.92 36.21 44.83 36.07 0.00 33.57 32.22 33.80 32.31 39.98 33.77 40.60 36.46 32.19 29.45 29.8 27.11 31.83 27.59 24.71 37.62 38.17 37.83 0.00 40.55 39.74 40.77 40.90 0.00 41.51 43.86 4748 46.94 50.29 0.00 47.33 49.11 0.00 41.65 44.03 41.54 0.00 0.00 24.45 0.00 0.00 18.38 0.00 0.00 26.12 0.00 0.00 22.53 0.00 0.00 41.66 000 g*nsnar*wras~;~ars r~ lll-nra4-uuuar~--~ 82 ATOM 2552 R2 WA? ATOM_2553 0112 WA? ATOW2.554 HI WAT ATOM 2555 l2 WA? ATOM 2556 0HZ WAr ATOM 2557 H1 WAT ATOM 2558 H2 WAT ATOM 2559 0HZ WA? ATOM 2560 Hi WA? .0 ATOM 2561 H2 WA? ATOM 2562 OH2 WAT ATOM Z563 HI WA? ATOM, 2564 HZ WA? ATOM 2565 OH2 WA? 1* .5 ATOM 2566 HI WA? ATOM 2567 R2. WA? ATOM 2568 OHZ WA? ATOM 2569 HI WA? ATOM 2570 HZ WA? ATOM 2571 0112 WA? ATOM 2572 HI WA? ATOM 2573 P12 WA ATOM 2574 OHL WA? ATOM 2575 H1 WAT ATOM 2576 H2 WA? ATOM 2577 012 WA? ATOM 2578 Hi WA? ATOM 2579 HZ WA? ATOM 2580 02 WA? ATOM 2581 H. WAT ATOM 2582 HZ WA? ATOM 2583 OIH' WA? ATOM, 2584 HI WA? ATOM 2585 12 WAT ATOMt 2586 OH2 WA? ATOM 2587 HI WAT ATOM 2588 Hl WA? ATOM 2589 0*1 WAT ATOM 2590 Hi WA? ATOM 2591 H2 WA? ATOM 2592 OH2 WA? ATOM 2593 Hi WA? ATOM 2594 R2 WA? ATOM 259$ 0HZ WA? ATOM 2596. HI WA? ATOM 2597 H? WA? ATOM 2598 OH2 WA? ATOM 2599 1 WA? ATOM 26W RZ WA? ATOM 2601 0H12 WA? ATOM 2602 11 WA? ATOM 2603 HZ WAT ATOM 2604 OHL WA? ATOM 2605 Hi WA? 53 ATOM 2606 HZ WA? ATOM 2607 0HZ WA? ATOM 2608 HI WA? ATOM 2609 E2 WA? ATOM 2610 01ow WA? ,k ATOM 261t Ht WA? ATOM 2612 HZ WA? ATOM 2613 OW2 WAT ATOM 2614 Hi WAT ATOM 2 615 2 WA 261 39,14 262 40.595 262 40.213 262 39.866 263 59.703 263 59.734 263 59.203 264 57.975 264 57.886 264 58.580 265 49.889 265 49.381 265 49.717 266 55.224 266 56.050 266 55.324 267 57.220 267 57.189 267 56.964 268 35.858 26S 36.152 268 35.860 269 48.789 269 47.897 269 49.355 270 59.440 270 59.959 270 58.513 271 48-923 271 48.905 271 49.386 272 44.435 272 44.989 272 44390 273 53.920 273 54.603 273 54232 174 62.871 274 62.632 274 63,467 275 46.942.

275 47,058 275 47.406 276 50,771 276 50.872 276 50.541 277 45.555 277 46.291 272 44.772 278 .066 278 56.509 278 56.806 279 50.499 279 50.711 279 51.009 280 42.398 280 41.614 280 42.691 281 47.580 281 47.146 281 48.027 282 40.354 282 39.834 282 41.237 71.954 65620 65.976 65.614 63.723 63.118 63.239 70.486 69.546 70.685 74.051 73.658 74.986 73.467 73.56W 73.889 72.666 73.-55 71.837 66.670 66.629 67.587 71.28t 71.644 71928 63.444 62.711 63193 44.941 44.016 44.932 59.093 59.185 58.678 52.043 52.710 51.362 68.183 67.292 68,466 70.044 69,784 69-414 63.408 64.350 63.304 65,749 65.423 65.498 46.783 46.154 47.667 65.012 64.&60 65-791 64.363 64.685 65.121 50.658 51.280 50.065 8.806 59.191 58.816 -1.011 4.462 5.270 3.842 -5.839 -5.108 6.512 -7.257 7.232 -6.537 7.407 6.713 7.331 -12,629 -12.183 -13,488 -15.238 -16.021 -15.606 -2.607 -1.699 -2.844 -21,710 -21.837 -22.109 -17.067 -16.814 -17.097 -15.001 -14,762 -15.826 9.639 8.817 10.3W -8,038 4Q-021 -4.608 -1.698 -1.909 -2.393 -23.874 -24,775 -23.326 17.889 17.94 16.967 12.972 12.436 12518 1 1 -Z.204 -12.005 -29411 -23.494 -29.571 -27.639 -28.072 -27. 14$ -26.241 -26.06 -26-836 -25.242 -25.588 1.00 1.00 .0o 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 I.00 1.00 1.00 1.00 1.00 Louo 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 .00 '100

.OD

1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 Z.00 1.00 1.00 1.00 1.00 t100 1.00 100 1.00 1.00 1.00 1-00 1.00 t.00 1.00 1.00 1.00 1.00 1.00 0.00 27 37 0.00 0.00 23.97 0.00 0.00 26.26 0.00 0.00 0.00 0.00 54,87 0.00 0.00 33.22 0.00 0.00 29.58 0.00 D.00 49.48 0.00 0.00 23.58 0.00 0.00 59.67 0.00 0.00 24.08 0.00 0.00 0.00 0.00 38.92 0.00 0.00 49.11 0-00 0.00 54.39 0-00 0.00 0.00 0.iK0 27.22 0-00 0.00 47.42 0.00 73-43 0.00 0-00 45.60 0.00 0.00 51.46 0.00 0.00 83 ATOM 2616 0182 WA? 283 59.582 57.507 -22.297 1.00 50.06 ATOM 2617 iL WA? 283 59.205 58.060 -22.973 1.00 000 ATOM 2619 H2 wAT 283 6.6 ATOM 215 82 WAT? 283 60 461 57.302 -22574 1.00 000 ATOM 2619 HZ WA? 284 62787 66.431 -21.929 1.00 64.67 ATOM 2620 H1 WAT 284 62.786 67.378 -21.776 1.00 0.00 ATOM 2621 H2 WAT 254 63.310 66.298 -22.709 1.00 0.00 ATOM 2622 82 WAT 285 42.178 68.676 -21.635 .00o 47.92 ATOM 2623 HI WA? 285 42.226 68.152 -20.834 1.00 0.00 ATOM 2624 2 WA 285 41.525 68.220 -22.154 1.00 000 ATOM 2625 OH2 WA? 286 59.860 64.459 -20.626 1 o 45.90 ATOM 2626 HI WAT 286 59.529 63.613 -20.882 1.00 0 00 ATOM 2627 H2 WA? 286 60.034 64.935 -21.429 1.00 0-00 ATOM 2628 8 WAT 287 3&5SC 60.429 -19.380 1.00 45.23 ATOM 2629 1 WAT 287 37.765 59.995 -19.240 1-00 0.00 S 15 ATOM 2630 H2 WAT 287 38.339 61.266 -19.751 1.00 000 ATOM 2631 OH2 WAT 288 49.737 64.079 -19.712 1.00 31.09 ATOM 2632 81 WA? 288 49.889 64.551 -20.540 1.00 0.00 ATOM 2633 H2 WAT 288 48.791 63.991 -19.646 1.00 0.00 ATOM 2634 O82. WAT 29 45.077 74.284 -19.985 1.00 5831 ATOM 2635 111 WAT 289 44.803 75.186 -19.856 100 000 ATOM 2636 HZ WAT 289 45.225 73.984 -19.086 1.00 0.00 ATOM 2637 2 WA 290 36.463 6896 -18.372 1.00 41.38 ATOM 2638 HI WA 290 36.206 68.878 -19.244 1.00 000 ATOM 2639 H2 WAT 290 3730 69.021 -18.240 1.00 0-00 25 ATOM 2640 OH2 WAT 291 42.%09 7.175 -18360 1.00 300 ATOM 2641 HI WAT 291 42.18 73.868 -17.902 1.00 000 ATOM 2642 2 WAT 291 41.926 72.928 -19.071 1.00 0.00 ATOM 2643 12 WAT 292 52.772 54.057 -17.864 1.00 34.00 ATOM 2644 HI wAT 292 52.675 54.050 -16.919 1.00 0.00 ATOM 264 H2 WAT 292 53.709 5413 -17.997 1.00 0.00 ATOM 2646 OH2 WA? 293 58.499 51.54 -17.264 1.00 39.19 SATOM 2643 13 WA? 293 58.96 12 17.042 1.00 000 ATOM 264 H2 WAT 293 59.231 52A21 -17.413 1.00 0.00 35 ATOM 2649 012 WA 294 55.293 76.832 -15.365 1.00 n96 3 ATOM 2650 HI WAT 294 55.84 76689 -14489 1.00 00 ATOM 2651 O2 WA? 294 55.894 76.271 -15.896 1.00 000 ATOM 2652 01l WAT 295 50254 47.990 -11.950 1.00 3408 ATOM 2653 HE WAT 295 49,709 48.063 -12.721 1.00 000 ATOM 2634 HI WA? 295 49,755 47.316 -11.486 1.00 0.00 40 ATOM 2655 OH2 WAT 296 37.749 48.038 1.00 50.16 ATOM 2656 111H WA? 296 36.805 48.072 4-973 1.00 000 ATOM 2657 H2 WAT 296 37.815 47.302 -9.501 1.00 0.00 ATOM 2658 OH2 WAT 297 615 72.978 -8.83 1.00 30.80 SATOM 2659 H1 WAT 297 62.021 72.636 821 1.00 0.00 ATOM 2660 12 WA 297 61.120 73.515 -8.3 1.00 0.00 ATOM 2661 OH2 WA? 298 46.716 77.808 -7.02 1.00 36 58 ATOM 2662 HI WA? 295 47.000 78.075 -6.217 1.00 0.00 ATOM 2463 H2 WAT 298 47.380 78.176 -7.649 1.00 0-00 ATOM 2664 OH2 wAT 299 43.918 76.808 01 1.00 35-53 ATOM 2665 1 WAT 299 43.8 77.750 4-6.052 1.00 0.00 ATOM 2666 H2 WAT 299 44.809 76.650 782 1.00 000 ATOM 2667 0oH2 WAT 300 60.882 61.010 -5.831 1.00 3221 ATOM 2668 1I WAT 300 60547 60.543 -5.m 1.00 0.00 ATOM 2669 H2 WAT 300 60943 61.933 -5.683 1.00 000 ATOM 2670 OH2 WAT 301 56.234 77.47 -5.323 1.00 4588 ATOM 2671 1 WAT 301 55.449 77.454 4.859 1.00 000 ATOM 2672 H2 WAT 301 56.348 77.543 -5.971 1.00 000 ATOM 2673 012 WAT 302 43.603 47.976 -4.116 1.00O 46.63 ATOM 2674 HI WAT 302 43.969 48.651 -4654 1.00 0.00 ATOM 2675 HZ WAT 302 43.745 47.160 601 I.0 0.00 ATOM 2676 012 WAT 303 41.t12 55.660 -0.536 1.00 36.50 ATOM 2677 Rt wAT 303 41.333 54.851 -0.1 10 0.00 ATOM 2678 H2 WAT 303 42.325 55.359 -1193 100 0.00 ATOM 2679 O2 WA? 304 51.729 51.!56 -0590 100 72.02 -I c R--IIl--~T*e L-ru, 84 &TOM 2680 HI WAX ATOM 2681 12 WAT ATOM 2682 OH2 WAX ATOM 2683 HI WAX S ATOM 2684 H2. WAX ATOM 2635 OH2 WAT ATOM 2686 HI WAX ATOM 2687 H2 WAX ATOM 2688 0H2 WAT ATOM 2689 Hl WAX ATOM 2690 HZ WAr ATOM 2691 OHZ WAT ATOM 2692 HI WAX ATOM 2693 HZ WAX ATOM 2694 0HZ WAX li, ATOM 2695 Hi WAX ATOM 2696 2 WAT ATOM 2697 OH2 WAT ATOM 268 RI WAT ATOM 2699 HZ wAr ATOM 2700 OHZ WAX ATOM 201 HI WA ATOM 2702 H2 WAX ATOM 2703 0HZ WAT ATOM 2704 H1 WAX ATOM 205 H WAX ATOM 2706 0HZ. WAX ATOM 2707 Hl WAT ATOM 2708 112 WAX ATOM 2709 0HZ WAX ATOM 2710 HI WAX ATOM 2711 HZ WAX ATOM 2712 OHZ WAX ATOM 2713 HI WAX ATOM 2714 H WAX ATOM 715 OH2 WAX ATOM 2716 HI WAX ATOM 2717 HZ WAX ATOM 271 OH12 WAX ATOM 719 Hi WAX ATOM 2720 HZ. WAX ATOM 272 O2 WAX ATOM 2722 HI WAX ATOM 2723 H2 WAX ATOM 24 oH WAX ATOM 275 H! WAX ATOM 2726 H2 WAT ATOM 2727 012 WAr ATOM 2728 Hl WA ATOM 2729 H2 WAX ATOM 2730 oH2 WA ATOM 2731 Hl WAX ATOM 272 HZ WAX ATOM 2733 OH2 WAT ATOM 2734 HI WAX' ATOM 2735 HZ WAX ATOM 2736 OH WAX ATOM 2717 Ht WAX ATOM 2736 Hl WAX AxoM z739 01 WAT ATOM 2740 HI WAX ATOM 2741 HZ WAX ATOM 2742 0H2 WAX ATOMV- 2743 HI WAX 304 52.459 50.567 304 51.363 51.290 305 4.576 76.180 305 44.696 75.258 305 44.178 76.553 306 36.913 54.669 306 39.203 55.452 306 38.207 54.284 307 42.134 58.338 307 41.312 57.982 307 42.564 57.551 308 56.646 60.941 308 55.700 0.977 306 56.929 61.839 309 45.030 48.554 309 44.943 47.651 309 45.909 48.606 310 41.590 59.650 310 41.965 59.981 310 41.171 60.430 311 30.67$ 62.812 311 31.519 63.059 311 30.787 61.904 312 44.035 51.425 312 43,759 1.313 312 43.889 50.557 313 53.084 69.483 313 53.66 68.732 313 52.585 69.526 314 33.280 54.578 314 32.487 54.073 314 33.372- 54.689 315 60.509 60,787 315 60.849 61.538 31 60.079 61,112.

316 36.436 51.291 316 36.114 50,786 316 35.650 5162 317 47.54W 66.402 317 46.808 66.985 317 48149 66.611 318 39.908 61.653 318 39.811 62.356 31& 40.435 61.009 319 43.648 51.317 319 44.217 51.632 319 42.796 51.245 320 42.904 66.185 320 43.844 66.182 320 42.797 66.244 321 52.576 73.792 321 52.246 73.438 321 51.924 73.486 322 61.556 50.185 322 60.747 49.697 322 62.307 49.596 323 24.851 56.075 323 25.114 55.419 323 25.365 55.812 324 30.036 71.409 324 29.363 70.770 324 29.785 71.599 325 33.127 79.276 325 33.528 79.402 -0.423 0.284 0.070 -0-493 0.203 -0.215 .0306 1,150 1.511 0.838 0.737 0.666 0.583 91.92 9.474 8.834 10.886 10.534 10.599 10.280 10.876 12.63 13.40 13,409 12,474 14.147 14.271 13.208 -18.332 -18.810 -17.565 10.254 9.515 10,727 -29.189 -29.300 -29.900 -21.569 -20.946 -21.097 20053 -19-345 -19.643 -19.404 -19.470 .18.46 -19.312 -18.497 -19.932 46-806 -16.824 -16.932 -3.153 -3.916 -2744 -3-022 -1.852 0.558 -0.294 0.00 0.00 70.30 0.00 0.00 39.19 0.00 0.00 29.79 0.00 0.00 3S.97 0.00 0.00 48,96 0.00 0.00 32.65 0.00, 0.00 48.30 0.00 49.62 45.11 0.00 0.00 30.29 0.00 30.00 49.82 0.00 0.00 49.95 0.00 0.00 55.32 0.00 0.00 55.22 0.00 0.00 43.44 0.00 0.00 47.B8 0.00 0.00 W0.71 0.00 0.00G 39.05 0.00 1100 45.31 0.00 0.00 0.00 0-00 O-W4 57-23 Q-W0 .TOM 2744 2 WAT 325 32.60 80.047 0.708 1.00 0.00 ATOM 2745 011OH2 WAT 326 35.907 47.459 2.721 1.00 52.14 ATOM 2746 H1 WAT 326 35.224 43.120 2.665 100 0.00 ATOM 2747 82 WAT 326 36.753 47.864 2.620 1.00 0.00 ATOM 2748 OH WAT 327 54.215 72.016 6.546 t.00 52.02 ATOMt 2749 HI WAT 327 55.027 71.530 6.405 1.00 0.00 ATOM 2750 H2 WAT 327 54.516 72.859 6.883 1.00 0.00 ATOM 2751 OH1 WAT 328 41.269 52.487 1.122 1.00 51.87 ATOM 2752 HL WAT 328 40.694 51.781 1.440 1.00 0.00 ATOM 7753 H2 WAT 328 42.127 52.127 1.259 100 0.00 ATOM 2754 012. WAT 329 34.066 58.806 13.164 100 5571 ATOM 2755 R1 WAT 329 34.724 59.474 13.292 1.00 0.00 ATOM 2756 H2 WAT 329 34.564 58.010 12.945 1.00 0.00 ATOM 2757 0H WAT 330 41.316 52.756 13.918 1.00 44.03 ATOM 2758 HRI WAT 330 42.281 53.408 13.395 100 Q 0.00 ATOM 2759 812 WAT 330 42.525 52.335 14.395 1.00 0.00 ATOM 2760 OH2 WAT 331 39.370 62.098 14.302 1.00 54.70 ATOM 2761 RI WAT 331 38.736 62.661 14.727 1.00 0.00 ATOM 2762 H2 WAT 331 39.761 62.569 13.574 !.00 0.00 ATOM 1763 OH WAT 332 50.309 69.365 13364 1 00 54.23 ATOM 2764 H3 WA 332 50.055 69.719 12.508 00 0.00 ATOM 2765 H1 WAT 332 51.043 69.910 13.645 1.00 0-00 ATOM 2766 02H WAT 333 40.562 55.451 15.773 1.00 61.39 ATOM 2767 RI WAT 333 39723 55.080 16.041 1.00 0.00 ATOM 2768 H2 WAT 333 40.748 55.017 14.937 1.00 0.00 The following abbreviations are used in Table B.

"Atom type" refers to the element whose coordinates are measured. The first letter in the column defines the element.

Y, Z" crystallographically define the atomic position of the element measured "Bt is a thermal factor that measures movement of the atom around its atomic center.

Atoms numbered 153-158 CLys-14) and 184-189 35 (Ser-149) were modeled as Ala residues.

Atoms numbered 1487-1534 and designated "Ald" in the column titled "Residue" are Cys-285 bound to the tetrapeptide aldehyde inhibitor.

Structure cocirdinates for ICE according to Table B may be modified from this original set by mathematical manipulation. Such manipulations include, but are not limited to, crystallographic permutations of the raw structure coordinates, fractionalization of the raw structure coordinates, integer additions or subtractions to sets of the raw structure coordinates, 86inversion of the raw structure coordinates, and any combination of the. above.

-86 inversi~on of the raw structure coordinates, and any combination of the above, I_ _U 87 SEQUENCE LISTING GENERAL INFORMATION:

APPLICANT:

NAMnE: Vertex Pharmaceuticals, Inc.

STREET: 40 Allston Street CITY: Cambridge STATE: Massachusetts COUNTRY: United States of America POSTAL CODE (ZIP): 02139 TELEPHONE: 517-576-3111 TELEFAX: 617-576-2109 (ii) TITLE OF INVENTION: CRYSTAL STRUCTURE AND MUTANTS OF INTERLEUKIN-1 BETA CONVERTING ENZYME S (iii) NUMBER OF SEQUENCES: 1 (iv) COMPUTER READABLE FORM: MEDIUM TYPE: Ploppy disk COMPUTER: IBM PC compatible OPERATING SYSTEM: PC-DOS/MS-DOS SOFTWARE: PatentIn Release Version #1.30 (EPO) (vi) PRIOR APPLICATION DATA: APPLICATION NUMBER: US 08/261.582 FILING DATE: I7-JUN-1994 INFORMATION FOR SEQ ID NO: 1: SEQUENCE CHARACTERISTICS: LENGTH: 404 amino acids TYPE: amino acid K STRANDEDNESS: TOPOLOGY: linear (ii) MOLECULE TYPE: protein (iii) HYPOTHETICAL: NO (xi) SEQUENCE DESCRIPTION; SEQ IW NO: 1: Met Ala Asp Lys Val Leu Lys Glu Lys Arg Lys Leu Phe le Arg Ser 1- 5 10 1 so i Met Gly Glu Gly 7hr Ile Asn Gly Leu Leu Asp Glu Leu Leu Gln Thr 25

II

I--m Arg Val Leu Asn Lys Glu Glu Thr Val Met Asp Lys Thr Arg o 4 2 5 4.

4 Gly Ala Ser Tyr Asa Tyr Ala Pro Ser Glu 130 Trp LyS 145 Arg Thr Pro Arg Leu Gin Set Asp 210 Lys Thr 22$ Glu Gly Gin Leu Leu Lys Gin Ala Cys Gin Ile 70 Leu Ala Gly Th' Len Leu Asn Met Gin Asp 100 Gln Ala Val Gin Asp 115 Gly Asn Val Lys Len 135 Gin Lys Ser Ala Glu 150 Arg Leu Ala Leu Ile 165 Arg Thr Gly Ala Glu 180 Asn Len Gly Tyr Ser 195 Met Thr Thr Glu Leu 215 Ser Asp Ser Thr Phe 230 Ile Cys Gly Lys Lys 245 Asn Ala Ile Phe Asn 260 Asp Lys Pro Lys Val 275 88 Met Glu Lys Val Lye Axg Gin 40 Ala Leu Lie Asp Ser Val Ile Cys lie Thr Tyr Ile ys Glu 75 Gly Leu Ser Ala Asp Gin Thr Ser Gin Gly Val Leu Ser Set 105 110 Asn Pro Ala Met Pro Thr Sr 120 125 Cys Ser Leu Glu Glu Ala Gin 140 Ile Tyr Pro Ile Met Asp Lye 155 Ile Cys Asn Glu Glu Phe Asp 170 Val Asp Ile Thr Gly Met Thr I 185 190 Val Asp Val Lye Iys Asn Leu 200 205 Glu Ala Phe Ala His Arg Pro 220 Len Val Phe Met Ser His Gly 235 His Ser Glu Gin Val Pro Asp 250 Met Leu Ass Thr Lye Asn Cys 265 270 Ile Ile Ile Gin Ala Cys Arg 280 285 Lys Asp Set Va Giy Val Ser 300 Asn Ala Pro Lys Glu Asp Ser Gly Phe Pro Set Gly Arg Ile Ser Set 160 Ser Ile 175 Met Len Thr Ala Glu His Ile Arg 240 Ile Leu 255 Pro Ser Gly Asp Gly Asn Ser Pro Gly Val Val Trp Phe 290 295 Set Leu Pro Thr Thr Glu Gu Phe Glu Asp Asp Ala Ile Lys Lys 310 315 320 a i f i i i Otis 89 Ala His Ile G3.u Lys Asp phe Ile Ala Phe cys Ser Ser Tkir Pro Asp 325 330 335 Asa Vakl Ser Trp Arg His Pro Thr M4et (31y Ser Val Phe Ile Gly Arg 340 345 350 L~eu Ile Glu His met G-in Glu- Tyr Ala CYS Ser Cys Asp Val Glu Glu 355 360 365 Ile Plie Arg Lys Val Arg Phe Ser Pkie Glu Gin Pro Asp Gly Arg Ala 370 375 380 Gin Met Pr~o Thr Thr Glu Arg Val. Thr Leu Thr Arg Cys Phe Tyr Leu 385 390 395 400 Phe Pro Gly His '66 6 6 6 6 a I 6 £6 6 £4 Ia 4 £6

WI

Claims (9)

1. An interleukin-19 converting enzyme, wherein one or more of the amino acids in the active site or in the accessory binding site are replaced by one or more amino acids selected from the group I0 consisting of naturally occurring amino acids, unnatural amino acids, selenocysteine and

2. The interleukin-If. converting enzyme according to claim 1, wherein a hydrophilic or hydrophobic amino acid residue in said active site or said accessory binding site is replaced. selno th nne 3 The interleukin-lg converting enzyme 20 according to claim 1, wherein said active site amino acid is selected from the group consisting of amino acids 173, 176, 177, 178, 179, 180, 236, 237, 238, 239, 244, 248, 283, 284, 285, 290, 338, 339, 340, 341, 342, 343, 345, 348, 352, 381 and 383 of SEQ ID NO:l. .4*

4. The interleukin-1 converting enzyme according to claim 1, wherein said accessory binding site amino acid is selected from the group consisting of amino acids 150, 151, 240, 259, 267, 268, 274, 291, 292, 293, 294, 295, 296, 297, 317, 318, 319, 320, 321, 322, 323, 324, 325, 327, 334, 335, 367, 371, 374, 375, 377, 378, 380, 382, 384, 386, 388, 389, 390, 391, 392, 393, 394, 395 and 396 of SEQ ID NO:i. a 91 The interleukin-19 converting enzyme according to claim 1, wherein at least one cysteine amino acid is replaced by an amino acid selected from the group consisting of selenocysteine or selenomethionine.

6. The interleukin-1 converting enzyme according to claim 1, wherein at least one methionine amino acid is replaced by an amino acid selected from 10 the group consisting of selenocysteine or selenomethionine.

7. The interleukin-1l converting enzyme according to any one of claims 1 to 6, wherein said 15 enzyme is in crystalline form.

8. The interleukin-lf1 converting enzyme according to claim 1, wherein said enzyme is characterized by increased stability to subunit 20 dissociation.

9. The interleukin-1l converting enzyme according to claim 1, said enzyme having higher specific activity than the wild-type enzyme. The interleukin-lg converting enzyme according to claim 1, said enzyme having altered substrate specificity.

11. The use of an interleukin-l1 converting enzyme according to claim I to determine binding interactions between a chemical compound and the enzyme. -92-

12. An interleuk-ifl-l Conrverting enzyme, wherein at least one aminao acid residue on, at or near the surface of- said enzyme is replaced, resulting in an altered surface charge o-F one or more charge units. DATED this 8th day of December 1998 VERTEX PHARMACEUTICALS INCORPORATED By their Patent Attorneys CULLEN &CO.