CA2346929A1 - A modified polypeptide with reduced immune response - Google Patents

A modified polypeptide with reduced immune response Download PDF

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CA2346929A1
CA2346929A1 CA002346929A CA2346929A CA2346929A1 CA 2346929 A1 CA2346929 A1 CA 2346929A1 CA 002346929 A CA002346929 A CA 002346929A CA 2346929 A CA2346929 A CA 2346929A CA 2346929 A1 CA2346929 A1 CA 2346929A1
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atom
polypeptide
gly
ala
amino acid
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Arne Agerlin Olsen
Claus Von Der Osten
Kim Vilbour Andersen
Steffen Ernst
Erwin Ludo Roggen
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Novozymes AS
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    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A61K47/646—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/37—Polymers
    • C11D3/3703—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
    • C11D3/3719—Polyamides or polyimides
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
    • C12N9/2411—Amylases
    • C12N9/2414—Alpha-amylase (3.2.1.1.)
    • C12N9/2417—Alpha-amylase (3.2.1.1.) from microbiological source
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C12N9/54—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea bacteria being Bacillus

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Abstract

The present invention relates to polypeptides with reduced immune response including reduced allergenicity having one or more amino acid residues being substituted with other amino acid residues and/or having coupled one or more polymeric molecules in the vicinity of the polypeptides metal binding site, a method for preparing modified polypeptides of the invention, the use of said polypeptide for reducing the immunogenicity and allergenicity and compositions comprising said polypeptide.

Description

WO 00/22103 .PCT/DK99/00542 Title: A MODIFIED POLYPEPTIDE WTTH REDUCED IMMUNE RESPONSE
FIEhD OF THE INVENTION
s The present invention relates to polypeptides having substituted one or more amino acid residues to said polypeptide and/or having coupled polymeric molecules on the surface of the 3-dimensional structure of the polypeptide, a method--for preparing modified polypeptides of the invention, the use of io said modified polypeptides for reducing the immunogenicity and allergenicity, and compositions comprising said polypeptide.
BACKGROUND OF THE INVENTION
The use of polypeptides, including enzymes, in the is circulatory system to obtain a particular physiological effect is well-known in the medical arts. Further, within the arts of industrial applications, such as laundry washing, textile bleaching, personal care, contact lens cleaning, and food and feed preparation enzymes are used as a functional ingredient.
2o One of the important differences between pharmaceutical and industrial application is that for the latter type of applications (i.e. industrial applications) the polypeptides (often enzymes) are not intended to enter into the circulatory system of the body.
2s Certain polypeptides and enzymes have an unsatisfactory stability and may under certain circumstances - dependent on the way of challenge - cause an immune response, typically an IgG
and/or IgE response.
It is today generally recognized that the stability of 3o polypeptides is improved and the immune response is reduced when polypeptides, such as enzymes, are coupled to polymeric molecules. It is believed that the reduced immune response is a result of the shielding of (the) epitope(s) on the surface of the polypeptide responsible for the immune response leading to as antibody formation by the coupled polymeric molecules.
Techniques for conjugating polymeric molecules to polypeptides are well-known in the art.

WO 00/22103 .PCT/DK99/00542 One of the first suitable commercially techniques was described back in the early 1970'ies and disclosed in e.g. US
patent no. 4,179,337. Said patent concerns non-immunogenic polypeptides, such as enzymes and peptide hormones coupled to s polyethylene glycol (PEG) or polypropylene glycol (PPG). At least 15% of the polypeptides' physiological activity is maintained.
GB patent no. 1,183,257 (Crook et al.) describes chemis-try for conjugation of enzymes to polysaccharides via a triazine io ring.
Further, techniques for maintaining of the enzymatic activity of enzyme-polymer conjugates are also known in the art.
WO 93/15189 (Veronese et al.) concerns a method for maintaining the activity in polyethylene glycol-modified is proteolytic enzymes by linking the proteolytic enzyme to a macromolecularized inhibitor. The conjugates are intended for medical applications.
It has been found that the attachment of polymeric molecules to a polypeptide often has the effect of reducing the activity ao of the polypeptide by interfering with the interaction between the polypeptide and its substrate. EP 183 503 (Beecham Group PLC) discloses a development of the above concept by providing conjugates comprising pharmaceutically useful proteins linked to at least one water-soluble polymer by means of a reversible 2s linking group.
EP 471,125 (Kanebo) discloses skin care products comprising a parent protease (Bacillus protease with the trade name Esperase~) coupled to polysaccharides through a triazine ring to improve the thermal and preservation stability. The coupling ao technique used is also described in the above mentioned GB
patent no. 1,183,257 (Crook et al.).
JP 3083908 describes a skin cosmetic material which contains a transglutaminase from guinea pig liver modified with one or more water-soluble substance such as PEG, starch, as cellulose etc. The modification is performed by activating the polymeric molecules and coupling them to the enzyme. The composition is stated to be mild to the skin.
WO 98/35026 (Novo Nordisk A/S) describes polypeptide-polymer conjugates having added and/or removed one or more attachment groups for coupling polymeric molecules on the surface of the polypeptide structure. The conjugates have reduced immunogenicity and allergenicity.
SU'~ARY OF THE INVENTION
It is the object of the present invention to provide improved polypeptides suitable for industrial and pharmaceutical applications.
io The term "improved polypeptides" means in the context of the present invention polypeptides having a reduced immune response in humans and animals. As will be described further below the immune response is dependent on the way of challenge.
The present inventors have found that polypeptides, such as i5 enzymes, may be made less immunogenic and/or allergenic by substituting one or more amino acid residues on the surface of the polypeptide with other amino acid residues and/or by coupling polymeric molecules on the surface of the enzyme in the vicinity of a bound ligand of the enzyme e.g. a metal ion zo substantially without affecting the enzymatic activity.
When introducing pharmaceutical polypeptide directly into the circulatory system (i.e. bloodstream) the potential risk is an immunogenic response in the form of mainly IgG, IgA and/or IgM antibodies. In contrast hereto, industrial polypeptides, i5 such as enzymes used as a functional ingredient in e.g.
detergents, are not intended to enter the circulatory system.
The potential risk in connection with industrial polypeptides is inhalation causing an allergenic response in the form of mainly IgE antibody formation.
ao Therefore, in connection with industrial polypeptides the potential risk is respiratory allergenicity caused by inhala-tion, intratracheal and intranasal presentation of polypeptides.
The main potential risk of pharmaceutical polypeptides is immunogenicity caused by intradermal, intravenous or subcu 35 taneous presentation of the polypeptide.
The term "immunogenicity" used in connection with the present invention may be referred to as allergic contact dermatitis in a clinical setting and is a cell mediated delayed immune response to chemicals that contact and penetrate the skin. This cell mediated reaction is also termed delayed contact hypersensitivity (type IV reaction according to Gell and Combs s classification of immune mechanisms in tissue damage).
The term "allergenicity" or "respiratory allergenicity" is initially an immediate anaphylactic reaction (type I antibody-mediated reaction according to Gell and Combs) following inhalation of e.g. polypeptides.
io According to the present invention it is possible to provide polypeptides with a reduced immune response, which has a substantially retained residual activity.
The allergic and the immunogenic response are in one term, at least in the context of the present invention called the i5 "immune response".
In the first aspect the invention relates to a polypeptide with reduced immune response, having one or more amino acid residues modified, wherein the C"-atoms of said amino acid residues are located less than 15 A from the ligand bound to ao said polypeptide.
The reduced immune response is preferably reduced allergenicity.
The modification of the polypeptide is conducted by substituting one or more amino acid residues in the parent 25 polypeptide with other amino acid residues to said polypeptide, and/or by selecting variants from a diverse library of variants of the parent polypeptide and/or by coupling a polymeric molecule to the surface of the parent polypeptide.
The term "parent polypeptide" refers to the polypeptide to 3o be modified by coupling to polymeric molecules or by substituting amino acid residues. The parent polypeptide may be a naturally-occurring (or wild-type) polypeptide or may be a variant thereof prepared by any suitable means. For instance, the parent polypeptide may be a variant of a naturally-occurring 35 polypeptide which has been modified by substitution, deletion or truncation of one or more amino acid residues or by addition or insertion of one or more amino acid residues to the amino acid sequence of a naturally-occurring polypeptide.

A "suitable attachment group" means in the context of the present invention any amino acid residue group on the surface of the polypeptide capable of coupling to the polymeric molecule in question.
s Preferred attachment groups are amino groups of Lysine residues and the N-terminal amino group. Polymeric molecules may also be coupled to the carboxylic acid groups (-COOH) of amino acid residues in the polypeptide chain located on the surface.
Carboxylic acid attachment groups may be the carboxylic acid 1o group of Aspartate or Glutamate and the C-terminal COON-group.
Another attachment group is SH-groups in Cysteine.
An "active site" means any amino acid residues and/or molecules which are known to be essential for the performance of the polypeptide, such as catalytic activity, e.g. the catalytic is triad residues, Histidine, Aspartate and Serine in Serine proteases, or e.g. the heme group and the distal and proximal Histidines in a peroxidase such as the Arthromyces ramosus peroxidase.
A "ligand", means in the context of the present invention a zo metal or metal ion or a cofactor.
In the context of the present invention "modification of amino acid residues" means that amino acid residues are substituted with other amino acid residues and/or a polymeric molecule is coupled to the amino acid residue. The polypeptide 2s of the present invention may according to the invention be modified by substitution alone, by coupling of a polymeric molecule alone or by a combination of substitution and coupling.
In the context of the present invention "located" means the shortest distance from any atom in the ligand to the relevant C
3o atom in the amino acid residue.
Furthermore, in the context of the present invention e.g.
"R250K" means that the amino acid Arginine in position 250 of the polypeptide has been substituted with the amino acid Lysine according to the one-letter-code of amino acids.
35 in the second aspect the invention relates to a method for preparing polypeptides with reduced immune response comprising the steps of:
a) identifying amino acid residues located on the surface of the 3-dimensional structure of the parent polypeptide in question, b) selecting target amino acid residues on the surface of said 3-dimensional structure of said parent polypeptide to be modified, s c) substituting one or more amino acid residues selected in step b) with other amino acid residue, and/or d) coupling polymeric molecules to the amino acid residues in step b) and/or step c).
The invention also relates to the use of a modified io polypeptide of the invention and the method of the invention fox reducing the immunogenicity of pharmaceuticals and reducing the allergenicity of industrial products.
Finally the invention relates to compositions comprising a modified polypeptide of the invention and further ingredients is used in industrial products or pharmaceuticals.
BRIEF DESCRIPTION OG THE DRAWINGS
Figure 1 shows integrated IgE antibudy levels in rats.
Figure 2 shows integrated specific IgE levels in mice.
DETAILED DESCRIPTION OF THE INVENTION
It is the object of the present invention to provide improved polypeptides suitable for industrial and pharmaceutical applications.
Even though polypeptides used for pharmaceutical applications and industrial application can be quite different the principle of the present invention may be tailored to the specific type of parent polypeptide (i..e. enzyme, hormone peptides etc.).
The present inventors have found that polypeptides, such as ao enzymes, may be made less immunogenic and/or less allergenic by substituting amino acid residues in the vicinity of the ligand e.g. metal ion at the metal ion binding site and/or by coupling one or more polymeric molecules on the surface of the parent polypeptide. In addition thereto the inventors have found that a high percentage of maintained residual catalytic activity may be maintained in these modified polypeptides.
In the first aspect the invention relates to an improved polypeptide having one or more amino acid residues modified, wherein the C°'-atom of said amino acid residues is located less than 15 ~ from the ligand bound to said polypeptide.
The substitution of amino acid residues and coupling of polymeric molecule may be carried out in a conventional manner s as described below.
Reduced immune response vs. maintained residual enzymatic activity For enzymes, there is a conflict between reducing the immune io response and maintaining a substantial residual enzymatic activity.
Without being limited to any theory it is believed that the loss of enzymatic activity of enzyme-polymer conjugates might be a consequence of impeded access of the substrate to the active is site in the form of spatial hindrance of the substrate by especially bulky and/or heavy polymeric molecules to the catalytic cleft. It might also, at least partly, be caused by disadvantageous minor structural changes of the 3-dimensional structure of the enzyme due to the stress made by the coupling zo of the polymeric molecules.
Also, polypeptides modified by substituting one or more amino acid residues may have reduced enzymatic activity.
Maintained residual activity 2s A modified polypeptide of the invention has a substantially maintained catalytic activity.
A "substantially" maintained catalytic activity is in the context of the present invention defined as an activity which is above 20%, at least between 20% and 30%, preferably between 30%
3o and 40%, more preferably between 40% and 60%, better from 60% up ' to 80%, even better from 80% up to about 100%, in comparison to the activity of the modified polypeptide prepared on the basis of corresponding parent polypeptides.
In the case of polypeptide-polymer conjugates of the ss invention where no polymeric molecules are coupled at or close to the active sites) the residual activity may even be up to 100% or very close thereto. If attachment groups) of the parent polypeptide is(are) removed from the active site the activity might even be more than 100 in comparison to modified (i.e.
polymer coupled) parent polypeptide conjugate.
The attachment Group s Virtually all ionized groups, such as the amino groups of Lysine residues, are located on the surface of the polypeptide molecule (see for instance Thomas E. Creighton, (1993), "Proteins", W.H. Freeman and Company, New York).
Therefore, the number of readily accessible attachment groups io (e. g. amino groups) on a modified or parent polypeptide equals generally the number of Lysine residues in the primary structure of the polypeptide plus the N-terminus amino group.
The chemistry of coupling polymeric molecules to amino groups is quite simple and well established in the art.
is Therefore, it is preferred to add Lysine residues (i.e.
attachment groups) to the parent polypeptide in question to obtain improved conjugates with reduced immunogenicity and/or allergenicity and/or improved stability and/or high percentage maintained catalytic activity.
zo Polymeric molecules may also be coupled to the carboxylic groups (-COOH) of amino acid residues on the surface of the polypeptide. Therefore, if using carboxylic groups (including the C-terminal group) as attachment groups addition and/or removal of Aspartate and Glutamate residues may also be a 2s suitable according to the invention.
If using other attachment groups, such as -SH groups, they may be added and/or removed analogously.
Substitution of the amino acid residues is preferred over insertion, as the impact on the 3-dimensional structure of the 3o polypeptide normally will be less pronounced.
The parent polypeptide In the context of the present invention, the term 35 "polypeptides" includes proteins, peptides and/or enzymes for pharmaceutical or industrial applications. Typically the polypeptides in question have a molecular weight in the range between about 1 to 1000 kDa, preferred 4 to 100 kDa, more preferred 12 to 60 kDa.
Pharmaceutical polypeptides The term "pharmaceutical polypeptides" is defined as polypep s tides, including peptides, such as peptide hormones, proteins and/or enzymes, being physiologically active when introduced into the circulatory system of the body of humans and/or animals.
Pharmaceutical polypeptides are potentially immunogenic as io they are introduced into the circulatory system.
Examples of "pharmaceutical polypeptides" contemplated according to the invention include insulin, ACTH, glucagon, somatostatin, somatotropin, thymosin., parathyroid hormone, pigmentary hormones, somatomedin, erythropoietin, luteinizing i5 hormone, chorionic gonadotropin, hypothalmic releasing factors, antidiuretic hormones, thyroid stimulating hormone, relaxin, interferon, thrombopoietin (TPO) and prolactin.
Industrial polypeptides 2o Polypeptides used for industrial applications often have an enzymatic activity. Industrial polypeptides (e.g. enzymes) are (in contrast to pharmaceutical polypeptides) not intended to be introduced into the circulatory system of the body.
It is not very like that industrial polypeptides, such as 2s enzymes used as ingredients in industrial compositions and/or products, such as detergents and personal care products, including cosmetics, come into direct contact with the circulatory system of the body of humans or animals, as such enzymes (or products comprising such enzymes) are not injected ao (or the like) into the bloodstream.
Therefore, in the case of the industrial polypeptide the potential risk is respiratory allergy (i.e. IgE response) as a consequence of inhalation of polypeptides through the respiratory passage.
35 In the context of the present invention "industrial polypep-tides" are defined as polypeptides, including peptides, proteins and/or enzymes, which are not intended to be administered to humans and/or animals.

Examples of such polypeptides are polypeptides, especially enzymes, used in products such as detergents, household article products, agrochemicals, personal care products, such as skin care products, including cosmetics and toiletries, oral and s dermal pharmaceuticals, composition use for processing textiles, compositions for hard surface cleaning, and compositions used for manufacturing food and feed etc.
Enzymatic activity io Pharmaceutical or industrial polypeptides exhibiting enzymatic activity will often belong to one of the following groups of enzymes including Oxidoreductases (E. C. 1, "Enzyme Nomenclature, (1992), Academic Press, Inc.), such as laccase and Superoxide dismutase (SOD); Transferases, (E.C. 2), such as is transglutaminases (TGases); Hydrolases (E. C. 3), including proteases, especially subtilisins, and lipolytic enzymes;
Isomerases (E. C. 5), such as Protein disulfide Isomerases (PDI).
Hydrolases Proteolytic enzymes Contemplated proteolytic enzymes include proteases selected from the group of Aspartic proteases, such as pepsins, Cysteine proteases, such as Papain, Serine~ proteases, such as subtilisins, or metallo proteases, such as Neutrase°.
2s Specific examples of parent proteases include PD498 (WO
93/24623 and SEQ ID NO. 2), Savinase° (von der Osten et al., (1993), Journal of Biotechnology, 28, p. 55+, SEQ ID NO 3), Proteinase K (Gunkel et al., (1989), Eur. J. Biochem, 179, p.
185-194), Proteinase R (Samal et al, (1990), Mol. Microbiol, 4, so p. 1789-1792), Proteinase T (Samal et al., (1989), Gene, 85, p.
329-333), Subtilisin DY (Betzel et al. (1993), Arch. Biophys, 302, no. 2, p. 499--502), Lion Y (JP 04197182-A), Rennilase~
(Available from Novo Nordisk A/S), JA16 (WO 92/17576), Alcalase°
(a natural subtilisin Carlberg variant) (von der Osten et al., 35 (1993), Journal of Biotechnology, 28, p. 55+), Subtilisin BPN' J. Mol. Biol. 178:389-413 (1984); Hirono S., Akagawa H., Mitsui Y., Iitaka Y. (Available from Novo Nordisk A/S).

Carbohydrases Parent carbohydrases may be defined as all enzymes capable of hydrolyzing carbohydrate chains (e. g. starches) of especially five and six member ring structures (i.e. enzymes classified under the Enzyme Classification number E.C. 3.2 (glycosidases) in accordance with the Recommendations (1992) of the Interna-tional Union of Biochemistry and Molecular Biology (IUBMB)). .
Examples include carbohydrases selected from those classified io under the Enzyme Classification (E. C.) numbers:
a-amylase (3.2.1.1) b-amylase (3.2.1.2), glucan 1,4-a-glucosidase (3.2.1.3), cellulase (3.2.1.4), endo-1,3(4)-b-glucanase (3.2.1.6), endo-1,4-b-xylanase (3.2.1.8), dextranase i5 (3.2.1.11), chitinase (3.2.1.14), polygalacturonase (3.2.1.15), lysozyme (3.2.1.17), b-glucosidase (3.2.1.21), a-galactosidase (3.2.1.22), b-galactosidase (3.2.1.23), amylo-1,6-glucosidase (3.2.1.33), xylan 1,4-b-xylosidase (3.2.1.37), glucan endo-1,3-b-D-glucosidase (3.2.1.39), a-dextrin endo-1,6-glucosidase zo (3.2.1.41), sucrose a-glucosidase (3.2.1.48), glucan endo-1,3-a-glucosidase (3.2.1.59), glucan 1,4-b-glucosidase (3.2.1.74), glucan endo-1,6-b-glucosidase (3.2.1.75), arabinan endo-1,5-a-arabinosidase (3.2.1.99), lactase (3.2.1.108), chitonanase (3.2.1.132).
Examples of relevant carbohydrases include a-1,3-glucanases derived from Trichoderma harzianum; a-1,6-glucanases derived from a strain of Paecilomyces; b-glucanases derived from Bacillus subtilis; b-glucanases derived from Humicola insolens;
3o b-glucanases derived from Aspergillus niger; b-glucanases derived from a strain of Trichoderma; b-glucanases derived from a strain of Oerskovia xanthineolytica; exo-1,4-a-D-glucosidases (glucoamylases) derived from Aspergillus niger; a-amylases derived from Bacillus subtilis; a-amylases derived from Bacillus amyloliquefaciens; a-amylases derived from Bacillus stearothermophilus; a-amylases derived from Aspergillus oryzae;
a-amylases derived from non-pathogenic microorganisms; a-galactosidases derived from Aspergillus niger; Pentosanases, xylanases, cellobiases, cellulases, hemi-cellulases deriver from Humicola insolens; cellulases derived from Trichoderma reesei;
cellulases derived from non-pathogenic mold; pectinases, cellulases, arabinases, hemi-celluloses derived from Aspergillus niger; dextranases derived from Penicillium lilacinum; endo-glucanase derived from non-pathogenic mold; pullulanases derived from Bacillus acidopullyticus; b-galactosidases derived from Kluyveromyces fragilis; xylanases derived from Trichoderma reesei;
to Specific examples of readily available commercial carbohydrases include Alpha-GalO, Bio-FeedO Alpha, Bio-FeedO Beta, Bio-Feedo Plus, Bio-FeedO Plus, Novozyme° 188, Carezyme°, Celluclast~, Cellusoft°, Ceremyl°, Citrozym0, Denimax0, Dezyme0, Dextrozyme0, is Finizym~, Fungamyl0, Gamanase0, Glucanex°, Lactozym~, Maltogenase0, Pentopan0, Pectinex0, Promozyme~, Pulpzyme0, Novamyl0, Termamyl°, AMG (Amyloglucosidase Novo), Maltogenase°, Aquazym~, Natalase~ (all enzymes available from Novo Nordisk A/S). Other carbohydrases are available from other companies.
zo It is to be understood that also carbohydrase variants are contemplated as the parent enzyme.
The activity of carbohydrases can be determined as described in 25 "Methods of Enzymatic Analysis", third edition, 1984, Verlag Chemie, Weinheim, vol. 4.
Oxidoreductases 3o Laccases Contemplated laccases include Polyporus pinisitus laccase (WO
96/00290), Myceliophthora laccase (WO 95/33836}, Schytalidium laccase (WO 95/338337), and Pyricularia oryzae Iaccase (Available from Sigma).
Peroxidase Contemplated peroxidases include B. pumilus peroxidases (WO
91/05858), Myxococcaceae peroxidase (WO 95/11964), Coprinus cinereus (WO 95/10602) and Arthromyces ramosus peroxidase {Kunishima et al. (1994), J. Mol. Biol. 235, p. 331-344).
Transferases s Transglutaminases Suitable transferases include any transglutaminases disclosed in WO 96/06931 (Novo Nordisk A/S) and WO 96/22366 (Novo Nordisk A/S) .
to Isomerases Protein Disulfide Isomerase Without being limited thereto suitable protein disulfide isomerases include PDIs described in WO 95/01425 (Novo Nordisk A/S).
is Contemplated isomerases include xylose/glucose Isomerase (5.3.1.5) including Sweetzyme~.
Lyases Suitable lyases include Polysaccharide lyases: Pectate lyases ao (4.2.2.2) and pectin lyases (4.2.2.10), such as those from Bacillus licheniformis disclosed in WO 99/27083.
The polymeric molecule 25 The polymeric molecules coupled to the polypeptide may be any suitable polymeric molecule, including natural and synthetic homo-polymers, such as polyols {i.e. poly-OH), polyamines (i.e.
poly-NHZ) and polycarboxyl acids (i.e. poly-COOH), and further hetero-polymers i.e. polymers comprising one or more different ao coupling groups e.g. a hydroxyl group and amine groups.
Examples of suitable polymeric molecules include polymeric molecules selected from the group comprising polyalkylene oxides (PAO), such as polyalkylene glycols (PAG), including polyethylene glycols (PEG), methoxypolyethylene glycols (mPEG) 3s and polypropylen glycols, PEG-glycidyl ethers (Epox-PEG), PEG-oxycarbonylimidazole (CDI-PEG), Branced PEGS, poly-vinyl alcohol (PVA), poly-carboxylates, poly-(vinylpyrolidone), poly-D,L-amino acids, polyethylene-co-malefic acid anhydride, polystyrene-co-malic acid anhydrid, dextrans including carboxymethyl-dextrans, heparin, homologous albumin, celluloses, including methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose carboxyethylcellulose and s hydroxypropylcellulose, hydrolysates of chitosan, starches such as hydroxyethyl-straches and hydroxy propyl-starches, glycogen, agaroses and derivates thereof, guar gum, pullulan, inulin, xanthan gum, carrageenin, pectin, alginic acid hydrolysates -and bio-polymers.
yo Preferred polymeric molecules are non-toxic polymeric molecules such as (m)polyethylene glycol ((m)PEG) which further requires a relatively simple chemistry for its covalently coupling to attachment groups on the enzyme's surface.
Generally seen polyalkylene oxides (PAO), such as is polyethylene oxides, such as PEG and especially mPEG, are the preferred polymeric molecules, as these polymeric molecules, in comparison to polysaccharides such as dextran, pullulan and the like, have few reactive groups capable of cross-linking.
Even though all of the above mentioned polymeric molecules zo may be used according to the invention the methoxypolyethylene glycols (mPEG) may advantageously be used. This arise from the fact that methoxyethylene glycols have only one reactive end capable of conjugating with the enzyme. Consequently, the risk of cross-linking is less pronounced. Further, it makes the zs product more homogeneous and the reaction of the polymeric molecules with the enzyme easier to control.
An example of a branched PEG conjugate is Branched PEG2-NHS-ester of Lysine (available from Shearwater).
Activation and couplina of c~olvmers to nolvneptides If the polymeric molecules to be conjugated with the polypeptide in question are not active, they must be activated by the use of a suitable technique. It is also contemplated according to the invention to couple the polymeric molecules to the polypeptide through a linker. Suitable linkers are well-known to the skilled person.

Methods and chemistry for activation of polymeric molecules as well as for conjugation of polypeptides are intensively described in the literature. Commonly used methods for activation of insoluble polymers include activation of s functional groups with cyanogen bromide, periodate, glutaraldehyde, biepoxides, epichlorohydrin, divinylsulfone, carbodiimide, sulfonyl halides, trichlorotriazine etc. (see R.F.
Taylor, (1991), "Protein immobilisation. Fundamental and applications", Marcel Dekker, N.Y.; S.S. along, (1992), to "Chemistry of Protein Conjugation and C:rosslinking", CRC Press, Boca Raton; G.T. Hermanson et al., (1993), "Immobilized Affinity Ligand Techniques", Academic Press, N.Y.). Some of the methods concern activation of insoluble polymers but are also applicable to activation of soluble polymers e.g. periodate, is trichlorotriazine, sulfonylhalides, divinylsulfone, carbodiimide etc. The functional groups being amino, hydroxyl, thiol, carboxyl, aldehyde or sulfydryl on the polymer and the chosen attachment group on the protein must be considered in choosing the activation and conjugation chemistry which normally consist ao of i) activation of polymer, ii) conjugation, and iii) blocking of residual active groups.
In the following a number of suitable polymer activation methods will be described shortly. However, it is to be understood that also other methods may be used.
a5 Coupling polymeric molecules to the free acid groups of po lypeptides may be performed with the aid of diimide and for e xample amino-PEG or hydrazino-PEG (Pollak et al., (1976), J.
Amr. Chem. Soc., 98, 289-291) or diazoacetate/amide (along et al., (1992), "Chemistry of Protein Conjugation and 3o Crosslinking", CRC Press).
Coupling polymeric molecules to hydroxy groups are generally very difficult as it must be performed in water. Usually hydrolysis predominates over reaction with hydroxyl groups.
Coupling polymeric molecules to free sulfhydryl groups can 35 be reached with special groups like maleimido or the ortho pyridyl disulfide. Also vinylsulfone (US patent no. 5,414,135, (1995), Snow et al.) has a preference for sulfhydryl groups but is not as selective as the other mentioned.

Accessible Arginine residues in the polypeptide chain may be targeted by groups comprising two vicinal carbonyl groups.
Techniques involving coupling electrophilically activated PEGS to the amino groups of Lysines may also be useful. Many of s the usual leaving groups for alcohols give rise to an amine linkage. For instance, alkyl sulfonates, such as tresylates (Nilsson et al., (1984), Methods in Enzymology vol. 104, Jacoby, W. B., Ed., Academic Press: Orlando, p. 56-66; Nilsson et al., (1987), Methods in Enzymology vol. 135; Mosbach, K., Ed.; Aca-io demic Press: Orlando, pp. 65-79; Scouten et al., (1987), Methods in Enzymology vol. 135, Mosbach, K., Ed., Academic Press:
Orlando, 1987; pp 79-84; Crossland et al., (1971), J. Amr. Chem.
Soc. 1971, 93, pp. 4217-4219), mesylates (Harris, (1985), supra;
Harris et al., (1984), J. Polym. Sci. Polym. Chem. Ed. 22, pp is 341-352), aryl sulfonates like tosylates, and para-nitrobenzene sulfonates can be used.
Organic sulfonyl chlorides, e.g. Tresyl chloride, effectively converts hydroxy groups in a number of polymers, e.g. PEG, into good leaving groups (sulfonates) that, when 2o reacted with nucleophiles like amino groups in polypeptides allow stable linkages to be formed between polymer and polypeptide. In addition to high conjugation yields, the reaction conditions are in general mild (neutral or slightly alkaline pH, to avoid denaturation and little or no disruption zs of activity), and satisfy the non-destructive requirements to the polypeptide.
Tosylate is more reactive than the mesylate but also more un-stable decomposing into PEG, dioxane, and sulfonic acid (Zalipsky, (1995), Bioconjugate Chem., 6, 150-165). Epoxides may ao also been used for creating amine bonds but are much less reactive than the above mentioned groups.
Converting PEG into a chloroformate with phosgene gives rise to carbamate linkages to Lysines. This theme can be played in many variants substituting the chlorine with N-hydroxy 3s succinimide (US patent no. 5,122,614, (1992); Zalipsky et al., (I992), Biotechnol. Appl. Biochem., 15, p. 100-114; Monfardini et al., (1995), Bioconjugate Chem., 6, 62-69, with imidazole (Allen et al., (1991), Carbohydr. Res., 213, pp 309-319), with para-nitrophenol, DMAP (EP 632 082 A1, (1993), Looze, Y.) etc.
The derivatives are usually made by reacting the chloroformate with the desired leaving group. All these groups give rise to carbamate linkages to the peptide.
s Furthermore, isocyanates and isothiocyanates may be employed yielding ureas and thioureas, respectively.
Amides may be obtained from PEG acids using the same leaving groups as mentioned above and cyclic imid thrones (US patent no.
5,349,001, (1994), Greenwald et al.). The reactivity of these io compounds are very high but may make the hydrolysis to fast.
PEG succinate made from reaction with succinic anhydride can also be used. The hereby comprised ester group make the conju gate much more susceptible to hydrolysis (US patent no.
5,122,614, (1992), Zalipsky). This group may be activated with is N-hydroxy succinimide, Furthermore, a special linker can be introduced. The most commonly used is cyanuric chloride (Abuchowski et al., (1977}, J. Biol. Chem., 252, 3578-3581; US patent no. 4,179,337, (1979}, Davis et al.; Shafer et al., (1986), J. Polym. Sci. Polym. Chem.
2o Ed., 24, 375-378.
Coupling of PEG to an aromatic amine followed by diazotation yields a very reactive diazonium salt: which in situ can be reacted with a peptide. An amide linkage may also be obtained by reacting an azlactone derivative of PEG (US patent no.
2s 5,321,095, (1994}, Greenwald, R. B.) thus introducing an additional amide linkage.
As some peptides do not comprise many Lysines it may be advantageous to attach more than one PEG to the same Lysine.
This can be done e.g. by the use of 1,3-diamino-2-propanol.
3o PEGS may also be attached to the amino-groups of the enzyme with carbamate linkages (WO 95/11924, Greenwald et al.). Lysine residues may also be used as the backbone.
The coupling technique used in the examples is the N
succinimidyl carbonate conjugation technique descried in WO
35 90/13590 (Enzon) .
Method for preparina improved polypeptides It is also an object of the invention to provide a method for preparing improved polypeptides comprising the steps of:
a) identifying amino acid residues located on the surface of the 3-dimensional structure of the parent polypeptide in question, b) selecting target amino acid residues on the surface of said 3-dimensional structure of said parent polypeptide to be modified, c) substituting one or more amino acid residues selected in step b) with other amino acid residue, and/or' to d) coupling polymeric molecules to the amino acid residues in step b) and/or step c).
Step a) Identifyina amino acid residues located on the surface of the parent polypeptide 3-dimensional structure To perform the method of the invention a 3-dimensional structure of the parent polypeptide in question is required.
This structure may for example be an X-ray structure, an NMR
2o structure or a model-built structure. The Brookhaven Databank is a source of X-ray- and NMR-structures.
A model-built structure may be produced by the person skilled in the art if one or more 3-dimensional structures) exists) of homologous polypeptide(s) sharing at least 30%
a5 sequence identity with the polypeptide in question. Several software packages exist which may be employed to construct a model structure. One example is the Homology 95.0 package from MSI Inc.
Typical actions required for the construction of a model ao structure are: alignment of homologous sequences for which 3 dimensional structures exist, definition of Structurally Conserved Regions (SCRs), assignment of coordinates to SCRs, search for structural fragments/loops in structure databases to replace Variable Regions, assignment of coordinates to these as regions, and structural refinement by energy minimization.
Regions containing large inserts (>_3 residues) relative to the known 3-dimensional structures are known to be quite difficult to model, and structural predictions must be considered with care.
Having obtained the 3-dimensional structure of the polypeptide in question, or a model of the structure based on s homology to known structures, this structure serves as an essential prerequisite for the fulfillment of the method described below.
Step b) Selection of taraet amino acid residues io Target amino acid residues to be modified are according to the invention selected from those amino acid residues, wherein the Ca-atom is located less than 15 A from a ligand. In a preferred embodiment a possible Ca-atom should be closer to the ligand than the C°'-atom. In a more preferred embodiment the C°'-is atom of the amino acid residue is located less than 10 A from the ligand and Said amino acid residues have an accessibility of at least 15%, preferably at least 2U% and more preferably at least 30%.
2o Step c) Substitution Conservative substitution It is preferred to make conservative substitutions in the polypeptide when the polypeptide has to be conjugated, as as conservative substitutions secure that the impact of the substitution on the polypeptide structure is limited.
In the case of providing additional amino groups this may be done by substitution of Arginine to Lysine, both residues being positively charged, but only the Lysine having a free amino 3o group suitable as an attachment groups.
In the case of providing additional carboxylic acid groups the conservative substitution may for instance be an Asparagine to Aspartic acid or Glutamine to Glutamic acid substitution.
These residues resemble each other in size and shape, except 35 from the carboxylic groups being present on the acidic residues.

In the case of providing SH-groups the conservative substitution may be done by substitution of Threonine or Serine to Cysteine.
Which amino acids to substitute depends in principle on the s coupling chemistry to be applied.
When no coupling is performed after substitution there is in general no limit on the selection of amino acids for substitution. However, preferred amino acids for substitutions are substitutions to polar residues e.g. K, R, D, E, H, Q, N, S, to T, C. Also substitutions to residues with short side chains G
and A are preferred.
Further, when no coupling is to be performed, the changes may be in the form of addition or deletion of at least one amino acid for which the C°' atom is located within 15A from is the bound ligand, preferably deleting an amino acid.
Furthermore, the parent protein may be changed by substituting some amino acids and deleting/adding other.
Only substitutions which provide polypeptides with reduced immune response when evaluated in animal models are within the zo concept of the present invention.
The mutations) performed in step c) may be performed by standard techniques well known in the art, such as site-directed mutagenesis (see, e.g., Sambrook et al. (1989), Molecular Cloning. A Laboratory Manual, Cold Spring Harbor, NY.
zs A general description of nucleotide substitution can be found in e.g. Ford et al., 1991, Protein Expression and Purification 2, p. 95-107.
In a preferred embodiment of the invention, more than one amino acid residue is substituted, added or deleted, these 3o amino acids possibly being located close to different bound ' ligands. In that case, it may be difficult to assess a priori how well the functionality of the protein is maintained while antigenicity, immunogenicity and/or allergenicity is reduced.
This can be achieved by establishing a library of diversified 3s mutants each having one or more changed amino acids introduced and selecting those variants which show good retention of function and at the same time a good reduction in antigenicity.
In the case of protease, this can be tested by assaying the secreted variants for enzyme activity (as described below in the experimental section) and for antigen binding (e.g. by competitive ELISA using methods known in the art. (see e.g J.
Clausen, Immunochemical Techniques For. The Identification and s Estimation of Macromolecules, Elsevier, Amsterdam, 1988 pp.187-188). Specifically, the competivity ELISA can be performed with the wild-type protease coated on ELISA plates, and incubated with specific polyclonal anti-protease antiserum from rabbits in the presence of protease variant. The scope of these io embodiments of the invention is by no means limited to protease, which serves only to provide an example. A
diversified library can be established by a range of techniques known to the person skilled in the art (Reetz MT; Jaeger KE, in Biocatalysis - from Discovery to Application edited by Fessner is WD, Vol. 200, pp. 31-57 (1999); Stemmer, Nature, vol. 370, p.389-391, 1994; Zhao and Arnold, Proc. Natl. Acad. Sci., USA, vol. 94, pp. 7997-8000, 1997; or Yano et al., Proc. Natl. Acad.
Sci., USA, vol. 95, pp 5511-5515, 1998). In a more preferable embodiment, substitutions are found by a method comprising the zo following steps: 1) a range of substitutions, additions, and/or deletions are listed, 2) a library is designed which introduces a randomized subset of these changes in the amino acid sequence into the target gene, e.g. by random mutagenesis, 3) the library is expressed, and preferred variants are selected. In a zs most preferred embodiment, this method is supplemented with additonal rounds of screening and/or family shuffling of hits from the first round of screening (J. E. Ness, et al, Nature Biotechnology, vol. 17, pp. 893-896, 1999) and/or combination with other methods of reducing allergenicity by genetic means 30 (such as that disclosed in W092/10755).
Generation of site directed mutations Prior to mutagenesis the gene encoding the polypeptide of as interest must be cloned in a suitable vector. Methods for generating mutations in specific sites is described below.
Once the polypeptide-encoding gene has been cloned, desirable sites for mutation identified, and the residues) to WO 00/22103 PC'T/DK99/00542 substitute for the original ones) have been decided, these mutations can be introduced using synthetic oligonucleotides.
These oligonucleotides contain nucleotide sequences flanking the desired mutation sites; mutant nucleotides are inserted during s oligo-nucleotide synthesis. In a preferred method, Site-directed mutagenesis is carried out by SOE-PCR mutagenesis technique described by Kammann et al. (1989) Nucleic Acids Research 17(13), 5404, and by Sarkar G. and Sommer, S.S. (19.90);
Biotechniques 8, 404-407.
io Step d) Couplincs polymeric molecules to the optionally modified parent enzyme Polypeptide-polymer conjugates of the invention may be prepared by any coupling method known in the art including the is above mentioned techniques.
Preparation of enzyme variants Enzyme variants to be conjugated may be constructed by any suitable method. A number of methods are well established in the art. For instance enzyme variants according to the 2o invention may be generated using the same materials and methods described in e.g. WO 89/06279 (Novo Nordisk A/S), EP 130,756 (Genentech), EP 479,870 (Novo Nordisk A/S), EP 214,435 (Henkel), WO 87/04461 (Amgen), WO 87/05050 (Genex), EP appli-cation no. 87303761 (Genentech}, EP 260,105 (Genencor), WO
25 88/06624 (Gist-Brocades NV), WO 88/07578 (Genentech), WO
88/08028 (Genex), WO 88/08033 (Amgen), WO 88/08164 (Genex), Thomas et al. (1985) Nature, 318 375-3'76; Thomas et al. (1987) J. Mol. Biol., 193, 803-813; Russel and Fersht (1987) Nature 328 496-500.
Co~plinq of polymeric molecules to the~polypeptide in question See previous paragraphs Immunogenicity and Allergenicity "Immunogenicity" is a wider term than "antigenicity" and "allergenicity", and expresses the immune system's response to the presence of foreign substances. Said foreign substances are called immunogens, antigens and allergens depending of the type of immune response the elicit.
An "immunogen" may be defined as a substance which, when introduced into circulatory system of animals and humans, is s capable of stimulating an immunologic response resulting in formation of immunoglobulin.
The term "antigen"' refers to substances which by themselves are capable of generating antibodies when recognized as a non-self molecule.
io Further, an "allergen" may be defined as an antigen which may give rise to allergic sensitization or an allergic response by IgE antibodies (in humans, and molecules with comparable effects in animals).
is Assessment of immunoaencity Assessment of the immunogenicity may be made by injecting animal subcutaneously to enter the immunogen into the circulation system and comparing the response with the response of the corresponding parent polypeptide.
ao The "circulatory system" of the body of humans and animals means, in the context of the present invention, the system which mainly consists of the heart and blood vessels. The heart de-livers the necessary energy for maintaining blood circulation in the vascular system. The circulation system functions as the as organism' s transportation system, when the blood transports 02, nutritious matter, hormones, and other substances of importance for the cell regulation into the tissue. Further the blood removes COZ from the tissue to the lungs and residual substances to e.g. the kidneys. Furthermore, the blood is of importance for 3o the temperature regulation and the defence mechanisms of the body, which include the immune system.
A number of in vivo animal models exist for assessment of the immunogenic potential of polypeptides. Some of these models give a suitable basis for hazard assessment in man. Suitable models 3s include a mice model.
This model seek to identify the immunogenic response in the form of the IgG response in Balb/C mice being injected subcutaneously with modified and unmodified polypeptides.

Also other animal models can be used for assessment of the immunogenic potential.
A polypeptide having "reduced immunogenicity" according to the invention indicates that the amount of produced antibodies, s e.g. immunoglobulin in humans, and molecules with comparable effects in specific animals, which can lead to an immune response, is significantly decreased, when introduced into the circulatory system, in comparison to the corresponding parent polypeptide.
io For Balb/C mice the IgG response gives a good indication of the immunigenic potential of polypeptides.
Assessment of alleraenicity Assessment of allergenicity may be made by inhalation tests, is comparing the effect of intratracheally (into the trachea) administrated parent enzymes with the corresponding modified enzymes according to the invention.
A number of in vivo animal models exist for assessment of the allegenicity of enzymes. Some of these models give a suitable ao basis for hazard assessment in man. Suitable models include a guinea pig model and a mouse model. These models seek to identify respiratory allergens as a function of elicitation reactions induced in previously sensitised animals. According to these models the alleged allergens are introduced intratrach as eally into the animals.
A suitable strain of guinea pigs, the Dunkin Hartley strain, do not as humans, produce IgE antibodies in connection with the allergic response. However, they produce another type of anti-body the IgGlA and IgGlB (see e.g. Prentra, ATLA, 19, p. 8-14, ao 1991), which are responsible for their allergenic response to inhaled polypeptides including enzymes. Therefore, when using the Dunkin Hartley animal model, the relative amount of IgGlA
and IgGlB is a measure of the allergenicity level.
The Balb/C mice strain is suitable for intratracheal, as intredermal or subcutaneous exposure. Balb/C mice produce IgE as the allergic response.
More details on assessing respiratory allergens in guinea pigs and mice is described by Kimber et al.,(1996), Fundamental and Applied Toxicology, 33, p. 1-10.
Other animals such as rats, rabbits etc. may also be used for comparable studies.
s Composition The invention relates to a composition comprising a modified polypeptide of the invention.
The composition may be a pharmaceutical or industrial composition.
io The composition may further comprise other polypeptides, proteins or enzymes and/or ingredients normally used in e.g.
detergents, including soap bars, household articles, agrochemicals, personal care products, including skin care compositions, cleaning compositions for e.g. contact lenses, i5 oral and dermal pharmaceuticals, compasition use for treating textiles, compositions used for manufacturing food, e.g. baking, and food/feed etc.
Use of the polypepti~le 2o The invention also relates to the use of the method of the invention for reducing the immune response of polypeptides.
It is also an object of the invention to use the polypeptide-polymer conjugate or the polypeptide otherwise modified according to the invention to reduce the allergenicity 25 Of industrial products, such as detergents, such as laundry, disk wash and hard surface cleaning detergents, food or feed products, personal care products and textile products.
3o MATERIAh AND METHODS
Materials Enzymes:
PD498: Protease of subtilisin type shown in WO 93/24623. The sequence of PD498 is shown in SEQ ID NO. 1 and 2.
ss Savinase~: The sequence is shown in SEQ ID NO 3 (Available from Novo Nordisk A/S) Su~btilisin BPN': The sequence can be found in the SWISS-PROT
database. The sequence is also disclosed in:

GALLAGHER T., OLIVER J., BOTT R., BETZEL C., GILLILAND G.L.;
"Subtilisin BPN' at 1.6-A resolution: analysis for discrete disorder and comparison of crystal forms."; Acta Crystallogr. D
52:1125-1135(1996). The enzyme is available from Novo Nordisk s A/S.
Amylase AA560: The alkaline a-amylase may be derived from a strain of Bacillus sp. DSM 12649. The strain was deposited on 25t'' January 1999 by the inventors under the terms of the Budapest Treaty on the International Recognition of the Deposit io of Microorganisms for the Purposes of Patent Procedure at Deutshe Sammmlung von Microorganismen and Zellkulturen GmbH
(DSMZ), Mascheroder Weg lb, D-38124 Braunschweig DE. The sequence is shown in SEQ ID NO. 4.
i5 Strains:
B. subtilis 309 and 147 are variants of Bacillus Ientus, deposited with the NCIB and accorded the accession numbers NCIB
10309 and 10147, and described in iJS Patent No. 3,723,250 incorporated by reference herein.
2o E. coli MC 1000 (M.J. Casadaban and S.N. Cohen (1980); J.
Mol. Biol. 138 179-207), was made r-,m'~~ by conventional methods and is also described in US Patent Application Serial No.
039,298.
2s Vectors:
pPD498: E. coli - B. subtilis shuttle vector (described in US patent No. 5,621,089 under section 6.2.1.6) containing the wild-type gene encoding for PD498 protease (SEQ ID NO. 2). The same vector is use for mutagenesis in E. coli as well as for ao expression in B. subtilis.
35 Materials, chemicals and solutions:
Horse Radish Peroxidase labeled anti-rat-Ig (Dako, DK, P162, #
031; dilution 1:1000).

Mouse anti-rat IgE (Serotec MCA193; dilution 1:200).
Rat anti-mouse IgE (Serotec MCA419; dilution 1:100).
Biotin-labeled mouse anti-rat IgGl monoclonal antibody (Zymed 03-9140; dilution 1:1000) s Biotin-labeled rat anti-mouse IgGl monoclonal antibody (Serotec MCA336B; dilution 1:1000) Streptavidin-horse radish peroxidase (Kirkeg~rd & Perry 14-30-00; dilution 1:1000).
CovaLink NH2 plates (Nunc, Cat# 459439) io Cyanuric chloride (Aldrich) Acetone (Merck) Rat anti-Mouse IgGl, biotin (SeroTec, Cat# MCA336B) Streptavidin, peroxidase (KPL) Ortho-Phenylene-diamine (OPD) (Kem-en-Tec, Cat# 4260) is HzOz, 30% (Merck) Tween 20 (Merck) Skim Milk powder (Difco) HzS04 {Merck) ao Buffers and Solutions:
Carbonate buffer {0.1 M, pH 10 {1 liter)) Na2C03 10.60 g PBS (pH 7.2 (1 liter)) NaCl 8.00 g KCl 0.20 g K2HP0,, 1.04 g 2s KHZP04 0.32 g Washing buffer PBS, 0.05% (v/v) Tween 20 Blocking buffer PBS, 2% (wt/v) Skim Milk powder Dilution buffer PBS, 0.05% (v/v) Tween 20, 0.5% (wt/v) Skim Milk powder 3o Citrate buffer (O.1M, pH 5.0-5.2 (1 liter))NaCitrate 20.60 g Citric acid 6.30 g Sodium Borate, borax (Sigma) 3,3-Dimethyl glutaric acid (Sigma) CaClz (Sigma) 35 Tresyl chloride (2,2,2-triflouroethansulfonyl chloride) (Fluka) 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (Fluka) N-Hydroxy succinimide (Fluka art. 56480)) Phosgene (Fluka art. 79380) Lactose (Merck 7656) PMSF (phenyl methyl sulfonyl flouride) from Sigma Succinyl-Alanine-Alanine-Proline-Phenylalanine-para-nitroanilide (Suc-AAPF-pNP) Sigma no. S-7388, Mw 624.6 g/mole.
Activation of CovaLink plates:
~ Make a fresh stock solution of l0 mg cyanuric chloride per-ml acetone.
io ~ Just before use, dilute the cyanuric chloride stock solution into PBS, while stirring, to a final concentration of lmg/ml.
~ Add 100 ml of the dilution to each well of the CovaLink NH2 plates, and incubate for 5 minutes at room temperature.
~ Wash 3 times with PBS.
i5 ~ Dry the freshly prepared activated plates at 50°C for 30 minutes.
~ Immediately seal each plate with sealing tape.
~ Preactivated plates can be stored at room temperature for 3 weeks when kept in a plastic bag.
Test Animals:
Female Balb/C mice (about 20 grams) purchased from Bomholdtgaard, Ry, Denmark.
Female Brown-Norway rats, weighing on the average 180 g Eq,~ipment XCEL II (Novex) ELISA reader (Wmax, Molecular Devices) HPLC (Waters) ao PFLC (Pharmacia) Superdex-75 column, Mono-Q, Mono S from Pharmacia, SW.
SLT: Fotometer from SLT LabInstruments Size-exclusion chromatograph (Spherogel TSK-62000 SW).
Size-exclusion chromatograph (Superdex 200, Pharmacia, SW) Amicon Cell Enzymes for DNA manipulations Unless otherwise mentioned all enzymes for DNA
manipulations, such as e.g. restriction endonucleases, ligases etc., are obtained from New England Biolabs. Inc.
Mef~l.8 BPX: Composition (per liter) Potato starch 1008 Ground barley 50g io Soybean flour tog Na2HP04 X 12 Hz0 9g Pluronic O.lg Sodium caseinate 10g The starch in the medium is liquefied with a-amylase and is the medium is sterilized by heating at 120°C for 45 minutes.
After sterilization the pH of the medium is adjusted to 9 by addition of NaHC03 to 0.1 M.
20 Met~lOC;B
General molecular biology methods:
Unless otherwise mentioned the DNA manipulations and transformations were performed using standard methods of molecular biology (Sambrook et al. (1989) Molecular cloning: A
2s laboratory manual, Cold Spring Harbor lab., Cold Spring Harbor, NY; Ausubel, F. M. et al. (eds.) "Current protocols in Molecular Biology". John Wiley and Sons, 1995; Harwood, C. R., and Cutting, S. M. (eds.) "Molecular Biological Methods for Bacillus". John Wiley and Sons, 1990).
3o Enzymes for DNA manipulations were used according to the specifications of the suppliers.
Fermentation of PD498 variants Fermentation of PD498 variants in B. subtilis are performed 3s at 30°C on a rotary shaking table (300 r.p.m.) in 500 ml baffled Erlenmeyer flasks containing 100 ml BPX medium for 5 days. In order to make an e.g. 2 liter broth 20 Erlenmeyer flasks are fermented simultaneously.

Purification of PD498 variants Approximately 1.6 litres of PD498 variant fermentation broth are centrifuged at 5000 rpm for 35 minutes in 1 litre s beakers. The supernatants are adjusted to pH 7.0 using 10%
acetic acid and filtered on Seitz Supra S100 filter plates.
The filtrates are concentrated to approximately 400 ml using an Amicon CH2A OF unit equipped with an Amicon S1Y10 OF cartridge.
The OF concentrate is centrifuged and filtered prior to io absorption at room temperature on a Bacitracin affinity column at pH 7. The PD498 variant is eluted from the Bacitracin column at room temperature using 25% 2-propanol and 1 M sodium chloride in a buffer solution with 0.01 dime-thyl-glutaric acid, 0.1 M boric acid and 0.002 M calcium chloride adjusted to i5 pH 7.
The fractions with protease activity from the Bacitracin purification step are combined and applied to a 750 ml Sephadex G25 column (5 cm diameter) equilibrated with a buffer containing 0.01 dimethylglutaric acid, 0.1 M boric acid and Zo 0.002 M calcium chloride adjusted to pH 6Ø
Fractions with proteolytic activity from the Sephadex G25 column are combined and applied to a 150 ml CM Sepharose CL 6B
cat-ion exchange column (5 cm diameter) equilibrated with a buffer containing 0.01 M dimethylglutaric acid, 0.1 M boric as acid, and 0.002 M calcium chloride adjusted to pH 6Ø
The protease is eluted using a linear gradient of 0-0.5 M
sodium chloride in 1 litres of the same buffer.
Protease containing fractions from the CM Sepharose column are combined and filtered through a 2~ filter.
Determination of the molecule weight Electrophoretic separation of proteins was performed by standard methods using 4-20% gradient SDS poly acrylamide gels (Novex).
Proteins were detected by silver staining. The molecule weight 3s was measured relative to the mobility of Mark-12° wide range molecule weight standards from Novex.
Protease activity Analysis with Suc-Ala-Ala-Pro-Phe-pNa:
Proteases cleave the bond between the peptide and p-nitroaniline to give a visible yellow colour absorbing at 405 nm.
s Buffer: e.g. Britton and Robinson buffer pH 8.3 Substrate: 100 mg suc-AAPF-pNa is dissolved into 1 ml dimethyl sulfoxide (DMSO). 100 ml of this is diluted into 10 ml with Britton and Robinson buffer.
The substrate and protease solution is mixed and the to absorbance is monitored at 405 nm as a function of time and ~Saos nm/mln. The temperature should be controlled (20-50°C
depending on protease). This is a measure of the protease activity in the sample.
is Proteolytic Activity In the context of this invention proteolytic activity is expressed in Kilo NOVO Protease Units (KNPU). The activity is determined relatively to an enzyme standard (SAVINASE_), and the determination is based on the digestion of a dimethyl ao casein (DMC) solution by the proteolytic enzyme at standard conditions, i,e. 50°C, pH 8.3, 9 min. reaction time, 3 min.
measuring time. A folder AF 220/1 is available upon request to Novo Nordisk A/S, Denmark, which folder is hereby included by reference.
25 A GU is a Glycine Unit, defined as the proteolytic enzyme activity which, under standard conditions, during a 15-minutes' incubation at 40°C, with N-acetyl casein as substrate, produces an amount of NH2-group equivalent to 1 mmole of glycine.
Enzyme activity can also be measured using the PNA assay, 3o according to reaction with the soluble substrate succinyl alanine-alanine-proline-phenyl-alanine-para-nitrophenol, which is described in the Journal of American Oil Chemists Society, Rothgeb, T.M., Goodlander, B.D., Garrison, P.H., and Smith, L.A. , (1988) .
ELISA IcrE test system (for Brown Norway rats) A three layer sandwich ELISA is used to determine relative concentrations of specific antibodies.

The immunizing molecule is used as coating antigen with 10 mg per ml and 50 ml per well, in neutral phosphate buffer, incubated overnight at 4°C. All remaining binding spots on the well surface are blocked in 2 % skim milk, 200 ml per well in s phosphate buffer for at least 30 minutes at room temperature (RT) . All seras to be tested with this antigen are added at 50 ml per well to this plate using a 8-channel pipette in dilution series from 10 x diluted followed by 3-fold dilutions. Dilutions are made in phosphate buffer with 0.5 % skim milk and 0.05%
to Tween20, incubated 2 hours on agitation platform at RT. The "tracer" molecule is biotinylated Mouse anti Rat IgE 50 ml per well and diluted 2000 x in phosphate buffer with 0.5 % skim milk and 0.05% Tween 20, incubated 2 hours on an agitation platform at RT. Control (blank) was identical sequence but without rat i5 sera. 50 ml per well streptavidin horse raddish peroxidase, diluted 2000 x was incubated 1 hour on an agitation platform.
Colouring substrate at 50 ml per well is OPD (6 mg) and Hz02 (4 ml of a 30% solution) per 10 ml citrate buffer pH 5.2. The reaction is stopped using 100 ml per well 2 N HZS04. All 2o readings on SLT at 486 nm and 620 nm as reference. Data is calculated and presented in Lotus.
LISA procedure to determinerelative concentrations of IaE
antibodies in BALB/C mice 25 A three layer sandwich ELISA is used to determine relative concentrations of specific IgE serum antibodies.
1) Coat the ELISA-plate with 10 mg rat anti-mouse IgE or mouse anti-rat IgE/ml buffer 1.
50 ml/well. Incubate over night at 4°C.
ao 2) Empty the plates and block with Blocking buffer at least hour at room temperature.
200 ml/well. Shake gently. Wash the plates 3 times with Washing Buffer.
3)Incubate with mouse/rat sera, starting from undiluted and as continue with 2-fold dilutions. Keep some wells free for buffer 4 only (blanks). 50 ml/well.
Incubate for 30 minutes at room temperature. Shake gently. Wash the plates 3 times in Washing Buffer.

WO 00/22103 PC'T/DK99/00542 4) Dilute the enzyme in Dilution buffer to the appropriate protein concentration. 50m1/well.
Incubate for 30 minutes at room temperature. Shake gently. Wash the plates 3 times in Washing Buffer.
s 5) Dilute specific polyclonal anti-enzyme antiserum serum (pIg) for detecting bound antibody in Dilution buffer.
50m1/well. Incubate for 30 minutes at room temperature. Shake gently. Wash the plates 3 times in Washing Buffer.
6) Dilute Horseradish Peroxidase-conjugated anti-pIg-antibody io in Dilution buffer. 50 ml/well.
Incubate at room temperature for 30 minutes. Shake gently.
Wash the plates 3 times in Washing Buffer.
7 ) Mix 0 . 6 mg ODP/ml + 0 . 4 ~,1 H202/ml i.n substrate Buf fer . Make the solution just before use. Incubate for 10 minutes. 50 is ~1/well.
8)To stop the reaction: add Stop Solution. 50 ~1/well.
9 ) Read the plates at 492 nm with 620 nm as reference.
Data is calculated and presented in Lotus.
EXAMPLES
Example 1 is Subtilisin BPN' In order to identify the residues to be modified, a distance and a directional criteria are applied.
As disclosed earlier residues having their C"-atom closer 3o than 15 A to a ligand are targets for modification. Preferably, residues having their Ca-atom closer to the ligand bound than the C"-atom, thereby allowing a potential side chain to point in the direction of the ligand, are targets for modification.
The relevant distance can easily be measured using e.g.
ss molecular graphics programs like InsightII from Molecular Simulations INC.
Especially surface exposed residues, defined as having ACC>0 when applying the DSSP program to the relevant protein part of the structure, are targets for modification. The DSSP
program is disclosed in W. Kabsch and C. Sander, BIOPOLYMERS 22 (1983) pp. 2577-2637.
s In Thomas E. Creighton, PROTEINS; Structure and Molecular Priciples, WH Freeman and Company, NY, ISBN: 0-7167-1566-X
(1984) is disclosed a table listing the accessible surface areas of individual amino acid residues. In the table below l5%
and 20% accessibility has been determined.
io Total 20% of 15% of ACC Total Total AA AxA AxA

Ala 115 23,0 17,3 Arg 225 45,0 33,8 Asn 160 32,0 24,0 Asp 150 30,0 22,5 Cys 135 27,0 20,3 Gln 180 36,0 27,0 Glu 190 38,0 28,5 Gly 75 15,0 11,3 His 195 39,0 29,3 Ile 175 35,0 26,3 Leu 170 34,0 25,5 Lys 200 40,0 30,0 Met 185 37,0 27,8 Phe 210 42,0 31,5 Pro 145 29,0 21,8 Ser 115 23,0 17,3 Thr 140 28,0 21,0 Trp 255 51,0 38,3 Tyr 230 46,0 34,5 ~Val 155 31,0 23,3 When dividing the found accessible surface area (ACC) for each amino acid of the protein with the accessible surface area for that individual amino acid (found in the Creighton table) is the accessibility value in percent is obtained.
In order to find residues to modify, the method described above was applied to the X-ray structure of Subtilisin BPN' in complex with the inhibitor CI-2 (entry 2SNI in the Brookhaven Protein Data Bank).
ao Only the Subtilisin BPN' and the two metal ions in the WO 00/22103 ~PCT/DK99/00542 structure was used for the analysis. Both ions are calcium ions. They are found in site 1 and site 2.
The results of the analysis are seen in the tables below.
The columns shows the distance in A from the metal ion to the s C°'and Cp as well as the accessibility as determined by DSSP for each residue to modify.
Site 1:
resid res.no dist (C~')ACC ACC
dirt (Ca) (~) GLY 80 4.40 14 18.67 ASN 77 4.68 4.57 62 38.75 ASP 41 5.14 4.36 0 GLN 2 5.46 4.64 47 26.11 ALA 74 5.57 5.12 0 GLY 83 7.80 0 PRO 86 8.44 7.42 8 GLY 70 9.04 1 THR 208 9.38 8.66 0 HIS 39 10.41 9.97 3 PRO 5 10.46 10.17 18 12.41 LYS 43 10.62 10.53 137 68.50 TYR 214 10.68 9.62 75 32.61 GLN 206 11.79 11.27 88 48.89 VAL 8 12.42 10.89 2 THR 22 13.14 12.12 22 15.71 GLY 215 13.52 14 18.67 PRO 14 13.53 13.29 45 31.03 HIS 17 13.64 12.25 28 14.36 THR 66 13.80 13.76 0 SER 9 14.40 14.22 58 50.43 ALA 13 14.66 13.53 0 GLY 7 14.74 0 LEU 90 14.79 13.38 1 ASP 36 14.87 14.57 20 13.33 GLY 211 14.88 45 60.00 Site 2:

resid resno dist (C')dist (Cp)ACC ACC

( AxA ( ~ ) ) GLU 195 4.44 4.28 48 25.26 ALA 176 4.67 3.85 0 GLY 169 5.16 0 ASP 197 5.90 5.14 21 14.00 VAL 165 8.35 6.96 6 ALA 151 8.54 8.04 0 GLY 166 9.43 14 18.67 GLY 193 9.46 0 GLY 264 9.63 7 VAL 149 9.85 9.50 3 GLY 178 10.74 0 VAL 139 10.95 9.63 0 GLY 154 11.31 17 22.67 SER 163 11.34 10.12 29 25.22 ARG 247 11.35 10.32 47 20.89 LYS 265 11.66 11.35 76 38.00 GLN 251 11.74 10.57 26 14.44 SER 191 11.83 11.04 0 SER 224 12.34 12.02 0 VAL 143 12.36 10.91 41 26.45 MET 124 12.43 11.71 0 GLY 127 12.44 61 81.33 SER 260 12.47 12.12 72 62.61 GLY 131 12.69 29 38.67 VAL 227 13.37 11.90 0 ' THR 220 13.55 12.34 3 LEU 250 13.58 12.73 3 LEU 135 13.60 13.21 6 GLY 266 13.93 0 GLY 128 14.04 16 21.33 SER 190 14.12 14.09 0 ALA 142 14.13 13.36 0 WO 00/22103 .PCT/DK99/00542 ILE 122 14.17 13.65 0 ALA 223 14.44 13.70 0 ASN 243 14.50 13.94 21 13.13 ALA 200 14.63 14.15 0 The table below shows functional preferred substitutions in site 1 and 2 of the BPN~. For Gly 80 the substitution G to S/T
G to N/Q and G to K/D means that Glycine in position 80 may preferably be substituted with Serine/Threonine or Asparagine/Glutamine or Lysine/Aspartic acid.
SITE1 Subtilisin BPN' G ly-80 G to S /T G to N /Q G to KlD

Asn-77 N to D/E N to K1R N to A1C

GIn-2 Q to D/E Q to KIR Q to A/C

P ro-5 P to G /A P to C IS P to K /D

Lys-43 K to SIT/C K to D/E/R K to QIN

Tyr-214 Y to N/A Y to A/G/C Y to K/H

Gln-206 Q to D/E Q to K/R Q to A/C

Thr-22 T to KIR T to QIN/A T to D/E/C

G ly-215 G to S IT G to N /Q G to K /D

Pro-14 P to G/A P to C/S P to K/D

Ser-9 S to KIR S to /NIA S to DIE/C

G ly-21 G to S /T G to N /Q G to K /D

SITE2 Subtilisin _BP_N' ~~~

Glu-195 G to SlT G to N/Q G to KID

G ly-166 G to SIT G to N/Q G to KID

G ly-154 G to S lT G to N /Q G to KID

Ser-163 S to K/R S to Q/N/A S to D/E/C

Arg-247 R to K/H R to Q/N R to A/CIE

Lys-265 K to S/TIC K to D/ElR K to Q/N

Val-143 V to A/G!H V to Q/E/C V to T/SIK

Gly-127 G to S/T G to N/Q G to K/D

Ser-260 S to K/R S to Q/N/A S to D/E/C

G ly-131 G to S IT G to N /Q G to K!D

Gly-128 G to S/T G to N/Q G to K/D

Example 2 The 3-dimensional structure of PD 498 as determined by X-ray crystallography in Brookhaven Protein Data Bank (PDB) format The sequence which was used to elucidate the three-dimensional structure forming the basis for the present invention consists of the 280 amino acids derived from Bacillus s sp. PD498, NCIMB No. 40484 as disclosed in sequence ID No. 2.
The structure of PD498 was solved in accordance with the principle for X-ray crystallographic methods given in "X-Ray Structure Determination", Stout, G.K. and Jensen, L.H., John Wiley & Sons, inc. NY, 1989 and "Protein Crystallography" by io Blundell, T.L. and Johnson, L.N., Academic Press, London, 1990.
The structural coordinates for the solved crystal structure of PD 498 at 2.2 A resolution using the isomorphous replacement method are given in a standard PDB format (Brookhaven Protein Data Base) in Appendix 1. It is to be understood that Appendix 1 i5 forms part of the present application.
In Appendix 1 the amino acid residues of the enzyme are identified by three-letter amino acid code (capitalized letters) .
PD498 has three bound metal ions . Site 1 is equivalent to 2o site 1 in subtilisin BPN' and contains a calcium ion. Site 2 does not have an equivalent in subtilisin BPN' and contains a calcium ion. Site 3 is in the same region as the 2nd site in subtilisin BPN' and does here contain a sodium ion and a monopropylene glycol ligand.
2s Applying the above method disclosed in example 1 results in:
Site 1:
residue resno dist (C')dirt (Ca) ACC (P.xA) ACC (%) GLY 89 4.26 4 ASP 5 5.02 3.92 0 ASP 48 5.10 4.36 0 ASN 86 5.15 4.73 33 20.63 ALA 82 5.84 4.97 0 GLY 87 6.05 41 54.67 GLY 92 7.33 0 TYR 8 7.87 7.12 12 TYR 7 8.01 7.63 89 38.70 PRO 47 8.13 8.09 59 40.69 s PRO 3 8.61 7.55 9 GLY 78 8.69 0 THR 213 9.19 8.55 0 ARG 51 10.39 9.61 162 72.00 HIS 46 10.41 9.93 1 1oLYS 52 10.56 9.41 10 TYR 219 10.74 9.79 56 24.35 ALA 211 11.55 11.03 9 GLN 12 11.67 10.44 22 12.22 GLY 218 12.00 18 24.00 15ALA 10 12.35 12.15 65 56.52 TYR 11 12.46 12.00 121 47.45 VAL 53 13.30 13.18 18 11.61 PRO 15 13.52 12.10 0 ARG 28 13.77 12.93 103 45.78 2oILE 99 14.16 13.16 0 ASP 43 14.36 14.04 8 TRP 1 14.43 13.90 71 27.84 GLY 14 14.60 1 GLY 234 14.85 0 2sGLY 29 14.97 13 17.33 ao Site 2:
resid resno dirt (Ca) dist (Cp) ACC (AxA) ACC
(%) ASN 65 4.25 4.04 65 40.63 ASP 61 4.98 3.62 88 58.67 35 ASP 63 5.30 4.43 46 30.67 ASP 58 5.39 3.87 0 MET 67 5.53 5.42 42 22.70 ILE 60 7.09 6.76 48 27.43 ARG 103 7.67 6.23 4 GLY 41 8.03 1 LEU 69 8.99 8.35 114 67.06 5 GLY 56 10.02 2 LYS 55 10.15 9.43 115 57.50 ALA 101 11.02 10.20 0 TYR 44 11.83 11.14 35 15.22 GLY 73 13.18 0 1o ASN 45 13.57 13.14 114 71.25 GLY 119 13.62 0 GLY 111 13.75 36 48.00 GLY 71 13.78 4 SER 115 13.82 12.77 24 20.87 i5 GLY 109 13.90 32 42.67 THR 74 13.96 13.69 0 PRO 215 14.41 13.20 30 20.69 VAL 53 14.70 13.64 18 11.61 VAL 37 14.80 14.62 1 Site 3:
resid resno dist (C') dist (C~3) ACC (AxA) ACC
(%) 25 ALA 179 4.07 4.05 0 ALA 181 4.65 4.11 0 TRP 200 6.65 6.57 46 18.04 ASP 202 6.86 6.02 19 12.67 ALA 160 7.85 7.10 0 3o VAL 158 8.84 8.28 0 THR 170 9.23 8.58 65 46.43 VAL 148 10.12 8.77 0 LYS 268 10.74 9.64 108 54.00 ARG 250 11.05 10.04 30 13.33 35 GLY 183 11.15 2 GLY 198 11.37 8 TRP 152 11.64 10.35 35 13.73 LEU 133 11.65 10.63 0 GLU 254 11.66 10.63 15 7.89 GLY 136 11.84 39 52.00 TYR 269 12.12 11.37 45 19.57 s GLY 163 12.15 11 14.67 SER 229 12.16 11.65 0 LEU 144 13.01 12.62 2 ASN 196 13.01 12.00 1 VAL 232 13.12 11.71 0 to LEU 131 13.25 12.69 0 ILE 253 13.27 12.22 1 ALA 151 13.59 12.87 0 THR 225 13.88 12.66 1 ASN 246 14.04 13.33 17 10.63 15 GLY 270 14.22 0 ILE 249 14.51 14.36 4 ALA 228 14.65 14.00 0 SER 141 14.78 14.70 21 18.26 ALA 236 14.93 13.63 0 zo The table below shows the preferred functional substitutions in site 1, 2 and 3 of PD498.

Asn-86 N to D/E N to K/R N to A/C

Gly-87 G to S/T G to N/Q G to KID

Tyr-7 Y to N/Q Y to A/G/C Y to K/H

P ro-4 P to G /A P to C /S P to K

Arg-51 R to K/H R to QIN R to A/C/E

Tyr-219 Y to N/Q Y to AIGIC Y to K/H

G ly-218 G to S/T G to N /Q G to KID

Ala-10 A to N/Q A to K/R A to D/E

Tyr-11 Y to N/Q Y to A/G/C Y to K/H

Arg-28 R to K/H R to Q/N R to A/CIE

Trp-1 W to N/Q W to A/G/C W to KIH

G ly-29 G to S/T G to N /Q G to KID

WO 00/22103 .PCT/DK99/00542 Asn-65 N to DIE N to K/R N to A/C

A s p-61 D to N /Q D to K /H D to A
/G /C

A s p-6 D to N /Q D to K /H D to A

M et-6 M to A /G M to ~ /E M to T

I le-6 I to A /G I to Q /E I to T

Leu-69 L to A/G/H L to Q/E/C L to T/SIK

Lys-55 K to S/TIC K to D/E/R K to DIN

Tyr-44 Y to N/Q Y to A/GIC Y to K/H

Asn-45 N to DIE N to K/R N to A/C

G ly-1 G to S /T G to N IQ G to K

S er-1 S to K /R S to Q /N S to D

G ly-10 G to S /T G to N /Q G to K

P ro-21 P to G /A P to C /S P to K

SITE3 _ PD498 Trp-200 W to N/Q W to A/G/C W to K/H

Thr-170 T to K/R T to Q/N/A T to D/E/C

Lys-268 K to S/T/C K to D/E/R K to Q/N

Gly-136 G to SIT G to N/Q G to KID

Tyr-269 Y to N/Q Y to A/G/C Y to K/H

Ser-141 S to K/R S to Q/NIA S to D/E/C

Example 3 Savinase io For this example the X-ray structure entry 1SVN in the Brookhaven Protein Data Bank was used. This structure contains two metal ions. Site 1 contains a calcium ion and is at a position equivalent to site 1 in subtilisin BPN'. Site 2 contains a calcium ion at a position equivalent to site 2 in ' is subtilisin BPN' . In the following list a SEQUENTIAL numbering have been applied and NOT the numbering system used in the structure file.
Site 1:
resid resno dist (C") dist (Ca) ACC ACC~'~
(.x11) ( °s ) GLY 78 4.28 14 18.67 ASN 75 4,74 4.64 61 38.13 ASP 40 5.08 4.34 0 GLN 2 5.39 4.59 45 25.0 ALA 72 5.49 4.99 0 GLY 81 7.68 0 PRO 84 8.28 7.29 5 GLY 68 8.88 1 THR 202 9.19 8.67 0 HIS 38 10.40 9.89 13 PRO 5 10.47 10.26 14 9.66 ASN 42 10.55 10.50 94 58.75 TYR 208 10.72 9.76 65 28.26 GLN 200 11.75 11.39 82 45.56 ILE 8 12.10 10.58 3 PRO 14 12.91 12.63 49 33.79 THR 22 13.01 12.24 29 20.71 HIS 17 13.44 12.07 29 14.87 ALA 13 13.78 12.63 0 GLY 7 14.60 2 LEU 88 14.86 13.68 0 GLY 223 14.89 0 GLY 23 14.93 0 Site 2:
resid resno dist (Ca) dist (C~') ACC ACC
(AxA) (%) ALA 170 4.88 4.24 0 GLY 189 5.10 46 61.33 ASP 191 7.22 6.52 6 ALA 149 7.79 7.05 0 ILE 159 8.29 6.89 1 VAL 147 8.98 8.40 0 VAL 137 9.81 8.44 0 GLY 187 10.71 3 GLY 258 10.85 3 WO 00/22103 .PCT/DK99/00542 ARG 241 10.90 9.77 39 17.33 GLY 172 11.27 0 GLY 125 11.66 46 61.33 THR 141 11.72 10.47 20 14.29 LEU 122 11.73 10.70 0 GLY 152 11.96 8 LEU 133 12.29 11.70 3 GLN 1$5 12.41 11.63 14 7.74 THR 218 12.51 11.95 0 LYS 245 12.79 11.71 48 24.00 SER 259 12.93 12.67 35 30.43 ASN 237 13.34 12.53 22 13.75 ALA 120 13.49 13.00 0 THR 254 13.53 13.19 100 71.43 VAL 221 13.62 12.14 0 ALA 140 13.65 13.13 0 VAL 145 13.91 13.88 0 THR 214 14.00 12.84 2 GLY 157 14.11 42 56.00 LEU 244 14.27 13.26 0 ALA 217 14.97 14.17 0 The table below shows the preferred functional substitutions in site 1 and 2 of Savinase.
s SITE1 Savinase G ly-78 G to SIT G to N/Q G to K/D

Asn-75 N to DIE N to K/R N to A/C

G In-2 Q to D/E Q to K/R Q to A/C

Asn-42 N to D/E N to K/R N to A/C

Tyr-208 Y to N/Q Y to A/G/C Y to K/H

G In-200 Q to D /E Q to K/R Q to AIC

Pro-14 P to G/A P to C/S P to KID

Thr-22 T to K/R T to Q/N/A T to DIE/C

His-17 H to S/T/C H to DIE H to Q/N

S ITE2 Savinase Gly-189 G to S/T G to NIQ G to KID

Arg-241 R to K/H R to Q/N R to A/C/E

Gly-125 G to SIT G to N/Q G to K/D

Lys-245 K to S/T/C K to D/E/R K to QIN

Ser-259 S to K/R S to Q/N/A S to D/E/C

Thr-254 T to K/R T to Q/N/A T to D/E/C

Gly-157 G to S/T G to N/Q G to KID

Example 4 Amylase ~AA560) For this example the structure of AA560 has been found by homology modelling using the BAN/Termamyl a-amylase structure disclosed in WO 96/23874 which is hereby incorporated by io reference. This structure contains two metal ions. Both site 1 and 2 contain a calcium ion.
The example shows how a 3-dimensional structure determined by model building using coordinates from a homologous structure, can be used to identify residues of the ligand i5 binding site, which may be modified in order to reduce the immune response.
Applying the method disclosed above results in:
Site 1:
Res ACC ACC
~x,~ C ) TYR 58: CA 23 10.00 GLY 59:CA 4 ALA 60:CA 0 VAL 103:CA 0 VAL 104:CA 1 MET 105:CA 6 ASN 106:CA 1 HIS 107:CA 6 LYS 108:CA 14 GLY 109:CA 2 VAL 122:CA 3 PRO 124:CA 27 18.62 ASN 126: CA 28 17.50 ARG 127:CA 17 ASN 128: CA 107 66.88 THR 141:CA 0 TRP 159: CA 75 29.41 TYR 160: CA 96 41.74 HIS 161:CA 2 PHE 162:CA 0 ASP 163:CA 1 GLY 164:CA 0 VAL 165:CA 6 ASP 166:CA5 64 42.67 ILE 177:CA 12 TYR 178:CA 0 LYS 179: CA 27 13.50 PHE 180:CA 0 LYS 185: CA 36 18.00 GLY 186: CA 24 32.00 TRP 187: CA 27 10.59 ASP 188:CA 0 TRP 189: CA 136 53.33 GLU 190: CA 39 20.53 VAL 191:CA 0 ASP 192:CA 11 THR 193: CA 84 60.00 GLU 194: CA 88 46.32 ASN 195: CA 36 22.50 GLY 196: CA 27 36.00 ASN 197:CA $

TYR 198: CA 41 17.83 ASP 199:CA 1 TYR 200:CA 2 LEU 201: CA 50 29.41 MET 202:CA 72 38.92 TYR 203; CA 93 40.43 ALA 204:CA 2 ASP 205:CA 0 ILE 206:CA 4 ASP 207:CA 6 MET 208:CA 5 ASP 209:CA 74 49.33 HIS 210:CA 39 20.00 VAL 213:CA 0 VAL 214: CA 26 16.77 LEU 217:CA 4 ILE 235:CA 0 ASP 236:CA 15 ALA 237:CA 5 VAL 238:CA 0 LYS 239:CA 14 HIS 240:CA 13 ILE 241:CA 1 LYS 242: CA 44 22.00 TYR 243:CA 5 SER 244: CA 40 34.78 PHE 245:CA 10 THR 246:CA 0 ARG 247: CA 60 26.67 TRP 249:CA 0 ALA 265:CA 0 GLU 266: CA 17 8.95 PHE 267:CA 2 TRP 268: CA 27 10.59 site 2:
Res ACC ACC
() ASN296: CA 25 15.63 LEU297: CA 1 TYR298: CA 68 29.57 ASN299: CA 72 45.00 ALA300: CA 0 SER301: CA 0 LYS302: CA 117 58.50 SER303: CA 43 37.39 GLY304: CA 70 93.33 GLY305: CA 8 10.67 ASN306: CA 149 93.13 TYR307: CA 49 21.30 ASP308: CA 59 39.33 MET309: CA 0 ARG310: CA 143 63.56 GLN311: CA 99 55.00 ILE312: CA 3 PHE313: CA 17 8.10 ASN314: CA 76 47.50 GLU345: CA 73 38.42 TRP347: CA 89 38.70 PHE348: CA 2 LEU351: CA 2 ALA352: CA 0 TYR404: CA 32 13.91 LEU405: CA 35 20.59 ASP406: CA 78 52.00 HIS407: CA 69 35.38 HIS408: CA 100 51.28 ASN409: CA 31 19.38 ILE410: CA 19 10.86 ILE411: CA 0 GLY412: CA 0 ILE429: CA 0 4$
MET430: CA 5 SER431: CA 0 ASP432: CA 5 GLY433: CA 19 25.33 ALA434: CA 73 63.48 GLY435: CA 35 46.67 GLY436: CA 21 28.00 ASN437: CA 86 53.75 VAL474: CA 0 ASN475: CA 53 33.13 GLY476: CA 41 54.67 GLY477: CA 29 38.67 SER478: CA 18 15.65 VAL479: CA 2 The table below shows functional preferred substitutions in s site 1 and 2 of the amylase AA560. For ASN 126 the substitution N to D/E means that Asparagine in position 126 may preferably be substituted with Aspartic acid or CTlutamic acid, Lysine or Arginine, or Alanine or Cysteine.
to wnu,~ __ ~~,~d ~~or~

Site S~te2 ASN N to N to N toAlC t.YS 302 K to Kto K to 126 LyE WR S~T/C GYE GYN

ASN N to N to N to SER 303 S to S to S to 128 LYE K/R A!C KfR (mVA D~EIC

TRP W to W toA/C,~CW to ASN 306 N to N to N to 158 N!Q KIH LyE KI AIC
R

TYR Y to Y toAIC,~CYto TYR 307 Y to Y toAlC-JCYto WH
160 WQ K~Fi IWQ

ASP D to D to D toAlGIC ASP 308 D to D to D toAfGJC
166 MQ WH NrQ K~I-i LYS K to Kto K toGYN ARG 310 R to R to R toAIGE
185 S~T/C DrE K>Ei CYN

TRP W to W toAIC~CW to GW 311 Q to Q to Q to 189 N~Q WH LYE K!R AIC

G11J E to E to E to ASN 314 N to N to N to 190 f~YQ K~Fi AIC,~C La'E K/R AlC

ASP D to D to D toAIC-JC GLU 345 E to E to E toAIC,~C
209 f~YQ WH N~Q KIH

HIS H to H to H to TRf 347 W to W to W to 210 SJT/C D'E Cf~N I~YQ AIGJC K/H

VAL V toGYNV to V to ASP406 D to D to D toAfGrC
214 C,IAIC KIWD I~YQ KM

LYS K to K to K to HIS 407 H to H to H to 242 Srf/C (YE GYN SrT'/C LYE CYN

SER S to S to S to HIS 408 H to H to H to 244 KIR GYf~YA L~E/C S~T/C LYE CYN

ARG R to R to R toAlG/E ALA434 A to Ato Ato GYE
247 WH CirN f~YQ K~R

ASN 437 N to N to N to CYE K/R AIC

ASN 475 N to N to N to DVE K~R AIC

GLY 476 G to G to G to SrT N~C~ WD

SER 478 S to S to S to K/R C~~VA CYEJC

Example 5 Coniuaation of Savinase variant R241K with activated bis-PEG-228 mg of the Savinase variant was incubated in 50 mM Sodium Borate pH 9.5 with 510 mg of N-succinimidyl carbonate activated bis-PEG 1000 in a reaction volume of approximately 30 ml. The reaction was carried out at ambient temperature using magnetic io stirring while keeping the pH within the interval 9.0-9.5 by addition of 0.5 M NaOH. The reaction time was 2 hours. The reaction was stopped by adding 1M HC1 to a final pH of 6Ø
Reagent excess was removed by ultra filtration using a Filtron-Ultrasette and the final product stored at -20°C, in SO mM
is Sodium Borate, 150 mM NaCl, 1 mM CaCl2, 50% mono propylene glycol at H 6Ø
Compared to the parent enzyme, residual activity was close to 100% towards a peptide substrate (succinyl-Ala-Ala-Pro-Phe-p-nitro-anilide?.
Example 6 2s Coniuaation of Savinase variant R241K with activated bis-PEG-353 mg of the Savinase variant was incubated in 50 mM Sodium Borate pH 9.5 with 1621 mg of N~-succinimidyl carbonate 3o activated bis-PEG 2000 in a reaction volume of approximately 35 ml. The reaction was carried out at ambient temperature using magnetic stirring while keeping the pH within the interval 9.0-9.5 by addition of 0.5 M NaOH. The reaction time was 2 hours.
The reaction was stopped by adding 1M HC1 to a final pH of 6Ø
as Reagent excess was removed by ultra filtration using a Filtron-Ultrasette and the final product stored at -20°C, in 50 mM
Sodium Borate, 150 mM NaCl, 1 mM CaCl2, 50% mono propylene glycol at H 6Ø

WO 00/22103 'PCT/DK99/00542 Compared to the parent enzyme, residual activity was close to 100% towards a peptide substrate (succinyl-A1a-Ala-Pro-Phe-p-nitro-anilide).
s Example 7 Determination of IcrE levels in rats of R241KbPEG1000 and R241KbPEG2000 io Methods:
Sample Management: Each sample was diluted to 0.075 mg protein/ml, and aliquoted in 1.5 ml. These fractions were sent to the stables for storage at -20°C until use. Additionally, 100 ~1 of the respective fractions were stored in the lab-is freezer at -20°C for immunochemical analysis at the beginning, halfway and at the end of the study. For each immunization and each analysis a new fraction was taken.
Immunization: Twenty intratracheal immunizations were performed weekly with 100 ~tl 0.9% (wt/vol) NaCl (control 2o group), or 100 ~.l of the protein dilution mentioned before.
(group 5 unmodified R241K variant of Savinase, group 6 R241K-bis-S-PEG1000, and group 7 R241K-bis-S-PEG2000. Each group contained 10 rats. Blood samples (2 ml) were collected from the eye one week after every second immunization. Serum was z5 obtained by blood clothing, and centrifugation.
ELISA: Specific IgE levels were determined using the ELISA's specific for rat IgE. The sera were titrated at dilutions, starting from undiluted. Optical densities were measured at 492/620 nm.
3o The results are shown in figure 1 . As can be seen the IgE
levels of the conjugated savinase variants R241K are reduced compared to the savinase variant R241K.
35 Example 8 Determination of IgE levels in mice of savinase variants R2410s R241E, R241H and R241K.
Female Balb/c mice, 9 weeks of age were immunised subcutaneously for 20 consecutive weeks, with wild type s savinase, and with variants having single mutations in position 8241 (R241Q, R241E, R241H, R241K). Every other week, IgG1 and IgE serum levels were determined by ELISA.
Sample Management: Each sample was diluted to 0.010 mg protein/ml, and aliquoted in 1.5 ml. These fractions were sent io to the stables for storage at -20°C until use. Additionally, ~1 of the respective fractions were stored in the lab-freezer at -20°C for immunochemical analysis at the beginning, halfway and at the end of the study. For each immunization and each is analysis a new fraction was taken.
Immunization: Twenty subcutanuous immunizations were performed weekly with 100 ~,1 0.9% (wt/vol) NaCl (control group), or 100 ~.1 of the protein dilution mentioned before.
Thus, group 1 received wild type Savinase, group 2 (R241Q), 2o group 3 (R241H), group 4 (R241E), and group 5 (R241K). Each group contained 10 mice. Blood samples (100 ~1) were collected from the eye one week after every second immunization. Serum was obtained by blood clotting, and centrifugation.
ELISA: Specific IgG1 levels were determined using the 2s ELISA
specific for mouse IgGl. The sera were titrated at ~ dilutions, starting from 1:160.
Specific IgE levels were determined using the ELISAs specific for mouse IgE. The sera were titrated at ~ dilutions, so starting from undiluted. Optical densities were measured at 492/620 nm.
Statistical analysis: Differences between data sets were analysed by using nonparametric methods: the Kruskal-Wallis Test and the Dunn's Multiple Comparison Test.
3s The results are shown in f figure 2 . As can be seen the IgE
levels of the Savinase variants are significantly reduced.

PAGE INTENTIONALLY LEFT BLANK

The structure of PD498 as determined by X-ray crystallography in Brookhaven Protein Data Bank (PDB) format.
CRYST 45.070 67.090 90.00 P212121 81.100 90.00 90.00 SCALE1 0.02219 0.000000.00000 0.00000 SCALE2 0.00000 0.014910.00000 0.00000 10SCALE3 0.00000 0.000000.01233 0.00000 ATOM 1 N TRPA L 17.560-14.24147.7421.00 15.33 ATOM 2 CA TRPA 1 18.953-13.78447.4871.00 15.36 ATOM 3 C TRPA L 19.164-12.34948.0021.00 14.46 ATOM 4 O TRPA 1 18.277-11.56747.6541.00 17.10 S

15ATOM 5 CB TRPA 1 19.316-13.77746.0001.00 21.00 ATOM 6 CG TRPA 1 20.729-13.51945.6071.00 15.22 ATOM 7 CD1 TRPA 1 21.877-14.24145.8451.00 14.54 ATOM 8 CD2 TRPA 1 21.184-12.39044.8571.00 16.51 ATOM 9 NE1 TRPA 1 22.998-13.64345.2451.00 18.87 20ATOM 10 CE2 TRPA 2 22.542-12.46944.6241.00 14.70 ATOM 11 CE3 TRPA 1 20.514-11w.27144.2721.00 20.67 ATOM 12 CZ2 TRPA 1 23.347-11.55943.9311.00 20.48 ATOM 13 CZ3 TRPA 1 21.309-10.38143.5961.00 16.65 ATOM 14 CH2 TRPA 1 22.661-10.47243.3601.00 16.74 25ATOM 15 N SERA 2 20.202-12.09348.8121.00 13.43 ATOM 16 CA SERA 2 20.289-10.69749.3121.00 15.64 ATOM 17 C SERA 2 21.710-10.24949.0141.00 15.52 ATOM 18 O SERA 2 22.776-10.60549.5011.00 18.28 ATOM 19 CB SERA 2 19.980-10.59150.8151.00 25.19 30ATOM 20 OG SERA :>. 18.701-11.13051.1191.00 27.27 ATOM 21 N PROA 3 21.785-9.317 48.0321.00 14.76 ATOM 22 CA PROA 3 23.056-8.803 47.5781.00 14.21 ATOM 23 C PROA 3 23.708-7.855 48.6061.00 14.51 ATOM 24 0 PROA :3 23.048-7.406 49.5561.00 14.63 35ATOM 25 CB PROA 3 22.743-8.050 46.2811.00 12.74 ATOM 26 CG PROA 3 21.293-7.620 46.4981.00 14.64 ATOM 27 CD PROA 3 20.663-8.776 47.2701.00 14.83 ATOM 28 N ASNA 4 25.005-7.718 48.4451.00 10.92 ATOM 29 CA ASNA 4 25.792-7.034 49.4771.00 13.99 40ATOM 30 C ASNA 4 25.899-5.526 49.3111.00 13.94 ATOM 31 O ASNA 4 26.667-4.870 50.0461.00 12.98 ATOM 32 CB ASNA 4 27.215-7.626 49.5021.00 12.72 ATOM 33 CG ASNA 4 28.075-7.328 48.3211.00 16.43 ATOM 34 OD1 ASNA 4 27.647-6.473 47.5091.00 14.80 45ATOM 35 ND2 ASNA 4 29.265-7.911 48.1551.00 18.33 ATOM 36 N ASPA 5 25.165-4.896 48.3601.00 11.85 ATOM 37 CA ASPA 5 25.401-3.474 48.1561.00 12.19 ATOM 38 C ASPA 5 25.065-2.624 49.3481.00 11.69 ATOM 39 O ASPA 5 23.954-2.816 49.9361.00 10.53 50ATOM 40 CB ASPA 5 24.570-2.988 46.9201.00 10.10 ATOM 41 CG ASPA 5 24.777-4.005 45.7801.00 9.83 ATOM 42 OD1 ASPA 5 24.199-5.106 45.7561.00 12.14 ATOM 43 OD2 ASPA 5 25.568-3.642 44.8711.00 12.15 ATOM 44 N PROA 6 25.900-1.745 49.7951.00 11.28 55ATOM 45 CA PROA 6 25.673-1.089 51.0841.00 11.29 ATOM 46 C PROA 6 24.481-0.190 51.1461.00 11.12 ATOM 47 O PROA 6 23.759-0.196 52.1801.00 12.14 ATOM 48 CB PROA 6 26.984-0.356 51.4261.00 12.53 ATOM 49 CG PROA 6 27.599-0.217 50.0141.00 14.20 60ATOM 50 CD PROA 6 27.226-1.453 49.2021.00 11.88 ATOM 51 N TYRA 7 24.1430.465 50.0461.00 11.91 ATOM 52 CA TYRA 7 23.0151.415 50.1371.00 12.11 ATOM 53 C TYRA 7 21.7330.635 49.8751.00 11.41 ATOM 54 0 TYRA 7 20.6421.099 50.1721.00 11.81 65ATOM 55 CB TYRA 7 23.2372.509 49.0781.00 13.43 ATOM 56 CG TYRA '7 24,3753.451 49.4071.00 16.52 ATOM 57 CD1 TYRA 7 24.8973.394 50.7321.00 19.41 ATOM 58 CD2 TYRA 7 24.900 4.310 48.5181.00 25.90 6 ATOM 59 CE1 TYRA 7 25.932 4.231 51.0781.00 23.70 6 ATOM 60 CE2 TYRA 7 25.942 5.152 48.8851.00 25.53 6 ATOM 61 CZ TYRA 7 26.454 5.099 50.1571.00 30.83 6 ATOM 62 OH TYRA 7 27.491 5.983 50.4001.00 33.22 8 ATOM 63 N TYRA 8 21.819 -0.575 49.3111.00 11.44 7 ATOM 64 CA TYRA 8 20.685 -1.490 49.2581.00 10.93 6 ATOM 65 C TYRA 8 20.237 -1.852 50.6981.00 10.97 6 ATOM 66 O TYRA 8 19.073 -1.666 51.0301.00 10.70 B

10ATOM 67 CB TYRA 8 20.975 -2.737 48.4311.00 11.45 6 ATOM 68 CG TYRA 8 19.938 -3.813 48.5471.00 8.84 6 ATOM 69 CD1 TYRA 8 18.683 -3.646 47.8941.00 10.94 6 ATOM 70 CD2 TYRA 8 20.110 -4.990 49.2591.00 10.83 6 ATOM 71 CE1 TYRA 8 17.705 -4.627 47.9831.00 10.64 6 15ATOM 72 CE2 TYRA 8 19.112 -5.961 49.3501.00 12.45 6 ATOM 73 CZ TYRA 8 17.902 -5.782 48.6781.00 12.68 6 ATOM 74 OH TYRA 8 16.908 -6.733 48.7891.00 13.25 8 ATOM 75 N SERA 9 21.247 -2.270 51.4601.00 10.94 7 ATOM 76 CA SERA 9 20.949 -2.660 52.8541.00 11.34 6 20ATOM 77 C SERA 9 20.483 -1.450 53.6561.00 9.76 6 ATOM 78 O SERA 9 19.549 -1.582 54.4651.00 11.69 8 ATOM 79 CB SERA 9 22.271 -3.179 53.4481.00 12.98 6 ATOM 80 OG SERA 9 21.986 -3.491 54.8401.00 14.32 8 ATOM 81 N ALAA 10 21.018 -0.283 53.4281.00 9.72 7 25ATOM 82 CA ALAA 10 20.805 0.860 54.3321.00 9.30 6 ATOM 83 C ALAA 10 19.596 1.655 53.9651.00 12.42 6 ATOM 84 O ALAA 10 18.883 2.230 54.7941.00 11.71 8 ATOM 85 CB ALAA 10 22.036 1.757 54.3631.00 12.70 6 ATOM 86 N TYRA 11 19.352 1.779 52.6211.00 12.14 7 30ATOM 87 CA TYRA 1:1 18.374 2.754 52.1881.00 11.21 6 ATOM 88 C TYRA 1:1 17.339 2.269 51.1771.00 12.10 6 ATOM 89 O TYRA 11 16.323 2.972 51.0181.00 11.98 8 ATOM 90 CB TYRA 11 19.141 3.914 51.4481.00 9.82 6 ATOM 91 CG TYRA 11 20.208 4.587 52.2931.00 11.77 6 35ATOM 92 CD1 TYRA 1:1 19.815 5.317 53.4191.00 15.20 6 ATOM 93 CD2 TYRA 11 21.541 4.493 51.9701.00 16.78 6 ATOM 94 CE1 TYRA 1:1 20.773 5.955 54.1961.00 17.43 6 ATOM 95 CE2 TYRA 1:1 22.494 5.125 52.7561.00 19.92 6 ATOM 96 CZ TYRA 1:1 22.084 5.837 53.8641.00 18.68 6 40ATOM 97 OH TYRA 11 23.095 6.450 54.6261.00 20.61 8 ATOM 98 N GLNA 12 17.490 1.075 50.5731.00 9.57 7 ATOM 99 CA GLNA 12 16.421 0.747 49.6221.00 10.23 6 ATOM 100 C GLNA 12 15.251 0.052 50.2601.00 9.88 6 ATOM 101 O GLNA 12 15.422 -0.710 51.2351.00 10.94 8 45ATOM 102 CB GLNA 12 16.984 -0.155 48.4711.00 10.30 6 ATOM 103 CG GLNA 12 17.846 0.612 47.4911.00 9.41 6 ATOM 104 CD GLNA 12 18.378 -0.293 46.3871.00 9.03 6 ATOM 105 OE1 GLNA 12 19.616 -0.526 46.4341.00 10.69 8 ATOM 106 NE2 GLNA 12 17.572 -0.688 45.4381.00 9.50 7 50ATOM 107 N TYRA 13 14.053 0.139 49.6391.00 8.12 7 ATOM 108 CA TYRA 13 12.931 -0.656 50.0061.00 8.04 6 ATOM 109 C TYRA 13 12.175 -1.201 48.7931.00 11.46 6 ATOM 110 4 TYRA 13 11.392 -2.128 48.9121.00 10.57 8 ATOM 111 CB TYRA 13 11.899 0.042 50.9151.00 8.33 6 55ATOM 112 CG TYRA 13 11.153 1.131 50.1811.00 8.81 6 ATOM 113 CD1 TYRA 13 9.906 0.863 49.6041.00 8.32 6 ATOM 114 CD2 TYRA 1:3 11.653 2.441 50.0971.00 8.79 6 ATOM 115 CE1 TYRA 13 9.192 1.837 48.9131.00 9.23 6 ATOM 116 CE2 TYRA 13 10.942 3.429 49.3811.00 7.86 6 60ATOM 117 CZ TYRA 13 9.687 3.114 48.8351.00 9.91 6 ATOM 118 OH TYRA 13 8.956 4.116 48.2101.00 10.94 8 ATOM 119 N GLYA 14 12.538 -0.722 47.5911.00 10.19 7 ATOM 120 CA GLYA 14 11.702 -1.114 46.4281.00 10.55 6 ATOM 121 C GLYA 14 11.732 -2.616 46.1341.00 11.11 6 65ATOM 122 O GLYA 14 10.681 -3.246 45.9671.00 11.34 8 ATOM 123 N PROA 15 12.909 -3.176 46.0301.00 8.71 7 ATOM 124 CA PROA 15 12.958 -4.630 45.8101.00 9.03 6 ATOM 125 C PROA 15 12.372 -5.426 46.9961.00 10.83 6 WO 00/22103 ~PC'T/DK99/00542 ATOM 126 O PROA 15 11.557 -6.333 46.7721.00 10.41 ATOM 127 CB PROA 15 14.480 -4.906 45.6831.00 10.96 ATOM 128 CG PROA 15 15.102 -3.549 45.3041.00 9.61 ATOM 129 CD PROA 15 14.229 -2.508 46.0301.00 9.57 5 ATOM 130 N GLNA 16 12.741 -5.038 48.2221.00 10.84 ATOM 131 CA GLNA 16 12.216 -5.795 49.3821.00 9.58 ATOM 132 C GLNA 16 10.677 -5.822 49.4201.00 9.41 ATOM 133 0 GLNA 16 10.047 -6.896 49.7111.00 12.05 ATOM 134 CB GLNA 16 12.784 -5.110 50.6531.00 9.56 10ATOM 135 CG GLNA 16 14.295 -5.237 50.7501.00 10.85 ATOM 136 CD GLNA 16 15.079 -4.045 50.3011.00 9.18 ATOM 137 OE1 GLNA 16 14.615 -3.357 49.3281.00 11.39 ATOM 138 NE2 GLNA 16 16.242 -3.776 50.8671.00 10.81 ATOM 139 N ASNA 17 10.073 -4.629 49.2151.00 10.52 15ATOM 140 CA ASNA 17 8.627 -4.532 49.3471.00 9.85 ATOM 141 C ASNA 17 7.909 -5.151 48.1851.00 11.20 ATOM 142 O ASNA 17 6.658 -5.131 48.2441.00 14.71 B

ATOM 143 CB ASNA 17 8.208 -3.046 49.5091.00 11.59 ATOM 144 CG ASNA 17 8.432 -2.520 50.9371.00 13.87 20ATOM 145 OD1 ASNA 17 9.226 -3.101 51.6581.00 13.47 ATOM 146 ND2 ASNA 17 7.687 -1.460 51.2591.00 12.64 ATOM 147 N THRA 18 8.566 -5.563 47.1281.00 10.29 ATOM 148 CA THRA 18 7.890 -6.216 45.9921.00 12.51 ATOM 149 C THRA 18 8.300 -7.680 45.9741.00 12.13 25ATOM 150 0 THRA 18 8.100 -8.386 44.9631.00 11.44 ATOM 151 CB THRA 18 8.244 -5.529 44.6591.00 9.68 ATOM 152 OG1 THRA 18 9.696 -5.431 44.5251.00 10.42 ATOM 153 CG2 THRA 18 7.591 -4.187 44.6241.00 12.66 ATOM 154 N SERA 19 8.884 -8.204 47.0781.00 10.54 30ATOM 155 CA SERA 19 9.287 -9.606 47.1401.00 12.19 ATOM 156 C SERA 19 10.334 -9.982 46.0791.00 10.66 ATOM 157 0 SERA 19 10.372 -11.14345.6091.00 11.55 ATOM 158 CB SERA 19 8.113 -10.59447.0581.00 15.77 ATOM 159 OG SERA 19 7.242 -10.31548.1791.00 14.40 35ATOM 160 N THRA 20 11.176 -9.000 45.7571.00 11.05 ATOM 161 CA THRA 20 12.179 -9.303 44.7311.00 9.03 ATOM 162 C THRA 20 13.341 -10.15945.2121.00 10.84 ATOM 163 O THRA 20 13.841 -10.97444.4621.00 12.64 ATOM 164 CB THRA 20 12.652 -8.005 44.0671.00 11.34 40ATOM 165 OG1 THRA 20 11.486 -7.307 43.5841.00 10.88 ATOM 166 CG2 THRA 20 13.563 -8.230 42.8671.00 13.97 ATOM 167 N PROA 21 13.788 -10.14046.4741.00 10.13 ATOM 168 CA PROA 21 14.814 -11.02846.9401.00 10.60 ATOM 169 C PROA 21 14.417 -12.50646.7491.00 11.26 45ATOM 170 O PROA 21 15.311 -13.25646.2701.00 13.61 ATOM 171 CB PROA 21 14.916 -10.70148.4671.00 10.30 ATOM 172 CG PROA 21 14.710 -9.205 48.3541.00 10.51 ATOM 173 CD PROA 21 13.477 -9.064 47.4641.00 10.93 ATOM 174 N ALAA 22 13.151 -12.86246.9381.00 12.78 50ATOM 175 CA ALAA 22 12.706 -14.24646.6391.00 13.32 ATOM 176 C ALAA 22 12.644 -14.48245.1231.00 14.57 ATOM 177 O ALAA 22 13.070 -15.57644.6791.00 15.26 ATOM 178 CB ALAA 22 11.317 -14.45447.2821.00 13.94 ATOM 179 N ALAA 23 12.273 -13.42544.3961.00 12.81 55ATOM 180 CA ALAA 23 12.317 -13.61442.9171.00 14.03 ATOM 181 C ALAA 23 13.716 -13.88042.4771.00 12.10 ATOM 182 0 ALAA 23 13.915 -14.57741.4251.00 12.81 ATOM 183 CB ALAA 23 11.712 -12.34142.2611.00 12.05 ATOM 184 N TRPA 24 14.752 -13.21742.9891.00 11.24 60ATOM 185 CA TRPA 24 16.128 -13.38742.5911.00 11.78 ATOM 186 C TRPA 24 16.654 -14.84042.8451.00 13.08 ATOM 187 0 TRPA 24 17.723 -15.10442.3051.00 15.03 ATOM 188 CB TRPA 24 16.971 -12.36643.3341.00 15.91 ATOM 189 CG TRPA 24 16.890 -10.94242.8981.00 12.46 65ATOM 190 CD1 TRPA 24 16.549 -10.48841.6561.00 11.81 ATOM 191 CD2 TRPA 24 17.146 -9.775 43.6731.00 12.89 ATOM 192 NE1 TRPA 24 16.584 -9.109 41.5901.00 11.12 ATOM 193 CE2 TRPA 24 16.965 -8.633 42.8531.00 11.48 ATOM 194 CE3 TRP A24 17.514 -9.55445.015 1.00 14.60 6 ATOM 195 CZ2 TRP A24 17.132 -7.30043.254 1.00 12.39 6 ATOM 196 CZ3 TRP A24 17.643 -8.26845.452 1.00 12.43 6 ATOM 197 CH2 TRP A24 17.501 -7.16444.601 1.00 11.45 6 ATOM 198 N ASP A25 15.955 -15.60243.660 1.00 16.22 7 ATOM 199 CA ASP A25 16.307 -17.01943.816 1.00 19.28 6 ATOM 200 C ASP A25 16.013 -17.75842.511 1.00 19.51 6 ATOM 201 0 ASP A25 16.674 -18.76642.232 1.00 21.06 8 ATOM 202 CB ASP A25 15.474 -17.59044.987 1.00 15.40 6 10ATOM 203 CG ASP A25 15.889 -17.05046.378 1.00 16.84 6 ATOM 204 ODl ASP A25 14.914 -16.93147.182 1.00 20.99 8 ATOM 205 OD2 ASP A25 17.069 -16.79846.451 1.00 18.83 8 ATOM 206 N VAL A26 15.083 -17.23441.717 1.00 16.85 7 ATOM 207 CA VAL A26 14.677 -17.88340.450 1.00 16.29 6 15ATOM 208 C VAL A26 15.490 -17.30739.300 1.00 14.78 6 ATOM 209 O VAL A26 16.049 -18.04438.463 1.00 16.30 8 ATOM 210 CB VAL A26 13.181 -17.68940.253 1.00 15.77 6 ATOM 211 CG1 VAL A26 12.727 -18.26038.878 1.00 16.26 6 ATOM 212 CG2 VAL A26 12.288 -18.24941.341 1.00 14.73 6 20ATOM 213 N THR A27 15.627 -15.97039.244 1.00 13.62 7 ATOM 214 CA THR A27 16.434 -15.39238.191 1.00 13.55 6 ATOM 215 C THR A27 16.969 -13.99438.562 1.00 13.11 6 ATOM 216 O THR A27 16.239 -13.32739.294 1.00 12.83 8 ATOM 217 CB THR A27 15.570 -15.26736.899 1.00 16.42 6 25ATOM 218 OG1 THR A27 16.481 -14.65235.997 1.00 20.55 8 ATOM 219 CG2 THR A27 14.260 -14.53837.127 1.00 15.35 6 ATOM 220 N ARG A28 18.167 -13.71538.082 1.00 13.95 7 ATOM 221 CA ARG A28 18.707 -12.37638.350 1.00 14.11 6 ATOM 222 C ARG A28 18.914 -11.56937.058 1.00 13.63 6 30ATOM 223 O ARG A28 19.518 -10.52237.077 1.00 13.70 8 ATOM 224 CB ARG A28 20.007 -12.48739.167 1.00 13.10 6 ATOM 225 CG AARGA28 19.786 -12.59240.676 0.50 16.44 6 ATOM 226 CD AARGA28 21.015 -13.22941.319 0,50 13.92 6 ATOM 227 NE AARGA28 21.173 -14.65340.989 0.50 20.11 7 35ATOM 228 CZ AARGA28 22.394 -15.19841.007 0.50 22.04 6 ATOM 229 NH1 AARGA28 23.372 -14.37041.347 0.50 16.08 7 ATOM 230 NH2 AARGA28 22.629 -16.45640.719 0.50 19.93 7 ATOM 231 CG BARGA28 19.609 -13.09440.526 0.50 12.85 6 ATOM 232 CD BARGA28 20.809 -13.39441.414 0.50 12.14 6 40ATOM 233 NE BARGA28 21.589 -14.47140.795 0.50 12.31 7 ATOM 234 CZ BARDA28 21.281 -15.74640.991 0.50 10.72 6 ATOM 235 NH1 BARGA28 20.289 -16.18341.754 0.50 13.92 7 ATOM 236 NH2 BARGA28 22.032 -16.67840.382 0.50 18.15 7 ATOM 237 N GLY A29 18.305 -12.01835.941 1.00 12.92 7 45ATOM 238 CA GLY A29 18.362 -11.29634.672 1.00 12.43 6 ATOM 239 C GLY A29 19.326 -12.01933.736 1.00 11.85 6 ATOM 240 O GLY A29 19.589 -13.20233.991 1.00 16.10 8 ATOM 241 N SER A330 19.705 -11.32532.693 1.00 11.08 7 ATOM 242 CA SER A330 20.543 -11.99331.666 1.00 12.22 6 50ATOM 243 C SER A330 21.461 -10.94331.078 1.00 13.35 6 ATOM 244 O SER A330 21.121 -9.88930.574 1.00 13.86 8 ATOM 245 CB SER A330 19.712 -12.58330.525 1.00 15.19 6 ATOM 246 OG SER A33U 20.650 -12.91729.463 1.00 17.47 8 ATOM 247 N SER A331 22.855 -11.29231.059 1.00 14.23 7 55ATOM 248 CA SER A331 23.828 -10.40630.487 1.00 13.69 6 ATOM 249 C SER A331 23.784 -10.22428.959 1.00 12.34 6 ATOM 250 O SER A331 24.497 -9.32228.490 1.00 19.86 8 ATOM 251 CB SER A331 25.268 -10.72530.898 1.00 18.58 6 ATOM 252 OG SER A331 25.541 -12.03730.388 1.00 22.11 8 60ATOM 253 N THR A332 22.962 -11.06728.421 1.00 11.83 7 ATOM 254 CA THR A332 22.892 -10.96926.958 1.00 15.40 6 ATOM 255 C THR A332 21.538 -10.41326.518 1.00 17.11 6 ATOM 256 0 THR A332 21.235 -10.37625.323 1.00 16.83 8 ATOM 257 CB THR A332 22.969 -12.36726.285 1.00 17.26 6 65ATOM 258 OG1 THR A332 22.107 -13.30526.854 1.00 22.90 8 ATOM 259 CG2 THR A332 24.448 -12.81526.411 1.00 22.90 6 ATOM 260 N GLN A333 20.861 -9.71627.512 1.00 13.07 7 ATOM 261 CA GLN A333 19.630 -8.97427.160 1.00 12.63 6 ATOM 262 C GLN A333 19.830 -7.523 27.6311.00 13.44 6 ATOM 263 O GLN A333 20.686 -7.227 28.4611.00 12.88 B

ATOM 264 CB GLN A333 18.420 -9.526 27.8621.00 10.82 6 ATOM 265 CG GLN A333 18.173 -10.96227.3601.00 12.25 6 ATOM 266 CD GLN A333 16.999 -11.64327.9661.00 11.24 6 ATOM 267 OE1 GLN A333 16.554 -11.30029.0741.00 13.22 8 ATOM 268 NE2 GLN A333 16.375 -12.65727.3261.00 16.24 7 ATOM 269 N THR A33 18.960 -6.642 27.1021.00 11.79 7 ATOM 270 CA THR A33 18.984 -5.248 27.5231.00 11.06 6 10ATOM 271 C THR A33 17.582 -4.748 27.9091.00 12.71 6 ATOM 272 0 THR A33 16.558 -5.185 27.3941.00 10.99 B

ATOM 273 CB THR A33 19.560 -4.307 26.4521.00 12.32 6 ATOM 274 OG1 THR A33 18.727 -4.350 25.2741.00 15.31 8 ATOM 275 CG2 THR A33 20.994 -4.701 26.0981.00 15.29 6 15ATOM 276 N VAL A34 17.606 -3.738 28.7811.00 9.33 7 ATOM 277 CA VAL A34 16.448 -2.906 29.0471.00 9.03 6 ATOM 278 C VAL A34 16.817 -1.513 28.5451.00 10.82 6 ATOM 279 O VAL A34 17.894 -1.011 28.9471.00 11.09 8 ATOM 280 CB VAL A34 16.075 -2.760 30.5251.00 11.19 6 20ATOM 281 CG1 VAL A34 14.975 -1.712 30.7311.00 12.05 6 ATOM 282 CG2 VAL A34 15.715 -4.091 31.0921.00 14.97 6 ATOM 283 N ALA A35 16.062 -0.937 27.6161.00 9.42 7 ATOM 284 CA ALA A35 16.349 0.401 27.1491.00 12.14 6 ATOM 285 C ALA A35 15.788 1.446 28.0861.00 9.96 6 25ATOM 286 O ALA A35 14.616 1.449 28.4081.00 11.17 8 ATOM 287 CB ALA A35 15.810 0.631 25.7241.00 11.54 6 ATOM 288 N VAL A36 16.662 2.296 28.5541.00 8.03 7 ATOM 289 CA VAL A3E. 16.307 3.428 29.4351.00 9.34 6 ATOM 290 C VAL A36 16.255 4.660 28.5371.00 9.42 6 30ATOM 291 O VAL A3E 17.290 5.187 28.1681.00 10.30 B

ATOM 292 CB VAL A36 17.253 3.591 30.6101.00 7.40 6 ATOM 293 CG1 VAL A36 16.943 4.884 31.3571.00 12.02 6 ATOM 294 CG2 VAL A36 17.063 2.359 31.5211.00 9.73 6 ATOM 295 N LEU A37 15.003 4.992 28.1811.00 8.33 7 35ATOM 296 CA LEU A37 14.774 6.125 27.2451.00 9.07 6 ATOM 297 C LEU A37 14.527 7.355 28.1001.00 9.81 6 ATOM 298 O LEU A37 13.443 7.499 28.6951.00 10.06 8 ATOM 299 CB LEU A37 13.552 5.776 26.3801.00 10.14 6 ATOM 300 CG LEU A37 13.933 4.661 25.3621.00 11.82 6 40ATOM 301 CD1 LEU A37 12.792 3.693 25.2831.00 16.87 6 ATOM 302 CD2 LEU A37 14.217 5.400 24.0431.00 15.43 6 ATOM 303 N ASP A38 15.548 8.192 28.2161.00 9.99 7 ATOM 304 CA ASP A38 15.523 9.245 29.2651.00 7.85 6 ATOM 305 C ASP A38 16.535 10.334 29.0141.00 10.38 6 45ATOM 306 O ASP A38 16.838 10.581 27.8101.00 10.43 8 ATOM 307 CB ASP A38 15.714 8.495 30.5881.00 10.08 6 ATOM 308 CG ASP A38 15.095 9.159 31.8061.00 9.45 6 ATOM 309 OD1 ASP A38 15.411 10.307 32.1151.00 11.41 B

ATOM 310 OD2 ASP A38 14.232 8.509 32.4241.00 11.17 8 50ATOM 311 N SER A39 17.006 11.024 30.0391.00 8.46 7 ATOM 312 CA SER A39 17.914 12.155 29.8351.00 9.33 6 ATOM 313 C SER A39 19.333 11.738 29.5831.00 10.03 6 ATOM 314 0 SER A39 20.239 12.604 29.5441.00 12.39 8 ATOM 315 CB SER A39 17.825 13.046 31.0761.00 10.11 6 55ATOM 316 OG SER A39 18.301 12.377 32.2341.00 11.06 8 ATOM 317 N GLY A40 19.611 10.436 29.4221.00 10.54 7 ATOM 318 CA GLY A40 20.956 9.937 29.2761.00 11.06 6 ATOM 319 C GLY A40 21.309 9.106 30.5041.00 11.64 6 ATOM 320 0 GLY A40 20.508 9.044 31.4151.00 12.31 B

60ATOM 321 N VAL A41 22.464 8.494 30.4781.00 11.73 7 ATOM 322 CA VAL A41 22.862 7.692 31.6771.00 10.62 6 ATOM 323 C VAL A41 24.328 8.026 31.8751.00 11.66 6 ATOM 324 O VAL A41. 25.086 7.934 30.9121.00 13.26 8 ATOM 325 CB VAL A41 22.679 6.202 31.4271.00 10.61 6 65ATOM 326 CG1 VAL A41. 23.194 5.368 32.6181.00 10.20 6 ATOM 327 CG2 VAL A41. 21.181 5.897 31.2361.00 11.88 6 ATOM 328 N ASP A42 24.763 8.180 33.1361.00 11.44 7 ATOM 329 CA ASP A42 26.196 8.426 33.4021.00 13.13 6 WO 00/22103 PC'f/DK99/00542 ATOM 330 C ASPA 42 26.852 7.060 33.461 1.0012.56 6 ATOM 331 O ASPA 42 26.966 6.459 34.518 1.0013.21 8 ATOM 332 CB ASPA 42 26.379 9.170 34.745 1.0013.74 6 ATOM 333 CG ASPA 42 27.857 9.433 35.018 1.0018.39 6 ATOM 334 OD1 ASPA 42 28.140 10.03436.082 1.0021.20 8 ATOM 335 OD2 ASPA 42 28.672 9.005 34.208 1.0012.32 8 ATOM 336 N TYRA 43 27.358 6.645 32.283 1.0011.92 7 ATOM 337 CA TYRA 43 27.980 5.350 32.095 1.0012.00 6 ATOM 338 C TYRA 43 29.429 5.271 32.714 1.0013.62 6 10ATOM 339 0 TYRA 43 29.966 4.179 32.656 1.0014.17 8 ATOM 340 CB TYRA 43 28.031 4.952 30.604 1.0014.19 6 ATOM 341 CG TYRA 43 28.532 6.090 29.719 1.0013.96 6 ATOM 342 CD1 TYRA 43 29.876 6.376 29.758 1.0021.10 6 ATOM 343 CD2 TYRA 43 27.649 6.817 28.949 1.0013.89 6 -15ATOM 344 CE1 TYRA 43 30.377 7.422 28.982 1.0023.78 6 ATOM 345 CE2 TYRA 43 28.162 7.877 28.152 1.0017.35 6 ATOM 346 CZ TYRA 43 29.499 8.136 28.197 1.0021.91 6 ATOM 347 OH TYRA 43 30.054 9.174 27.458 1.0025.51 8 ATOM 348 N ASNA 44 29.860 6.400 33.220 1.0014.70 7 20ATOM 349 CA ASNA 44 31.144 6.357 33.963 1.0015.55 6 ATOM 350 C ASNA 44 30.923 6.074 35.433 1.0014.58 6 ATOM 351 O ASNA.44 32.889 5.965 36.221 1.0014.97 8 ATOM 352 CB ASNA 44 31.874 7.711 33.833 1.0015.61 6 ATOM 353 CG ASNA 44 32.294 7.939 32.344 1.0013.41 6 25ATOM 354 OD1 ASNA 44 32.052 9.147 32.103 1.0022.06 8 ATOM 355 ND2 ASNA 44 32.766 6.911 31.724 1.0016.45 7 ATOM 356 N HISA 45 29.653 6.081 35.908 1.0013.02 7 ATOM 357 CA HISA 45 29.474 5.865 37.376 1.0011.13 6 ATOM 358 C HISA 45 29.917 4.462 37.727 1.0010.90 6 30ATOM 359 O HISA 45 29.653 3.499 37.064 1.0011.84 8 ATOM 360 CB HISA 45 27.929 5.959 37.618 1.0013.07 6 ATOM 361 CG HISA 45 27.519 6.069 39.068 1.0011.25 6 ATOM 362 NDl HISA 45 27.779 5.071 40.007 1.0011.49 7 ATOM 363 CD2 HISA 45 26.921 7.129 39.661 1.0010.98 6 35ATOM 364 CE1 HISA 45 27.307 5.517 41.159 1.0012.50 6 ATOM 365 NE2 HISA 45 26.810 6.732 41.035 1.0011.54 7 ATOM 366 N PROA 46 30.635 4.274 38.874 1.0011.14 7 ATOM 367 CA PROA 46 31.062 2.985 39.335 1.0011.47 6 ATOM 368 C PROA 46 29.978 1.921 39.380 1.0010.59 6 40ATOM 369 O PROA 46 30.196 0.767 39.040 1.0011.62 8 ATOM 370 CB PROA 46 31.677 3.220 40.742 1.0010.94 6 ATOM 371 CG PROA 46 32.043 4.671 40.670 1.0014.48 6 ATOM 372 CD PROA 46 31.085 5.353 39.688 1.0011.69 6 ATOM 373 N ASPA 47 28.728 2.326 39.705 1.0012.70 7 45ATOM 374 CA ASPA 47 27.682 1.314 39.825 1.0011.42 6 ATOM 375 C ASPA 47 26.899 1.133 38.520 1.0012.51 6 ATOM 376 O ASPA 47 25.902 0.416 38.521 1.0011.59 8 ATOM 377 CB ASPA 47 26.702 1.688 40.990 1.0012.16 6 ATOM 378 CG ASPA 47 26.587 0.469 41.896 1.009.76 6 50ATOM 379 OD1 ASPA 47 27.288 -0.54141.945 1.0010.00 8 ATOM 380 OD2 ASPA 47 25.518 0.471 42.653 1.0010.91 8 ATOM 381 N LEUA 4B 27.369 1.772 37.435 1.0010.44 7 ATOM 382 CA LEUA 48 26.697 1.562 36.155 1.0011.43 6 ATOM 383 C LEUA 48 27.672 1.221 35.027 1.0012.97 6 55ATOM 384 O LEUA 48 27.191 0.683 34.023 1.0011.53 8 ATOM 385 CB LEUA 48 25.972 2.837 35.638 1.0011.21 6 ATOM 386 CG LEUA 48 24.787 3.235 36.572 1.0010.61 6 ATOM 387 CD1 LEUA 48 24.254 4.643 36.324 1.0011.90 6 ATOM 388 CD2 LEUA 48 23.677 2.180 36.462 1.0013.84 6 60ATOM 389 N ALAA 49 28.975 1.461 35.252 1.0012.23 7 ATOM 390 CA ALAA 49 29.841 1.298 34.073 1.009.36 6 ATOM 391 C ALAA 49 29.855 -0.09133.513 1.0010.78 6 ATOM 392 O ALAA 49 30.048 -0.24532.236 1.0016.98 8 ATOM 393 CB ALAA 49 31.268 1.712 34.531 1.0012.23 6 65ATOM 394 N ARGA 50 29.747 -1.16434.276 1.0011.88 7 ATOM 395 CA ARGA 50 29.780 -2.52233.800 1.0011.57 6 ATOM 396 C ARGA 50 28.444 -2.94633.165 1.0012.53 6 ATOM 397 O ARGA 50 28.348 -4.04832.602 1.0016.06 8 ATOM 398 CB ARGA 50 30.103 -3.52434.930 1.0015.47 6 ATOM 399 CG ARGA 50 31.531 -3.24035.482 1.0011.83 6 ATOM 400 CD ARGA 50 32.055 -4.51336.187 1.0015.45 6 ATOM 401 NE ARGA 50 31.187 -4.89737.307 1.0016.23 7 ATOM 402 CZ ARGA 50 31.384 -5.96538.064 1.0019.96 6 ATOM 403 NH1 ARGA 50 32.429 -6.78237.837 1.0022.22 7 ATOM 404 NH2 ARGA 50 30.526 -6.23039.057 1.0018.50 7 ATOM 405 N LYSA 51 27.436 -2.07533.346 1.0011.50 7 ATOM 406 CA LYSA 51 26.104 -2.47132.907 1.0011.63 6 10ATOM 407 C LYSA 51 25.570 -1.74431.675 1.0012.77 6 ATOM 408 0 LYSA 51 24.582 -2.21231.104 1.0013.77 8 ATOM 409 CB LYSA 51 25.152 -2.12734.077 1.0012.63 6 ATOM 410 CG LYSA 51 25.387 -2.92235.380 1.0013.22 6 ATOM 411 CD LYSA 51 25.538 -4.41335.201 1.0014.71 6 -15ATOM 412 CE LYSA 51 24.312 -5.05134.628 1.0013.09 6 ATOM 413 NZ LYSA 51 23.056 -4.81535.491 1.0012.18 7 ATOM 414 N VALA 52 26.124 -0.62331.345 1.0011.71 7 ATOM 415 CA VALA 52 25.551 0.247 30.312 1.0010.53 6 ATOM 416 C VALA 52 26.166 0.046 28.941 1.0014.73 6 20ATOM 417 O VALA 52 27.383 0.061 28.778 1.0016.03 8 ATOM 418 CB VALA 52 25.711 1.692 30.750 1.0010.80 6 ATOM 419 CG1 VALA 52 25.233 2.601 29.613 1.0015.73 6 ATOM 420 CG2 VALA 52 24.874 1.987 32.005 1.0011.42 6 ATOM 421 N ILEA 53 25.247 -0.13027.987 1.0011.33 7 25ATOM 422 CA ILEA 53 25.609 -0.09826.552 1.0011.76 6 ATOM 423 C ILEA 53 25.210 1.272 26.028 1.0014.46 6 ATOM 424 0 ILEA 53 24.071 1.711 26.289 1.0013.75 8 ATOM 425 CB ILEA 53 24.877 -1.17925.791 1,0012.46 6 ATOM 426 CG1 ILEA 53 25.331 -2.53026.296 1.0014.67 6 30ATOM 427 CG2 ILEA 53 25.229 -1.05024.291 1.0011.24 6 ATOM 428 CD1 ILEA 53 24.535 -3.70225.780 1.0020.94 6 ATOM 429 N LYSA 54 26.112 1.975 25.367 1.0014.25 7 ATOM 430 CA LYSA 54 25.812 3.317 24.896 1.0012.81 6 ATOM 431 C LYSA 54 24.994 3.315 23.618 1.0012.98 6 35ATOM 432 O LYSA 54 25.458 2.835 22.572 1.0017.68 8 ATOM 433 CB LYSA 54 27.109 4.126 24.613 1.0013.38 6 ATOM 434 CG LYSA 54 27.905 4.467 25.886 1.0013.70 6 ATOM 435 CD LYSA 54 29.303 4.949 25.440 1.0023.57 6 ATOM 436 CE LYSA 54 30.311 4.482 26.488 1.0025.44 6 40ATOM 437 NZ LYSA 54 30.879 3.152 26.128 1.0039.20 7 ATOM 438 N GLYA 55 23.737 3.675 23.690 1.0012.13 7 ATOM 439 CA GLYA 55 22.853 3.848 22.554 1.0012.94 6 ATOM 440 C GLYA 55 22.968 5.304 22.070 1.0012.91 6 ATOM 441 O GLYA 55 23.771 6.146 22.479 1.0013.22 8 45ATOM 442 N TYRA 56 22.084 5.613 21.090 1.0013.51 7 ATOM 443 CA TYRA 56 22.092 6.918 20.449 1.0012.80 6 ATOM 444 C TYRA 56 21.594 8.052 21.346 1.0014.95 6 ATOM 445 O TYRA 56 20.699 7.845 22.158 1.0014.71 8 ATOM 446 CB TYRA 56 21.369 6.789 19.085 1.0012.24 6 50ATOM 447 CG TYRA 56 21.659 7.944 18.131 1.0012.40 6 ATOM 448 CD1 TYRA 56 22.915 7.978 17.542 1.0015.33 6 ATOM 449 CD2 TYRA 56 20.766 8.959 17.915 1.0013.06 6 ATOM 450 CE1 TYRA 56 23.255 9.010 16.664 1.0015.12 6 ATOM 451 CE2 TYRA 56 21.106 10.01717.016 1.0015.08 6 55ATOM 452 CZ TYRA 56 22.347 10.00216.421 1.0016.78 6 ATOM 453 OH TYRA 56 22.603 11.09715.574 1.0020.74 8 ATOM 454 N ASPA 57 22.042 9.257 21.061 1.0012.59 7 ATOM 455 CA ASPA 57 21.604 10.45621.735 1.0011.21 6 ATOM 456 C ASPA 57 20.882 11.31320.683 1.0014.11 6 60ATOM 457 O ASPA 57 21.559 11.87819.812 1.0014.49 8 ATOM 458 CB ASPA 57 22.814 11.20122.293 1.0015.01 6 ATOM 459 CG ASPA 57 22.480 12.52122.943 1.0013.70 6 ATOM 460 OD1 ASPA 57 21.400 13.01922.765 1.0013.31 8 ATOM 461 OD2 ASPA 57 23.391 13.05823.622 1.0017.36 8 65ATOM 462 N PHEA 58 19.554 11.26320.737 1.0011.17 7 ATOM 463 CA PHEA 58 18.729 12.00219.764 1.0013.01 6 ATOM 464 C PHEA 58 18.675 13.47520.055 1.0016.32 6 ATOM 465 0 PHEA 58 18.071 14.22219.282 1.0017.71 8 WO 00/22103 PCT%DK99/00542 ATOM 466 CB PHE 58 17.292 11.40719.790 1.0014.04 6 A

ATOM 467 CG PHEA 58 17.284 10.01819.247 1.0011.93 6 ATOM 468 CD1 PHEA 58 17.055 9.878 17.861 1.0012.43 6 ATOM 469 CD2 PHEA 58 17.546 8.841 19.950 1.0011.15 6 5 ATOM 470 CE1 PHEA 58 17.078 8.627 17.325 1.0013.41 6 ATOM 471 CE2 PHEA 58 17.564 7.606 19.383 1.0013.98 6 ATOM 472 CZ PHEA 58 17.345 7.456 17.990 1.0012.24 6 ATOM 473 N ILEA 59 19.092 13.94021.251 1.0013.50 7 ATOM 474 CA ILEA 59 19.180 15.35421.596 1.0015.65 6 10 ATOM 475 C ILEA 59 20.410 15.97420.964 1.0018.12 6 ATOM 476 O ILEA 59 20.220 17.01420.263 1.0023.97 8 ATOM 477 CB ILEA 59 19.241 15.47723.146 1.0015.66 6 ATOM 478 CG1 ILEA 59 17.951 15.10023.855 1.0019.46 6 ATOM 479 CG2 ILEA 59 19.590 16.92123.536 1.0022.80 6 15 ATOM 480 CD1 ILEA 59 16.626 15.69523.499 1.0021.33 6 ATOM 481 N ASPA 60 21.568 15.42121.177 1.0019.52 7 ATOM 482 CA ASPA 60 22.810 15.97420.563 1.0020.37 6 ATOM 483 C ASPA 60 23.051 15.391:19.1761.0023.56 6 ATOM 484 0 ASPA 60 24.039 15.84218.532 1.0021.66 8 20 ATOM 485 CB ASPA 60 24.011 15.63821.423 1.0024.04 6 ATOM 486 CG ASPA 60 24.163 16.25122.799 1.0029.23 6 ATOM 487 OD1 ASPA 60 23.498 17.27923.093 1.0027.86 8 ATOM 488 OD2 ASPA 60 24.968 15.67623.597 1.0022.41 8 ATOM 489 N ARGA 61 22.353 14.34118.772 1.0018.93 7 25 ATOM 490 CA ARGA 61 22.668 13.63917.519 1.0018.74 6 ATOM 491 C ARGA 61 24.106 13.18417.487 1.0022.32 6 ATOM 492 0 ARGA 61 25.042 13.41516.667 1.0020.82 8 ATOM 493 CB ARGA 61 22.241 14.56616.363 1.0020.03 6 ATOM 494 CG ARGA 61 20.743 14.75116.239 1.0027.06 6 30 ATOM 495 CD ARGA 61 20.210 15.68915.210 0.0020.00 6 ATOM 496 NE ARGA 61 19.042 16.30615.859 0.0020.00 7 ATOM 497 CZ ARGA 61 18.388 17.28815.185 0.0020.00 6 ATOM 498 NH1 ARGA 61 18.805 17.66613.981 0.0020.00 7 ATOM 499 NH2 ARGA 61 17.318 17.87215.746 0.0020.00 7 35 ATOM 500 N ASPA 62 24.436 12.34218.480 1.0018.53 7 ATOM 501 CA ASPA 62 25.742 11.75918.713 1.0019.28 6 ATOM 502 C ASPA 62 25.598 10.37819.351 1.0017.15 6 ATOM 503 0 ASPA 62 24.462 9.943 19.708 1.0017.25 8 ATOM 504 CB ASPA 62 26.663 12.71119.495 1.0019.71 6 40 ATOM 505 CG ASPA 62 26.330 12.88720.966 1.0025.11 6 ATOM 506 OD1 ASPA 62 25.880 11.94021.630 1.0017.54 8 ATOM 507 OD2 ASPA 62 26.480 13.99921.532 1.0026.53 8 ATOM 508 N ASNA 63 26.690 9.644 19.555 1.0016.07 7 ATOM 509 CA ASNA 63 26.714 8.291 20.046 1.0018.08 6 45 ATOM 510 C ASNA 63 27.071 8.238 21.540 1.0014.21 6 ATOM 511 O ASNA 63 27.589 7.220 22.004 1.0019.73 8 ATOM 512 CB ASNA 63 27.775 7.473 19.289 1.0022.90 6 ATOM 513 CG ASNA 63 29.335 7.776 19.315 0.0020.00 6 ATOM 514 OD1 ASNA 63 30.201 6.934 19.572 0.0020.00 8 50 ATOM 515 ND2 ASNA 63 29.600 9.079 19.152 0.0020.00 7 ATOM 516 N ASNA 64 26.975 9.391 22.182 1.0015.25 7 ATOM 517 CA ASNA 64 27.400 9.532 23.602 1.0016.89 6 ATOM 518 C ASNA 64 26.266 10.07924.433 1.0014.33 6 ATOM 519 0 ASNA 64 25.999 11.26224.588 1.0014.56 8 55 ATOM 520 CB ASNA 64 28.546 10.58623.613 1.0016.40 6 ATOM 521 CG ASNA 64 29.073 10.79725.019 1.0020.17 6 ATOM 522 OD1 ASNA 64 28.566 10.20025.964 1.0024.63 8 ATOM 523 ND2 ASNA 64 30.049 11.70225.183 1.0024.89 7 ATOM 524 N PROA 65 25.502 9.138 24.989 1.0011.21 7 60 ATOM 525 CA PROA 65 24.242 9.477 25.665 1.0013.27 6 ATOM 526 C PROA 65 24.437 9.812 27.151 1.0013.67 6 ATOM 527 O PROA 65 23.672 9.379 28.013 1.0012.46 8 ATOM 528 CB PROA 65 23.409 8.174 25.491 1.0012.03 6 ATOM 829 CG PROA 65 24.468 7.118 25.660 1.0013.48 6 530 CD PROA 65 25.668 7.692 24.820 1.0011.45 6 ATOM

ATOM 531 N META 66 25.496 10.59027.413 1.0012.49 7 ATOM 532 CA META 66 25.738 11.07428.772 1.0010.69 6 ATOM 533 C META 66 24.601 11.94729.263 1.0010.89 6 ATOM 534 O MET A 66 23.929 12.691 28.5601.00 12.73 8 ATOM 535 CB MET A 66 27.055 11.880 28.7601.00 14.19 6 ATOM 536 CG MET A 66 27.469 12.471 30.0791.00 13.14 6 ATOM 837 SD MET A 66 27.514 11.384 31.5421.00 14.24 16 ATOM 538 CE MET A 66 28.725 10.247 30.9601.00 16.73 6 ATOM 539 N ASP A 67 24.280 11.753 30.5411.00 12.39 7 ATOM 540 CA ASP A 67 23.223 12.441 31.2501.00 13.65 6 ATOM 541 C ASP A 67 23.622 13.848 31.7141.00 13.30 6 ATOM 542 0 ASP A 67 24.628 13.878 32.4571.00 14.71 8 10ATOM 543 CB ASP A 67 22.881 11.608 32.4981.00 12.61 6 ATOM 544 CG ASP A 67 21.584 12.025 33.1281.00 10.52 6 ATOM 545 ODlASP A 67 20.838 12.937 32.7661.00 10.71 8 ATOM 546 OD2ASP A 67 21.311 11.380 34.1941.00 11.95 8 ATOM 547 N LEU A 68 22.901 14.887 31.3981.00 12.91 7 15ATOM 548 CA LEU A 68 23.230 16.219 31.9351.00 11.29 6 ATOM 549 C LEU A 68 22.184 16.689 32.9381.00 17.10 6 ATOM 550 O LEU A 68 21.977 17.877 33.1911.00 18.88 8 ATOM 551 CB LEU A 68 23.273 17.220 30.7841.00 13.93 6 ATOM 552 CG LEU A 68 24.425 16.942 29.8291.00 18.76 6 20ATOM 553 CD1LEU A 68 24.437 18.059 28.7801.00 20.21 6 ATOM 554 CD2LEU A 68 25.787 16.856 30.5161.00 23.79 6 ATOM 555 N ASN A 69 21.312 15.750 33.3111.00 14.50 7 ATOM 556 CA ASN A 69 20.183 16.121 34.1851.00 13.25 6 ATOM 557 C ASN A 69 20.208 15.373 35.5071.00 15.28 6 25ATOM 558 O ASN A 69 20.055 15.947 36.5951.00 13.89 8 ATOM 559 CB ASN A 69 18.837 15.820 33.4931.00 13.49 6 ATOM 560 CG ASN A 69 17.700 16.122 34.4121.00 13.28 6 ATOM 561 OD1ASN A 69 17.292 15.258 35.2201.00 15.35 8 ATOM 562 ND2ASN A 69 17.132 17.347 34.4131.00 12.65 7 30ATOM 563 N GLY A 70 20.371 14.062 35.3891.00 11.32 7 ATOM 564 CA GLY A 70 20.428 13.126 36.5011.00 12.47 6 ATOM 565 C GLY A 70 19.248 12.180 36.5651.00 12.10 6 ATOM 566 O GLY A 70 19.392 11.092 37.1531.00 11.37 8 ATOM 567 N HIS A 71 18.098 12.548 36.0331.00 10.92 7 35ATOM 568 CA HIS A 71 16.928 11.677 36.0641.00 12.16 6 ATOM 569 C HIS A 71 17.178 10.320 35.4251.00 10.47 6 ATOM 570 0 HIS A 71 16.936 9.246 36.0051.00 10.18 8 ATOM 571 CB HIS A 71 15.866 12.443 35.3031.00 12.10 6 ATOM 572 CG HIS A 71 14.491 11.898 35.2811.00 10.59 6 40ATOM 573 ND1HIS A 71 14.070 11.083 34.2221.00 9.81 7 ATOM 574 CD2HIS A 71 13.448 12.059 36.1371.00 11.36 6 ATOM 575 CE1HIS A 71 12.804 10.755 34.4811.00 9.45 6 ATOM 576 NE2HIS A 71 12.394 11.339 35.6171.00 11.67 7 ATOM 577 N GLY A 72 17.747 10.309 34.2141.00 9.91 7 45ATOM 578 CA GLY A 72 17.985 9.019 33.5391.00 8.20 6 ATOM 579 C GLY A 72 18.943 8.130 34.2941.00 10.22 6 ATOM 580 0 GLY A 72 18.851 6.914 34.2101.00 11.25 B

ATOM 581 N THR A 73 19.996 8.710 34.9491.00 9.41 7 ATOM 582 CA THR A 73 20.943 7.870 35.6781.00 10.00 6 50ATOM 583 C THR A 73 20.264 7.190 36.9041.00 9.47 6 ATOM 584 O THR A 73 20.593 6.058 37.2151.00 10.68 8 ATOM 585 CB THR A 73 22.092 8.789 36.1401.00 12.41 6 ATOM 586 OGlTHR A 73 22.771 9.288 34.9421.00 11.16 B

ATOM 587 CG2THR A 73 23.170 8.096 36.9501.00 11.66 6 55ATOM 588 N HIS A 74 19.332 7.989 37.4891.00 9.10 7 ATOM 589 CA HIS A 74 18.615 7.468 38.6831.00 10.08 6 ATOM 590 C HIS A 74 17.725 6.317 38.2221.00 8.45 6 ATOM 591 O HIS A 74 17.755 5.193 38.7971.00 9.63 B

ATOM 592 CB HIS A 74 17.893 8.629 39.3361.00 11.77 6 60ATOM 593 CG HIS A 74 17.373 8.281 40.6971.00 10.65 6 ATOM 594 ND1HIS A 74 16.237 7.546 40.8921.00 10.09 7 ATOM 595 CD2HIS A 74 17.889 8.640 41.9091.00 10.65 6 ATOM 596 CE1HIS A 74 16.057 7.418 42.1941.00 10.71 6 ATOM 597 NE2HIS A 74 17.011 8.091 42.8471.00 11.39 7 65ATOM 598 N VAL A 75 16.991 6.560 37.1281.00 8.72 7 ATOM 599 CA VAL A 75 16.102 5.516 36.6011.00 9.96 6 ATOM 600 C VAL A 75 16.861 4.281 36.1961.00 9.80 6 ATOM 601 O VAL A 75 16.493 3.159 36.5691.00 9.73 8 ATOM 602 CB VAL A 75 15.368 6.143 35.3971.00 8.91 6 ATOM 603 CG1VAL A 75 14.632 5.037 34.6141.00 11.90 6 ATOM 604 CG2VAL A 75 14.373 7.207 35.9061.00 12.26 6 ATOM 605 N ALA A 76 18.045 4.463 35.5501.00 9.95 7 ATOM 606 CA ALA A 76 18.828 3.299 35.1091.00 10.80 6 ATOM 607 C ALA A 76 19.311 2.512 36.3251.00 8.05 6 ATOM 608 0 ALA A 76 19.350 1.268 36.2571.00 9.57 8 ATOM 609 CB ALA A 76 20.067 3.817 34.2961.00 11.31 6 ATOM 610 N GLY A 77 19.719 3.244 37.3941.00 8.67 7 10ATOM 611 CA GLY A 77 20.240 2.442 38.5091.00 10.66 6 ATOM 612 C GLY A 77 19.100 1.609 39.1541.00 8.38 6 ATOM 613 O GLY A 77 19.432 0.501 39.6281.00 9.24 8 ATOM 614 N THR A 78 17.898 2.146 39.1961.00 8.84 7 ATOM 615 CA THR A 78 16.820 1.294 39.7241.00 9.31 6 ~-15ATOM 616 C THR A 78 16.604 0.018 38.9301.00 7.59 6 ATOM 617 O THR A 78 16.379 -1.093 39.3961.00 10.79 8 ATOM 618 CB THR A 78 15.550 2.127 39.8331.00 9.05 6 ATOM 619 OG1THR A 78 15.760 3.175 40.7961.00 10.11 8 ATOM 620 CG2THR A 78 14.375 1.266 40.3271.00 10.42 6 20ATOM 621 N VAL A 79 16.642 0.189 37.5551.00 9.12 7 ATOM 622 CA VAL A 79 16.411 -0.985 36.6851.00 8.17 6 ATOM 623 C VAL A 79 17.466 -2.025.36.8751.00 8.18 6 ATOM 624 O VAL A 79 17.192 -3.196 36.9701.00 10.26 8 ATOM 625 CB VAL A 79 16.354 -0.577 35.1861.00 13.01 6 25ATOM 626 CG1VAL A 79 16.039 -1.862 34.3421.00 16.27 6 ATOM 627 CG2VAL A 79 15.250 0.388 34.8731.00 16.24 6 ATOM 628 N ALA A 80 18.753 -1.594 36.8611.00 9.76 7 ATOM 629 CA ALA A 80 19.799 -2.612 36.6791.00 9.95 6 ATOM 630 C ALA A 80 21.149 -2.084 37.0671.00 11.86 6 30ATOM 631 O ALA A 80 22.196 -2.334 36.4051.00 10.30 8 ATOM 632 CB ALA A 80 19.814 -3.107 35.1951.00 10.99 6 ATOM 633 N ALA A 81 21.287 -1.333 38.1721.00 10.11 7 ATOM 634 CA ALA A 81 22.606 -1.043 38.7381.00 8.94 6 ATOM 635 C ALA A 81 23.431 -2.334 38.8811.00 10.93 6 35ATOM 636 O ALA A 81 22.8?7 -3.375 39.0661.00 10.46 B

ATOM 637 CB ALA A 81 22.577 -0.382 40.1191.00 8.60 6 ATOM 638 N ASP A 82 24.767 -2.153 38.8971.00 9.68 7 ATOM 639 CA ASP A 82 25.667 -3.283 39.1951.00 13.33 6 ATOM 640 C ASP A 82 25.333 -3.770 40.6431.00 9.64 6 40ATOM 641 O ASP A B2 25.341 -2.891 41.4921.00 10.48 8 ATOM 642 CB ASP A 82 27.068 -2.744 39.0361.00 11.80 6 ATOM 643 CG ASP A 82 28.160 -3.765 38.8881.00 13.40 6 ATOM 644 ODlASP A 82 29.241 -3.350 38.3941.00 12.00 8 ATOM 645 OD2ASP A 82 27.952 -4.901 39.318I.00 12.74 8 45ATOM 646 N THR A 83 25.143 -5.049 40.7551.00 9.60 7 ATOM 647 CA THR A 83 24.598 -5.567 42.0411.00 10.59 6 ATOM 648 C THR A 83 25.509 -6.677 42.5741.00 11.19 6 ATOM 649 0 THR A 83 26.203 -7.421 41.8751.00 13.87 8 ATOM 650 CB THR A 83 23.240 -6.205 41.7151.00 12.73 6 50ATOM 651 OG1THR A 83 22.452 -5.144 41.1781.00 11.10 8 ATOM 652 CG2THR A 83 22.502 -6.829 42.9131.00 11.07 6 ATOM 653 N ASN A 84 25.558 -6.640 43.9261.00 11.01 7 ATOM 654 CA ASN A 84 26.421 -7.579 44.6721.00 12.42 6 ATOM 655 C ASN A 84 27.916 -7.260 44.4041.00 12.70 6 55ATOM 656 O ASN A 84 28.717 -8.171 44.5051.00 14.84 8 ATOM 657 CB ASN A 84 26.083 -9.024 44.3641.00 14.55 6 ATOM 658 CG ASN A 84 26.516 -9.910 45.5381.00 17.95 6 ATOM 659 OD1ASN A 84 26.308 -9.587 46.7121.00 16.26 8 ATOM 660 ND2ASN A 84 27.136 -11.03545.2211.00 19.28 7 60ATOM 661 N ASN A 85 28.181 -5.973 44.1371.00 11.37 7 ATOM 662 CA ASN A 85 29.540 -5.534 43.8831.00 11.85 6 ATOM 663 C ASN A 85 30.208 -4.745 44.9881.00 11.88 6 ATOM 664 O ASN A 85 31.195 -4.054 44.8631.00 14.14 8 ATOM 665 CB ASN A 85 29.614 -4.736 42.5741.00 12.33 6 65ATOM 666 CG ASN A 85 28.901 -3.418 42.6001.00 12.64 6 ATOM 667 OD1ASN A 85 27.959 -3.237 43.3651.00 11.43 8 ATOM 668 ND2ASN A 85 29.298 -2.439 41.7891.00 11.56 7 ATOM 669 N GLY A 86 29.539 -4.755 46.1631.00 12.16 7 ATOM 670 CA GLYA 86 29:982 -4.130 47.358 1.0012.38 6 ATOM 671 C GLYA 86 30.003 -2.614 47.377 1.0014.42 6 ATOM 672 O GLYA 86 30.591 -1.914 48.220 1.0018.31 8 ATOM 673 N ILEA 87 29.329 -2.004 46.388 1.0012.81 7 ATOM 674 CA ILEA 87 29.278 -0.603 46.104 1.0011.71 6 ATOM 675 C ILEA 87 27.805 -0.195 45.920 1.0010.49 6 ATOM 676 O ILEA 87 27.039 -0.898 45.250 1.0011.65 8 ATOM 677 CB ILEA 87 30.001 -0.268 44.734 1.0013.16 6 ATOM 678 CG1 ILEA 87 31.488 -0.601 44.998 1.0015.49 6 10ATOM 679 CG2 ILEA 87 29.743 1.152 44.317 1.0017.65 6 ATOM 680 CD1 ILEA 87 32.209 -0.655 43.631 1.0017.42 6 ATOM 681 N GLYA 88 27.452 0.954 46.442 1.0012.42 7 ATOM 682 CA GLYA 88 26.194 1.569 45.989 1.0011.25 6 ATOM 683 C GLYA 88 24.950 0.749 46.250 1.009.96 6 15ATOM 684 O GLYA 88 24.668 0.199 47.288 1.0011.18 8 ATOM 685 N VALA 89 24.106 0.667 45.193 1.0010.48 7 ATOM 686 CA VALA 89 22.741 0.149 45.260 1.008.53 6 ATOM 687 C VALA 89 22.639 -1.190 44.549 1.0011.23 6 ATOM 688 O VALA 89 23.666 -1.596 43.948 1.0010.26 8 20ATOM 689 CB VALA 89 21.727 1.175 44.689 1.0010.06 6 ATOM 690 CG1 VALA 89 21.615 2.398 45.639 1.0011.70 6 ATOM 691 CG2 VALA 89 22.081 1.654 43.263 1.0010.54 6 ATOM 692 N ALAA 90 21.477 -1.829 44.640 1.007.95 7 ATOM 693 CA ALAA 90 21.184 -3.046 43.869 1.008.13 6 25ATOM 694 C ALAA 90 20.078 -2.755 42.876 1.008.97 6 ATOM 695 O ALAA 90 19.085 -2.156 43.204 1.0010.80 8 ATOM 696 CB ALAA 90 20.696 -4.124 44.835 1.0011.49 6 ATOM 697 N GLYA 91 20.356 -3.266 41.655 1.009.50 7 ATOM 698 CA GLYA 91 19.313 -3.059 40.624 1.0010.29 6 30ATOM 699 C GLYA 91 18.278 -4.178 40.615 1.0010.12 6 ATOM 700 0 GLYA 91 18.457 -5.295 41.120 1.009.70 8 ATOM 701 N META 92 17.069 -3.849 40.120 1.0010.51 7 ATOM 702 CA META 92 15.995 -4.836 40.046 1.0010.40 6 ATOM 703 C META 92 16.312 -6.044 39.169 1.008.63 6 35ATOM 704 0 META 92 15.853 -7.173 39.474 1.0010.07 8 ATOM 705 CB META 92 14.700 -4.133 39.525 1.0010.66 6 ATOM 706 CG META 92 14.024 -3.346 40.670 1.0010.45 6 ATOM 707 SD META 92 13.253 -4.371 41.946 1.0011.55 16 ATOM 708 CE META 92 11.912 -5.093 41.007 1.0012.16 6 40ATOM 709 N ALAA 93 17.126 -5.840 38.098 1.008.87 7 ATOM 710 CA ALAA 93 17.598 -6.935 37.268 1.0010.60 6 ATOM 711 C ALAA 93 19.126 -6.945 37.275 1.0012.63 6 ATOM 712 O ALAA 93 19.803 -6.326 36.467 1.0011.07 8 ATOM 713 CB ALAA 93 17.041 -6.652 35.841 1.0011.23 6 45ATOM 714 N PROA 94 19.692 -7.579 38.287 1.0010.81 7 ATOM 715 CA PROA 94 21.127 -7.515 38.517 1.0012.42 6 ATOM 716 C PROA 94 21.963 -7.765 37.291 1.0012.53 6 ATOM 717 O PROA 94 22.990 -7.094 37.087 1.0014.34 8 ATOM 718 CB PROA 94 21.350 -8.634 39.578 1.0011.90 6 50ATOM 719 CG PROA 94 20.077 -8.538 40.360 1.0012.96 6 ATOM 720 CD PROA 94 18.941 -8.272 39.338 1.0012.20 6 ATOM 721 N ASPA 95 21.647 -8.786 36.456 1.0011.92 7 ATOM 722 CA ASPA 95 22.593 -9.148 35.399 1.0011.78 6 ATOM 723 C ASPA 95 22.215 -8.578 34.037 1.0012.73 6 55ATOM 724 0 ASPA 95 23.039 -8.643 33.127 1.0014.73 8 ATOM 725 CB ASPA 95 22.661 -10.67235.300 1.0012.28 6 ATOM 726 CG ASPA 95 23.335 -11.24236.572 1.0018.00 6 ATOM 727 ODl ASPA 95 24.147 -10.49637.098 1.0020.06 8 ATOM 728 OD2 ASPA 95 22.929 -12.38636.860 1.0027.51 8 60ATOM 729 N THRA 96 21.016 -7.956 33.957 1.0012.12 7 ATOM 730 CA THRA 96 20.613 -7.407 32.635 1.0012.08 6 ATOM 731 C THRA 96 21.336 -6.110 32.325 1.0011.03 6 ATOM 732 O THRA 96 21.555 -5.271 33.207 1.0012.04 8 ATOM 733 CB THRA 96 19.095 -7.274 32.590 1.0010.42 6 65ATOM 734 OG1 THRA 96 18.501 -8.585 32.725 1.0012.83 8 ATOM 735 CG2 THRA 96 18.523 -6.733 31.241 1.0011.86 6 ATOM 736 N LYSA 97 21.685 -5.929 31.026 1.0010.74 7 ATOM 737 CA LYSA 97 22.392 -4.675 30.685 1.0011.13 6 ATOM 738 C LYSA 97 21.400 -3.550 30.376 1.0012.38 ATOM 739 O LYSA 97 20.182 -3.832 30.148 1.0011.36 ATOM 740 CB ALYSA 97 23.198 -4.880 29.382 0.5012.99 ATOM 741 CG ALYSA 97 24.181 -6.046 29.425 0.5017.25 ATOM 742 CD A 97 25.152 -5.891 30.584 0.5015.11 ATOM 743 CE A 97 26.500 -6.533 30.211 0.5012.42 ATOM 744 NZ ALYSA 97 27.416 -6.547 31.382 0.5018.98 ATOM 745 CB BLYSA 97 23.436 -4.843 29.571 0.5014.58 ATOM 746 CG BLYSA 97 24.588 -5.769 29.995 0.5015.40 l0ATOM 747 CD BLYSA 97 25.597 -5.958 28.888 0.5016.62 ATOM 748 CE BLYSA 97 26.770 -6.845 29.293 0.5022.87 ATOM 749 NZ BLYSA 97 27.610 -6.168 30.320 0.5027.60 ATOM 750 N ILEA 98 21.861 -2.310 30.465 1.0010.38 ATOM 751 CA ILEA 98 21.048 -1.145 30.182 1.0010.34 15ATOM 752 C ILEA 98 21.459 -0.617 28.815 1.0010.92 ATOM 753 0 ILEA 98 22.618 -0.354 28.624 1.0012.98 B

ATOM 754 CB ILEA 98 21.342 -0.073 31.253 1.0010.78 ATOM 755 CG1 ILEA 98 20.779 -0.412 32.644 1.0011.69 ATOM 756 CG2 ILEA 98 20.758 1.269 30.847 1.0011.52 20ATOM 757 CD1 ILEA 98 21.604 0.253 33.746 1.0014.34 ATOM 758 N LEUA 99 20.522 -0.415 27.892 1.009.93 ATOM 759 CA LEUA 99 20.815 0.298 26.649 1.0010.06 ATOM 760 C LEUA 99 20.432 1.743 26.901 1.0010.51 ATOM 761 0 LEUA 99 19.225 2.085 27.071 1.009.69 25ATOM 762 CB LEUA 99 19.984 -0.359 25.506 1.0011.55 ATOM 763 CG LEUA 99 20.103 0.469 24.236 1.0011.50 ATOM 764 CD1 LEUA 99 21.553 0.416 23.750 1.0012.90 ATOM 765 CD2 LEUA 99 19.138 -0.039 23.204 1.0010.92 ATOM 766 N ALAA 100 21.356 2.645 27.040 1.0010.31 30ATOM 767 CA ALAA 100 21.060 4.046 27.338 1.009.78 ATOM 768 C ALAA 100 20.713 4.770 26.039 1.009.48 ATOM 769 O ALAA 100 21.557 4.833 25.119 1.0011.85 ATOM 770 CB ALAA 100 22.296 4.739 27.974 1.0011.54 ATOM 771 N VALA 101 19.480 5.268 26.012 1.009.16 35ATOM 772 CA VALA 101 19.062 6.013 24.795 1.0010.09 ATOM 773 C VALA 101 18.654 7.409 25.253 1.0010.39 ATOM 774 0 VALA 101 17.733 7.523 26.060 1.009.86 ATOM 7?5 CB VALA 101 17.937 5.305 24.085 1.0010.01 ATOM 776 CG1 VALA 101 17.556 6.021 22.742 1.0011.37 40ATOM 777 CG2 VALA 101 18.227 3.846 23.765 1.0012.69 ATOM 778 N ARGA 102 19.294 8.449 24.771 1.0010.96 ATOM 779 CA ARGA 102 19.041 9.797 25.252 1.0010.28 ATOM 780 C ARGA 102 18.003 10.499 24.396 1.0013.39 ATOM 781 0 ARGA 102 18.193 10.765 23.188 1.0014.69 45ATOM 782 CB ARGA 102 20.353 10.595 25.469 1.0011.23 ATOM 783 CG ARGA 102 19.993 12.026 25.927 1.0011.92 ATOM 784 CD ARGA 102 21.318 12.674 26.332 1.0011.26 ATOM 785 NE ARGA 102 21.088 13.998 26.872 1.0013.14 ATOM 786 CZ ARGA 102 21.537 15.160 26.462 1.0017.86 50ATOM 787 NH1 ARGA 102 22.286 15.196 25.353 1.0017.29 ATOM 788 NH2 ARGA 102 21.231 16.264 27.119 1.0013.78 ATOM 789 N VALA 103 16.871 10.780 24.968 1.0011.62 ATOM 790 CA VALA 103 15.757 11.445 24.301 1.0011.54 ATOM 791 C VALA 103 15.264 12.696 25.053 1.0012.58 55ATOM 792 O VALA 103 14.272 13.343 24.613 1.0015.25 ATOM 793 CB VALA 103 14.520 10.520 24.049 1.0012.23 ATOM 794 CG1 VALA 103 14.893 9.393 23.047 1.0015.01 ATOM 795 CG2 VALA 103 13.912 9.972 25.323 1.0014.01 ATOM 796 N LEUA 104 15.806 13.018 26.201 1.0013.17 60ATOM 797 CA LEUA 104 15.505 14.207 26.989 1.0013.95 ATOM 798 C LEUA 104 16.824 14.933 27.248 1.0011.06 ATOM 799 O LEUA 104 17.900 14.395 27.389 1.0012.49 ATOM 800 CB LEUA 104 14.908 13.887 28.361 1.0016.60 ATOM 801 CG LEUA 104 13.683 12.967 28.283 1.0012.90 65ATOM 802 CD1 LEUA 104 13.236 12.655 29.717 1.0015.91 ATOM 803 CD2 LEUA 104 12.590 13.532 27.433 1.0017.02 ATOM 804 N ASPA 105 16.640 16.297 27.188 1.0014.57 ATOM 805 CA ASPA 105 17.795 17.210 27.308 1.0011.73 ATOM 806 C ASP A105 18.165 17.49928.737 1.00 14.97 6 ATOM 807 0 ASP A105 17.755 16.82029.654 1.00 13.31 8 ATOM 808 CB ASP A105 17.447 18.49526.529 1.00 15.08 6 ATOM 809 CG ASP A105 16.415 19.37827.163 1.00 21.30 6 5 ATOM 810 OD1ASP A105 16.024 19.19928.320 1.00 16.65 8 ATOM 811 OD2ASP A105 15.940 20.34126.470 1.00 22.26 8 ATOM 812 N ALA A106 19.112 18.44228.926 1.00 15.23 7 ATOM 813 CA ALA A106 19.549 18.69130.304 1.00 13.88 6 ATOM 814 C ALA A106 18.448 19.14831.242 1.00 14.07 6 10ATOM 815 O ALA A106 18.632 18.97032.464 1.00 15.73 8 ATOM 816 CB ALA A106 20.623 19.79130.277 1.00 16.83 6 ATOM 817 N ASN A107 17.337 19.69430.787 1.00 15.93 7 ATOM 818 CA ASN A107 16.227 20.07631.629 1.00 18.54 6 ATOM 819 C ASN A107 15.139 19.03131.736 1.00 17.24 6 15ATOM 820 O ASN A107 14.093 19.27432.355 1.00 19.75 8 ATOM 821 CB ASN A107 15.587 21.34731.038 1.00 23.46 6 ATOM 822 CG ASN A10? 16.601 22.48131.051 1.00 26.78 6 ATOM 823 OD1ASN A107 17.162 22.75332.113 1.00 25.51 8 ATOM 824 ND2ASN A107 16.820 23.09929.904 1.00 26.59 7 20ATOM 825 N GLY A108 15.389 17.86331.134 1.00 16.72 7 ATOM 826 CA GLY A108 14.401 16.79331.185 1.00 19.02 6 ATOM 827 C GLY A108 13.346 16.91130.090 1.00 19.28 6 ATOM 828 O GLY A108 12.324 16.19930.201 1.00 23.96 8 ATOM 829 N SER A109 13.569 17.69529.071 1.00 18.97 7 25ATOM 830 CA SER A109 12.556 17.94128.046 1.00 19.93 6 ATOM 831 C SER A109 12.936 17.28126.738 1.00 19.26 6 ATOM 832 O SER A109 14.111 17.13226.434 1.00 16.92 8 ATOM 833 CB SER A109 12.425 19.45627.829 1.00 27.39 6 ATOM 834 OG SER A109 12.017 20.00829.076 1.00 36.14 8 30ATOM 835 N GLY A110 11.937 16.95025.927 1.00 19.72 7 ATOM 836 CA GLY A110 12.225 16.26224.673 1.00 20.18 6 ATOM 837 C GLY A110 11.058 16.41823.718 1.00 21.15 6 ATOM 838 O GLY A110 9.991 16.84824.138 1.00 27.11 S

ATOM 839 N SER A111 11.37? 16.28222.422 1.00 16.93 7 35ATOM 840 CA SER A111 10.303 16.41621.451 1.00 18.93 6 ATOM 841 C SER A111 9.655 15.05221.244 1.00 17.00 6 ATOM 842 O SER A111 10.258 14.00421.422 1.00 17.14 8 ATOM 843 CB SER A111 10.853 16.98220.148 1.00 21.62 6 ATOM 844 OG SER A111 11.640 16.03919.448 1.00 23.26 8 40ATOM 845 N LEU A112 8.354 15.12220.969 1.00 16.14 7 ATOM 846 CA LEU A112 7.698 13.80720.756 I.00 15.65 6 ATOM 847 C LEU A112 8.360 13.08319.577 1.00 14.33 6 ATOM 848 O LEU A112 8.393 11.83219.644 1.00 17.13 8 ATOM 849 CB LEU A112 6.187 13.94020.629 1.00 21.81 6 45ATOM 850 CG LEU A112 5.437 14.47021.857 1.00 22.13 6 ATOM 851 CD1LEU A112 3.926 14.46421.622 1.00 28.11 6 ATOM 852 CD2LEU A112 5.685 13.69923.153 1.00 25.27 6 ATOM 853 N ASP A113 8.726 13.76118.498 1.00 17.80 7 ATOM 854 CA ASP A113 9.300 12.97317.388 1.00 18.50 6 50ATOM 855 C ASP A113 10.622 12.34317.758 1.00 19.37 6 ATOM 856 O ASP A113 10.820 11.18717.316 1.00 18.50 8 ATOM 857 CB ASP A113 9.322 13.89416.160 1.00 20.24 6 ATOM 858 CG ASP A113 8.011 14.17315.519 1.00 19.59 6 ATOM 859 OD1ASP A113 7.995 15.14014.672 1.00 29.00 8 55ATOM 860 OD2ASP A113 6.943 13.58715.713 1.00 24.47 8 ATOM 861 N SER A114 11.438 12.96318.569 1.00 17.25 7 ATOM 862 CA SER A114 12.699 12.38519.032 1.00 20.51 6 ATOM 863 C SER A114 12.440 11.23019.998 1.00 17.38 6 ATOM 864 O SER A114 13.134 10.21219.896 1.00 16.74 8 60ATOM 865 CB SER A114 13.525 13.45919.733 1.00 25.71 6 ATOM 866 OG SER A114 14.016 14.31318.706 1.00 28.28 8 ATOM 867 N ILE A115 11.470 11.38020.891 1.00 13.42 7 ATOM 868 CA ILE A115 11.184 10.28321.816 1.00 11.54 6 ATOM 869 C ILE A115 10.687 9.106 21.001 1.00 11.78 6 65ATOM 870 O ILE A115 11.072 7.934 21.265 1.00 12.76 B

ATOM 871 CB ILE A115 10.132 10.72022.855 1.00 12.31 6 ATOM 872 CGlILE A115 10.815 11.77523.737 1.00 15.30 6 ATOM 873 CG2ILE A115 9.621 9.579 23.726 1.00 14.78 6 ATOM 874 CDl ILEA 115 9.771 12.522 24.5441.00 16.55 6 ATOM 875 N ALAA 116 9.807 9.353 20.0241.00 12.41 7 ATOM 876 CA ALAA 116 9.318 8.291 19.1781.00 11.46 6 ATOM 877 C ALAA 116 10.435 7.610 18.4001.00 10.70 6 ATOM 878 O ALAA 116 10.537 6.377 18.3971.00 11.22 8 ATOM 879 CB ALAA 116 8.292 8.902 18.1841.00 14.75 6 ATOM 880 N SERA 117 11.370 8.395 17.8481.00 12.15 7 ATOM 881 CA SERA 117 12,490 7.738 17.1501.00 12.96 6 ATOM 882 C SERA 117 13.387 6.913 18.0931.00 10.71 6 10ATOM 883 O SERA 117 13.805 5.814 17.7041.00 13.52 8 ATOM 884 CB SERA 117 13.345 8.825 16.5101.00 15.02 6 ATOM 885 OG SERA 117 12.600 9.369 15.4051.00 17.64 8 ATOM 886 N GLYA 118 13.537 7.392 19.3431.00 11.17 7 ATOM 887 CA GLYA 118 14.357 6.612 20.3011.00 11.65 6 --15ATOM 888 C GLYA 118 13.617 5.336 20.7071.00 12.79 6 ATOM 889 O GLYA 118 14.241 4.305 20.8871.00 11.77 8 ATOM 890 N ILEA 119 12.284 5.358 20.8691.00 10.37 7 ATOM 891 CA ILEA 119 11.517 4.140 21.1641.00 9.25 6 ATOM 892 C ILEA 119 11.754 3.131 20.0161.00 9.88 6 20ATOM 893 O ILEA 119 11.966 1.949 20.3171.00 9.88 8 ATOM 894 CB ILEA 119 10.045 4.484 21.3371.00 8.90 6 ATOM 895 CG1 ILEA 119 9.871 5.274 22.7141.00 10.96 6 ATOM 896 CG2 ILEA 119 9.131 3.264 21.3061.00 10.98 6 ATOM 897 CD1 ILEA 119 8.439 5.822 22.7681.00 11.18 6 25ATOM 898 N ARGA 120 11.557 3.597 18.7561.00 9.24 7 ATOM 899 CA ARGA 120 11.799 2.616 17.6831.00 10.97 6 ATOM 900 C ARGA 120 13.239 2.125 17.6521.00 9.58 6 ATOM 901 O ARGA 120 13.447 0.905 17.4821.00 11.11 8 ATOM 902 CB ARGA 120 11.497 3.354 16.3361.00 10.45 6 30ATOM 903 CG ARGA 120 10.021 3.739 16.2151,00 12.73 6 ATOM 904 CD ARGA 120 9.770 4.717 14.9881.00 12.77 6 ATOM 905 NE ARGA 120 9.$24 3.724 13.9111.00 13.28 7 ATOM 906 CZ ARGA 120 8.753 3.068 13.5121.00 11.79 6 ATOM 907 NH1 ARGA 120 7.523 3.292 13.8961.00 12.69 7 35ATOM 908 NH2 ARGA 120 8.965 2.095 12.6381.00 11.60 7 ATOM 909 N TYRA 121 14.187 3.005 17.9341.00 10.24 7 ATOM 910 CA TYRA 121 15.594 2.588 17.9881.00 12.90 6 ATOM 911 C TYRA 121 15.860 1.471 18.9691.00 9.83 6 ATOM 912 O TYRA 121 16.522 0.447 18.7881.00 11.54 8 40ATOM 913 CB TYRA 121 16.416 3.837 18.2921.00 12.06 6 ATOM 914 CG TYRA 121 17.853 3.571 18.5961.00 11.82 6 ATOM 915 CD1 TYRA 121 18.818 3.475 17.6041.00 13.12 6 ATOM 916 CD2 TYRA 121 18.273 3.395 19.8961.00 11.97 6 ATOM 917 CE1 TYRA 121 20.157 3.225 17.9301.00 13.85 6 45ATOM 918 CE2 TYRA 121 19.575 3.177 20.2501.00 11.79 6 ATOM 919 CZ TYRA 121 20.518 3.073 19.2521.00 15.08 6 ATOM 920 OH TYRA 121 21.856 2.849 19.5851.00 17.95 8 ATOM 921 N ALAA 122 15.231 1.676 20.1661.00 9.11 7 ATOM 922 CA ALAA 122 15.446 0.670 21.1971.00 9.16 6 50ATOM 923 C ALAA 122 14.894 -0.675 20.7741.00 10.24 6 ATOM 924 0 ALAA 122 15.500 -1.733 21.0451.00 12.08 8 ATOM 925 CB ALAA 122 14.726 1.101 22.4811.00 10.97 6 ATOM 926 N ALAA 123 13.672 -0.746 20.1601.00 10.00 7 ATOM 927 CA ALAA 123 13.177 -2.032 19.6601.00 9.79 6 55ATOM 928 C ALAA 123 14.082 -2.548 18.5221.00 10.77 6 ATOM 929 O ALAA 123 14.298 -3.791 18.4641.00 12.07 8 ATOM 930 CB ALAA 123 11.747 -1.789 19.1351.00 12.17 6 ATOM 931 N ASPA 124 14.513 -1.608 17.6841.00 10.69 7 ATOM 932 CA ASPA 124 15.338 -2.079 16.5481.00 11.25 6 60ATOM 933 C ASPA 124 16.699 -2.611 17.0141.00 11.36 6 ATOM 934 0 ASPA 124 17.263 -3.536 16.3721.00 12.55 8 ATOM 935 CB ASPA 124 15.528 -0.967 15.5271.00 10.62 6 ATOM 936 CG ASPA 124 14.197 -0.704 14.7271.00 11.72 6 ATOM 937 OD1 ASPA 124 13.461 -1.679 14.6091.00 13.58 8 65ATOM 938 OD2 ASPA 124 14.060 0.480 14.3521.00 13.00 8 ATOM 939 N GLNA 125 17.178 -2.140 18.1521.00 10.69 7 ATOM 940 CA GLNA 125 18.385 -2.681 18.7721.00 11.00 6 ATOM 941 C GLNA 125 18.156 -3.958 19.5271.00 10.13 6 ATOM 942 O GLN A 125 19.112 -4.519 20.1141.00 14.93 8 ATOM 943 CB GLN A 125 19.045 -1.632 19.6901.00 13.56 6 ATOM 944 CG GLN A 125 19.636 -0.472 18.9001.00 15.15 6 ATOM 945 CD GLN A 125 20.953 -0.735 18.1921.00 21.96 6 ATOM 946 OE1GLN A 125 21.571 -1.784 18.2911.00 28.15 8 ATOM 947 NE2GLN A 125 21.464 0.234 17.4331.00 29.01 7 ATOM 948 N GLY A 126 16.930 -4.457 19.6661.00 9.97 7 ATOM 949 CA GLY A 126 16.710 -5.768 20.2791.00 11.91 6 ATOM 950 C GLY A 126 16.402 -5.656 21.7891.00 9.69 6 10ATOM 951 O GLY A 126 16.461 -6.756 22.3471.00 11.42 8 ATOM 952 N ALA A 127 16.191 -4.473 22.3111.00 10.80 7 ATOM 953 CA ALA A 127 15.916 -4.514 23.7751.00 11.01 6 ATOM 954 C ALA A 127 14.656 -5.298 24.0941.00 11.87 6 ATOM 955 O ALA A 127 13.625 -5.169 23.3821.00 11.59 8 15ATOM 956 CB ALA A 127 15.812 -3.046 24.2411.00 11.61 6 ATOM 957 N LYS A 128 14.714 -6.115 25.1931.00 10.72 7 ATOM 958 CA LYS A 128 13.507 -6.851 25.5201.00 10.65 6 ATOM 959 C LYS A 128 12.450 -6.045 26.2741.00 8.78 6 ATOM 960 O LYS A 128 11.270 -6.377 26.2011.00 10.35 8 20ATOM 961 CB LYS A 128 13.834 -8.046 26.4421.00 12.01 6 ATOM 962 CG LYS A 128 14.845 -9.003 25.8411.00 17.41 6 ATOM 963 CD LYS A 128 14.181 -9.713 24.6631.00 17.61 6 ATOM 964 CE LYS A 128 15.182 -10.78124.1741.00 24.85 6 ATOM 965 NZ LYS A 128 14.810 -11.33322.8351.00 23.69 7 25ATOM 966 N VAL A 129 12.912 -4.970 26.9051.00 9.09 7 ATOM 967 CA VAL A 129 12.007 -4.094 27.6871.00 8.78 6 ATOM 968 C VAL A 129 12.468 -2.678 27.4061.00 8.25 6 ATOM 969 O VAL A 129 13.664 -2.390 27.3171.00 10.23 8 ATOM 970 CB VAL A 129 12.239 -4.362 29.1881.00 10.07 6 30ATOM 971 CG1VAL A 129 11.286 -3.527 30.0711.00 10.09 6 ATOM 972 CG2VAL A 129 11.977 -5.856 29.4681.00 10.08 6 ATOM 973 N LEU A 130 11.489 -1.779 27.2891.00 8.25 7 ATOM 974 CA LEU A 130 11.736 -0.350 27.1851.00 8.42 6 ATOM 975 C LEU A 130 11.104 0.353 28.4111.00 8.00 6 35ATOM 976 O LEU A 130 9.952 0.083 28.7841.00 9.41 B

ATOM 977 CB LEU A 130 11.008 0.264 25.9401.00 11.97 6 ATOM 978 CG LEU A 130 11.719 0.108 24.5791.00 10.92 6 ATOM 979 CD1LEU A 130 11.814 -1.346 24.1911.00 13.24 6 ATOM 980 CD2LEU A 130 10.890 0.862 23.5141.00 10.28 6 40ATOM 981 N ASN A 131 11.941 1.213 29.0651.00 7.78 7 ATOM 982 CA ASN A 131 11.363 1.989 30.1681.00 8.96 6 ATOM 983 C ASN A 131 11.240 3.469 29.7131.00 9.31 6 ATOM 984 O ASN A 131 12.244 4.033 29.2591.00 11.04 B

ATOM 985 CB ASN A 131 12.331 1.939 31.3721.00 9.40 6 45ATOM 986 CG ASN A 131 11.721 2.692 32.5371.00 9.88 6 ATOM 987 OD1ASN A 131 10.903 2.118 33.2691.00 10.13 8 ATOM 988 ND2ASN A 131 12.055 3.968 32.6611.00 9.63 7 ATOM 989 N LEU A 132 9.984 3.975 29.8801.00 8.49 7 ATOM 990 CA LEU A 13:.'9.726 5.379 29.5571.00 9.46 6 50ATOM 991 C LEU A 132 9.192 6.133 30.7881.00 9.52 6 ATOM 992 0 LEU A 13?. 8.007 6.230 31.0451.00 9.42 8 ATOM 993 CB LEU A 132 8.612 5.453 28.4661.00 9.79 6 ATOM 994 CG LEU A 132 9.154 4.944 27.0851.00 11.13 6 ATOM 995 CD1LEU A 132 8.014 4.418 26.2611.00 12.43 6 55ATOM 996 CD2LEU A 132 9.822 6.117 26.4081.00 15.07 6 ATOM 997 N SER A 133 10.203 6.676 31.5231.00 9.31 7 ATOM 998 CA SER A 133 9.908 7.485 32.7081.00 8.09 6 ATOM 999 C SER A 133 9.697 8.938 32.2191.00 9.51 6 ATOM 1000 O SER A 133 10.434 9.828 32.5701.00 12.68 8 60ATOM 1001 CB SER A 133 11.008 7.383 33.7521.00 10.34 6 ATOM 1002 OG SER A 133 10.943 6.119 34.4011.00 9.84 8 ATOM 1003 N LEU A 134 8.623 9.104 31.4291.00 9.63 7 ATOM 1004 CA LEU A 134 8.400 10.415 30.7621.00 9.62 6 ATOM 1005 C LEU A 134 6.942 10.351 30.2961.00 12.30 6 65ATOM 1006 0 LEU A 134 6.298 9.299 30.1471.00 11.68 8 ATOM 1007 CB LEU A 134 9.378 10.676 29.6121.00 12.20 6 ATOM 1008 CG LEU A 134 9.390 9.650 28.5001.00 11.16 6 ATOM 1009 CD1LEU A 134 8.275 9.976 27.4821.00 13.93 6 ATOM 1010 CD2 LEU A134 10.722 9.590 27.782 1.00 16.32 ATOM 1011 N GLY A135 6.429 11.54929.949 1.00 12.94 ATOM 1012 CA GLY A135 5.066 11.53129.372 1.00 16.25 ATOM 1013 C GLY A135 4.494 12.93729.388 1.00 20.76 ATOM 1014 O GLY A135 5.104 13.91129.837 1.00 19.61 ATOM 1015 N CYS A136 3.264 12.93428.855 1.00 16.71 ATOM 1016 CA CYS A136 2.541 14.22028.850 1.00 20.54 ATOM 1017 C CYS A136 1.077 13.96628.524 1.00 16.55 ATOM 1018 O CYS A136 0.649 12.88628.170 1.00 14.70 10ATOM 1019 CB CYS A136 3.085 15.19527.836 1.00 22.72 ATOM 1020 SG CYS A136 3.714 14.54626.303 1.00 26.03 ATOM 1021 N GLU A137 0.333 15.09328.682 1.00 18.30 ATOM 1022 CA GLU A137 -1.056 15.04028.175 1.00 17.88 ATOM 1023 C GLU A137 -1.003 15.55726.748 1.00 22.52 15ATOM 1024 O GLU A137 -1.289 16.70426.391 1.00 21.83 ATOM 1025 CB GLU A137 -2.021 15.83729.031 1.00 20.26 ATOM 1026 CG GLU A137 -2.281 15.29630.439 1.00 22.38 ATOM 1027 CD GLU A137 -3.418 16.13031.064 1.00 26.72 ATOM 1028 OE1 GLU A137 -3.051 17.08831.746 1.00 35.28 20ATOM 1029 OE2 GLU A137 -4.576 15.75730.819 1.00 21.07 ATOM 1030 N CYS A138 -0.616 14.67325.866 1.00 21.75 ATOM 1031 CA CYS A138 -0.209 14.96924.515 1.00 27.45 ATOM 1032 C CYS A138 -0.666 13.87323.581 1.00 31.92 ATOM 1033 O CYS A138 -0.656 12.70523.982 1.00 30.57 25ATOM 1034 CB CYS A138 1.332 15.10024.522 1.00 29.63 ATOM 1035 SG CYS A138 2.180 13.66425.316 1.00 26.98 ATOM 1036 N ASN A139 -1.166 14.25822.421 1.00 30.94 ATOM 1037 CA ASN A139 -1.597 13.30021.407 1.00 30.40 ATOM 1038 C ASN A139 -0.544 13.30820.306 1.00 27.83 30ATOM 1039 O ASN A139 -0.056 14.36219.882 1.00 27.73 B

ATOM 1040 CB ASN A139 -2.957 13.67620.845 1.00 34.15 ATOM 1041 CG ASN A139 -4.122 12.99321.530 1.00 44.83 ATOM 1042 OD1 ASN A139 -4.207 13.02522.756 1.00 38.38 ATOM 1043 ND2 ASN A139 -4.999 12.38920.735 1.00 50.90 35ATOM 1044 N SER A140 -0.166 12.12019.829 1.00 18.15 ATOM 1045 CA SER A140 0.870 12.08118.795 1.00 18.59 ATOM 1046 C SER A140 0.759 10.73818.087 1.00 16.05 ATOM 1047 O SER A140 0.941 9.697 18.736 1.00 16.64 ATOM 1048 CB SER A140 2.267 12.16519.402 1.00 20.72 40ATOM 1049 OG SER A140 3.309 11.90818.514 1.00 23.30 ATOM 1050 N THR A141 0.370 10.71216.804 1.00 18.63 ATOM 1051 CA THR A141 0.271 9.420 16.137 1.00 18.09 ATOM 1052 C THR A141 1.670 8.843 15.884 1.00 15.35 ATOM 1053 0 THR A141 1.750 7.578 15.886 1.00 15.18 45ATOM 1054 CB THR A141 -0.540 9.429 14.838 1.00 20.90 ATOM 1055 OGl THR A141 0.118 10.35813.966 1.00 23.57 ATOM 1056 CG2 THR A141 -1.990 9.800 15.085 1.00 17.57 ATOM 1057 N THR A142 2.720 9.626 15.861 1.00 16.63 ATOM 1058 CA THR A142 4.094 9.149 15.775 1.00 14.88 50ATOM 1059 C THR A142 4.529 8.429 17.053 1.00 13.11 ATOM 1060 O THR A142 5.094 7.345 16.979 1.00 14.05 ATOM 1061 CB THR A142 4.997 10.34115.457 1.00 24.16 ATOM 1062 OG1 THR A142 4.523 10.83514.153 1.00 28.29 ATOM 1063 CG2 THR A142 6.432 9.970 15.210 1.00 26.14 55ATOM 1064 N LEU A143 4.124 8.997 18.199 1.00 13.69 ATOM 1065 CA LEU A143 4.512 8.335 19.463 1.00 13.71 ATOM 1066 C LEU A143 3.729 7.043 19.617 1.00 12.67 ATOM 1067 O LEU A143 4.267 6.012 20.055 1.00 11.28 ATOM 1068 CB LEU A143 4.184 9.316 20.618 1.00 12.30 60ATOM 1069 CG LEU A143 4.738 8.845 21.995 1.00 15.95 ATOM 1070 CD1 LEU A143 6.227 8.878 22.029 1.00 15.41 ATOM 1071 CD2 LEU A143 4.099 9.884 22.948 1.00 15.72 ATOM 1072 N LYS A144 2.400 7.101 19.314 1.00 11.25 ATOM 1073 CA LYS A144 1.651 5.858 19.420 1.00 11.23 65ATOM 1074 C LYS A144 2.211 4.764 18.517 1.00 10.18 ATOM 1075 O LYS A144 2.312 3.600 18.899 1.00 10.89 ATOM 1076 CB LYS A144 0.159 6.178 19.140 1.00 15.25 ATOM 1077 CG LYS A144 -0.627 4.905 19.387 1.00 18.48 ATOM 1078 CD LYSA 144 -2.062 4.950 19.844 1.0024.85 ATOM 1079 CE LYSA 144 -2.564 3.597 20.366 1.0015.78 ATOM 1080 NZ LYSA 144 -2.599 2.616 19.228 1.0014.78 ATOM 1081 N SERA 145 2.539 5.151 17.273 1.0011.46 ATOM 1082 CA SERA 145 3.097 4.182 16.357 1.0010.88 ATOM 1083 C SERA 145 4.407 3.579 16.834 1.0010.64 ATOM 1084 0 SERA 145 4.628 2.364 16.771 1.0011.55 ATOM 1085 CB SERA 145 3.372 4.933 15.034 1.0011.53 ATOM 1086 OG SERA 145 4.095 4.059 14.159 1.0012.02 B

10ATOM 1087 N ALAA 146 5.223 4.406 17.500 1.0011.16 ATOM 1088 CA ALAA 146 6.521 3.907 17.961 1.0010.64 ATOM 1089 C ALAA 146 6.280 2.864 19.071 1.008.68 ATOM 1090 O ALAA 146 6.945 1.842 19.103 1.0010.24 ATOM 1091 CB ALAA 146 7.363 5,053 18.531 1.0013.58 15ATOM 1092 N VALA 147 5.345 3.181 19.973 1.009.25 ATOM 1093 CA VALA 147 5.063 2.216 21.057 1.0010.47 ATOM 1094 C VALA 147 4.479 0.936 20.530 1.009.13 ATOM 1095 0 VALA 147 4.842 -0.17220.914 1.0011.53 ATOM 1096 CB VALA 147 4.051 2.840 22.072 1.009.11 20ATOM 1097 CG1 VALA 147 3.468 1.828 23.057 1.0010.14 ATOM 1098 CG2 VALA 147 4.741 3.918 22.848 1.0011.38 ATOM 1099 N ASPA 148 3.531 1.044 19.539 1.0010.88 ATOM 1100 CA ASPA 148 2.945 -0.15818.998 1.009.20 ATOM 1101 C ASPA 148 3.904 -0.98618.148 1.0010.40 25ATOM 1102 O ASPA 148 3.989 -2.21618.235 1.0012.08 ATOM 1103 CB ASPA 148 1.722 0.191 18.150 1.009.51 ATOM 1104 CG ASPA 148 0.523 0.649 18.916 1.0012.65 ATOM 1105 OD1 ASPA 148 -0.363 1.347 18.361 1.0013.89 ATOM 1106 OD2 ASPA 148 0.454 0.337 20.139 1.0012.46 30ATOM 1107 N TYRA 149 4.776 -0.20317.443 1.0010.28 ATOM 1108 CA TYRA 149 5.839 -0.92016.701 1.0011.03 ATOM 1109 C TYRA 149 6.725 -1.73517.654 1.0011.91 ATOM 1110 0 TYRA 149 7.097 -2.87017.371 1.0011.38 ATOM 1111 CB TYRA 149 6.606 0.125 15.893 I.009.52 35ATOM 1112 CG TYRA 149 7.854 -0.42515.218 1.009.13 ATOM 1113 CD1 TYRA 149 7.714 -1.15614.034 1.0013.04 ATOM 1114 CD2 TYRA 149 9.133 -0.22015.669 1.009.76 ATOM 1115 CE1 TYRA 149 8.844 -1.65513.380 1.0011.71 ATOM 1116 CE2 TYRA 149 10.277 -0.69215.036 1.0010.97 40ATOM 1117 CZ TYRA 149 10.099 -1.41513.843 1.0013.29 ATOM 1118 OH TYRA 149 11.230 -1.87913.246 1.0012.70 ATOM 1119 N ALAA 150 7.149 -1.06118.759 1.0010.29 ATOM 1120 CA ALAA 150 8.086 -1.80119.644 1.0010.75 ATOM 1121 C ALAA 150 7.409 -3.01420.263 1.0012.48 45ATOM 1122 O ALAA 150 7.986 -4.10220.400 1.0011.16 ATOM 1123 CB ALAA 150 8.414 -0.86720.792 1.0011.39 ATOM 1124 N TRPA 151 6.140 -2.88220.642 1.009.91 ATOM 1125 CA TRPA 151 5.362 -4.01521.124 1.0010.13 ATOM 1126 C TRPA 151 5.261 -5.14620.085 1.009.48 50ATOM 1127 0 TRPA 151 5.539 -6.30520.358 1.0010.95 ATOM 1128 CB TRPA 151 3.927 -3.58221.579 1.0010.48 ATOM 1129 CG TRPA 151 3,161 -4.78521.971 1.0011.85 ATOM 1130 CD1 TRPA 151 2.277 -5.51121.197 1.0013.81 ATOM 1131 CD2 TRPA 151 3.146 -5.49023.235 1.009.96 55ATOM 1132 NE1 TRPA 151 1.771 -6.61221.846 1.0015.34 ATOM 1133 CE2 TRPA 151 2.285 -6.59223.111 1.0011.52 ATOM 1134 CE3 TRPA 151 3.799 -5.27024.452 1.0012.17 ATOM 1135 CZ2 TRPA 151 2.055 -7.48724.161 1.0011.76 ATOM 1136 CZ3 TRPA 151 3.603 -6.16125.520 1.0014.83 60ATOM 1137 CH2 TRPA 151 2.747 -7.23525.354 1.0011.91 ATOM 1138 N ASNA 152 4.921 -4.75818.850 1.0010.31 ATOM 1139 CA ASNA 152 4.758 -5.80517.805 1.0010.90 ATOM 1140 C ASNA 152 6.078 -6.36517.381 1.0012.51 ATOM 1141 O ASNA 152 6.094 -7.49816.850 1.0018.29 65ATOM 1142 CB ASNA 152 4.057 -5.13816.614 1.0012.21 ATOM 1143 CG ASNA 152 2.596 -4.89816.919 1.0015.60 ATOM 1144 OD1 ASNA 152 1.888 -5.69717.581 1.0016.16 ATOM 1145 ND2 ASNA 152 2.084 -3.80716.394 1.0016.47 ATOM 1146 N LYSA 153 7.214 -5.769 17.698 1.0012.60 ATOM 1147 CA LYSA 153 8.552 -6.282 17.497 1.0012.34 ATOM 1148 C LYSA 153 8.890 -7.341 18.558 1.0011.63 ATOM 1149 O LYSA 153 9.908 -8.014 18.454 1.0016.70 5 ATOM 1150 CB LYSA 153 9.587 -5.158 17.537 1.0014.85 ATOM 1151 CG LYSA 153 9.633 -4.265 16.316 1.0021.79 ATOM 1152 CD LYSA 153 10.522 -4.776 15.210 1.0020.60 ATOM 1153 CE LYSA 153 12.016 -4.864 15.642 1.0014.64 ATOM 1154 NZ LYSA 153 12.600 -5.708 14.521 1.0023.18 10ATOM 1155 N GLYA 154 8.101 -7.351 19.658 1.0011.47 ATOM 1156 CA GLYA 154 8.345 -8.288 20.722 1.0010.77 ATOM 1157 C GLYA 154 8.817 -7.664 22.030 1.0010.93 ATOM 1158 O GLYA 154 9.088 -8.468 22.922 1.0012.36 ATOM 1159 N ALAA 155 8.909 -6.356 22.134 1.0010.32 15ATOM 1160 CA ALAA 155 9.381 -5.744 23.380 1.009.51 ATOM 1161 C ALAA 155 8.226 -5.469 24.340 1.0010.33 ATOM 1162 O ALAA 155 7.074 -5.268 23.959 1.0010.45 ATOM 1163 CB ALAA 155 10.101 -4.418 22.994 1.0010.15 ATOM 1164 N VALA 156 8.529 -5.419 25.626 1.0010.20 20ATOM 1165 CA VALA 156 7.608 -4.930 26.644 1.009.64 ATOM 1166 C VALA 156 7.900 -3.424 26.826 1.0011.32 ATOM 1167 O VALA 156 9.077 -3.071 26.918 1.0011.90 ATOM 1168 CB VALA 156 7.867 -5.665 27.974 1.008.78 ATOM 1169 CG1 VALA 156 6.965 -5.108 29.060 1.008.99 25ATOM 1170 CG2 VALA 156 7.629 -7.139 27.784 1.0010.63 ATOM 1171 N VALA 157 6.818 -2.653 26.805 1.008.38 ATOM 1172 CA VALA 157 6.923 -1.208 27.010 1.007.61 ATOM 1173 C VALA 157 6.322 -0.872 28.402 1.007.95 ATOM 1174 O VALA 157 5.156 -1.202 28.638 1.009.65 30ATOM 1175 CB VALA 157 6.194 -0.421 25.888 1.009.67 ATOM 1176 CG1 VALA 157 6.257 1.064 26.099 1.0011.64 ATOM 1177 CG2 VALA 157 6.789 -0.831 24.528 1.0011.35 ATOM 1178 N VALA 158 7.116 -0.189 29.203 1.008.45 ATOM 1179 CA VALA 158 6.700 0.190 30.574 1.009.13 35ATOM 1180 C VALA 158 6.807 1.706 30.663 1.009.56 ATOM 1181 O VALA 158 7.873 2.222 30.248 1.009.44 ATOM 1182 CB VALA 158 7.639 -0.489 31.598 1.008.39 ATOM 1183 CG1 VALA 158 7.139 -0.079 33.007 1.009.17 ATOM 1184 CG2 VALA 158 7.635 -2.003 31.414 1.009.94 40ATOM 1185 N ALAA 159 5.799 2.385 31.165 1.008.36 ATOM 1186 CA ALAA 159 5.874 3.865 31.227 1.008.80 ATOM 1187 C ALAA 159 5.232 4.389 32.506 1.009.68 ATOM 1188 O ALAA 159 4.251 3.858 33.049 1.009.19 ATOM 1189 CB ALAA 159 5.122 4.442 30.023 1.0011.81 45ATOM 1190 N ALAA 160 5.842 5.486 32.979 1.0010.16 ATOM 1191 CA ALAA 160 5.305 6.213 34.150 1.008.09 ATOM 1192 C ALAA 160 3.970 6.867 33.890 1.0010.59 ATOM 1193 O ALAA 160 3.740 7.477 32.843 1.0012.25 ATOM 1194 CB ALAA 160 6.379 7.244 34.509 1.0010.91 50ATOM 1195 N ALAA 161 3.077 6.756 34.901 1.0010.18 ATOM 1196 CA ALAA 161 1.740 7.326 34.667 1.009.20 ATOM 1197 C ALAA 161 1.681 8.838 34.846 1.009.69 ATOM 1198 O ALAA 161 0.615 9.381 34.494 1.0011.99 B

ATOM 1199 CB ALAA 161 0.757 6.678 35.666 1.0011.17 55ATOM 1200 N GLYA 162 2.697 9.461 35.379 1.0010.16 ATOM 1201 CA GLYA 162 2.728 10.929 35.525 1.0011.47 ATOM 1202 C GLYA 162 2.542 11.334 36.997 1.0011.99 ATOM 1203 O GLYA 162 2.058 10.534 37.818 1.0011.61 ATOM 1204 N ASNA 163 2.830 12.646 37.210 1.0013.68 60ATOM 1205 CA ASNA 163 3.016 13.100 38.616 1.0014.00 ATOM 1206 C ASNA 163 2.233 14.384 38.911 1.0014.39 ATOM 1207 O ASNA 163 2.725 15.174 39.760 1.0018.79 ATOM 1208 CB ASNA 163 4.477 13.375 38.878 1.0017.01 ATOM 1209 CG ASNA 163 5.442 12.263 38.552 1.0020.97 65ATOM 1210 OD1 ASNA 163 5.223 11.120 38.907 1.0022.23 ATOM 1211 ND2 ASNA 163 6.522 12.569 37.843 1.0040.92 ATOM 1212 N ASPA 164 1.039 14.444 38.394 1.0014.22 ATOM 1213 CA ASPA 164 0.248 15.664 38.640 1.0015.43 ATOM 1214 C ASP A164 -0.891 15.376 39.6101.00 16.52 ATOM 1215 0 ASP A164 -1.808 16.205 39.7911.00 16.92 ATOM 1216 CB ASP A164 -0.340 16.092 37.3041.00 18.79 ATOM 1217 CG ASP A164 0.611 16.817 36.3821.00 31.26 ATOM 1218 OD1 ASP A164 0.099 17.435 35.4371.00 32.77 ATOM 1219 OD2 ASP A164 1.843 16.799 36.5781.00 32.99 ATOM 1220 N ASN A165 -0.956 14.222 40.2281.00 13.48 ATOM 1221 CA ASN A165 -2.032 13.774 41.0471.00 13.19 ATOM 1222 C ASN A165 -3.417 13.950 40.4241.00 12.14 10ATOM 1223 O ASN A165 -4.334 14.510 41.0361.00 15.12 ATOM 1224 CB ASN A165 -2.028 14.587 42.3691.00 12.06 ATOM 1225 CG ASN A165 -2.933 13.893 43.3481.00 10.24 ATOM 1226 OD1 ASN A165 -3.244 12.729 43.4791.00 12.45 ATOM 1227 ND2 ASN A165 -3.428 14.777 44.2971.00 11.65 15ATOM 1228 N VAL A166 -3.533 13.571 39.1691.00 12.64 ATOM 1229 CA VAL A166 -4.803 13.600 38.4421.00 13.30 ATOM 1230 C VAL A166 -5.190 12.205 37.9081.00 13.31 ATOM 1231 O VAL A165 -4.366 11.280 37.8551.00 11.72 ATOM 1232 CB VAL A166 -4.852 14.651 37.3301.00 15.75 20ATOM 1233 CG1 VAL A166 -4.413 16.035 37.8171.00 19.38 ATOM 1234 CG2 VAL A166 -3.879 14.327 36.2051.00 15.73 ATOM 1235 N SER A167 -6.430 12.097 37.4251.00 15.05 ATOM 1236 CA SER A167 -6.858 10.862 36.7531.00 14.74 ATOM 1237 C SER A167 -7.051 10.847 35.2661.00 11.97 25ATOM 1238 O SER A16? -7.439 9.759 34.8331.00 14.98 ATOM 1239 CB SER A167 -8.159 10.374 37.4531.00 21.93 ATOM 1240 OG SER A167 -9.169 11.371 37.2311.00 21.70 ATOM 1241 N ARG A168 -6.733 12.019 34.7601.00 14.80 ATOM 1242 CA ARG A168 -6.628 12.185 33.3361.00 14.30 30ATOM 1243 C ARG A168 -5.557 11.225 32.7611.00 14.04 ATOM 1244 O ARG A168 -4.583 11.016 33.4721.00 16.56 ATOM 1245 CB ARG A168 -6.450 13.600 32.8681.00 14.72 ATOM 1246 CG ARG A168 -7.590 14.492 33.4121.00 17.52 ATOM 1247 CD ARG A168 -7.488 15.866 32.7441.00 19.22 35ATOM 1248 NE ARG A168 -6.152 16.434 32.8541.00 19.87 ATOM 1249 CZ ARG A168 -5.777 17.137 33.9461.00 18.90 ATOM 1250 NH1 ARG A168 -6.683 17.234 34.9151.00 22.35 ATOM 1251 NH2 ARG A168 -4.590 17.669 34.0371.00 27.26 ATOM 1252 N THR A169 -5.775 10.681 31.5451.00 13.30 40ATOM 1253 CA THR A169 -4.663 9.851 31.0361.00 14.18 ATOM 1254 C THR A169 -3.476 10.689 30.6531.00 14.82 ATOM 1255 0 THR A169 -3.422 11.859 30.2201.00 16.21 ATOM 1256 CB THR A169 -5.168 9.132 29.7521.00 14.96 ATOM 1257 OG1 THR A169 -5.576 10.096 28.7541.00 15.86 45ATOM 1258 CG2 THR A169 -6.305 8.184 30.0461.00 17.51 ATOM 1259 N PHE A170 -2.290 10.036 30.7081.00 12.69 ATOM 1260 CA PHE A170 -0.978 10.559 30.3501.00 10.15 ATOM 1261 C PHE A170 -0.366 9.550 29.3821.00 10.51 ATOM 1262 O PHE A170 -0.516 8.340 29.5171.00 13.04 50ATOM 1263 CB APHEA170 0.010 10.860 31.4860.50 9.69 ATOM 1264 CG APHEA170 -0.086 12.208 32.1510.50 12.91 ATOM 1265 CD1 APHEA170 1.046 12.996 32.2470.50 15.42 ATOM 1266 CD2 APHEA170 -1.271 12.657 32.7230.50 15.96 ATOM 1267 CE1APHE A170 0.999 14.230 32.8930.50 17.60 55ATOM 1268 CE2APHE A170 -1.322 13.894 33.3590.50 14.69 ATOM 1269 CZ APHEA170 -0.193 14.664 33.4340.50 18.78 ATOM 1270 CB BPHEA170 -0.239 10.454 31.7130.50 11.27 ATOM 1271 CG BPHEA170 1.070 11.133 31.8300.50 10.40 ATOM 1272 CD1 BPHEA170 2.277 10.418 31.8530.50 10.22 60ATOM 1273 CD2 BPHEA170 1.133 12.520 31.9390.50 13.93 ATOM 1274 CE1 BPHEA170 3.482 11.075 31.9680.50 12.32 ATOM 1275 CE2 BPHEA170 2.348 13.165 32.0520.50 13.83 ATOM 1276 CZ BPHEA170 3.544 12.456 32.0770.50 15.52 ATOM 1277 N GLN A171 0.238 10.113 28.3311.00 11.14 65ATOM 1278 CA GLN A171 0.856 9.335 27.2551.00 10.59 ATOM 1279 C GLN A171 2.348 9.422 27.2391.00 10.58 ATOM 1280 O GLN A171 2.822 10.459 27.6451.00 13.39 ATOM 1281 CB GLN A171 0.297 9.849 25.8651.00 11.19 ATOM 1282 CG GLNA 171 -1.200 9.613 25.647 1.00 11.83 6 ATOM 1283 CD GLNA 171 -2.121 10.46826.524 1.00 13.12 6 ATOM 1284 OE1 GLNA 177. -2.934 9.928 27.305 1.00 15.76 8 ATOM 1285 NE2 GLNA 177. -2.011 11.79026.391 1.00 14.42 7 ATOM 1286 N PROA 172 3.043 8.320 26.919 1.00 11.03 7 ATOM 1287 CA PROA 172 2.572 7.108 26.347 1.00 11.46 6 ATOM 1288 C PROA 172 2.006 6.025 27.235 1.00 11.03 6 ATOM 1289 O PROA 172 1.509 5.023 26.809 1.00 11.22 8 ATOM 1290 CB PROA 172 3.819 6.511 25.610 1.00 11.89 6 10ATOM 1291 CG PROA 172 4.908 6.978 26.569 1.00 11.81 6 ATOM 1292 CD PROA 172 4.490 8.404 26.935 1.00 12.12 6 ATOM 1293 N ALAA 173 2.069 6.254 28.594 1.00 9.12 7 ATOM 1294 CA ALAA 173 1.563 5.194 29.473 1.00 9.14 6 ATOM 1295 C ALAA 173 0.120 4.778 29.127 1.00 10.86 6 w 15ATOM 1296 0 ALAA 173 -0.157 3.582 29.296 1.00 11.28 8 ATOM 1297 CB ALAA 173 1.640 5.713 30.931 1.00 11.79 6 ATOM 1298 N SERA 174 -0.804 5.696 28.751 1.00 9.25 7 ATOM 1299 CA SERA 174 -2.184 5.257 28.562 1.00 10.77 6 ATOM 1300 C SERA 174 -2.434 4.456 27.262 1.00 11.85 6 20ATOM 1301 O SERA 174 -3.559 3.938 27.120 1.00 12.90 8 ATOM 1302 CB SERA 174 -3.071 6.504 28.585 1.00 12.82 6 ATOM 1303 OG SERA 174 -2.887 7.251 27.376 1.00 13.72 8 ATOM 1304 N TYRA 175 -1.417 4.375 26.405 1.00 11.48 7 ATOM 1305 CA TYRA 175 -1.695 3.533 25.228 1.00 12.72 6 25ATOM 1306 C TYRA 175 -1.927 2.094 25.655 1.00 10.63 6 ATOM 1307 O TYRA 175 -1.259 1.611 26.603 1.00 11.10 8 ATOM 1308 CB TYRA 175 -0.435 3.567 24.316 1.00 11.23 6 ATOM 1309 CG TYRA 175 -0.129 4.914 23.750 1.00 10.17 6 ATOM 1310 CD1 TYRA 175 -1.068 5.887 23.517 1.00 13.17 6 30ATOM 1311 CD2 TYRA 175 1.198 5.229 23.425 1.00 9.64 6 ATOM 1312 CE1 TYRA 175 -0.763 7.116 22.977 1.00 11.74 6 ATOM 1313 CE2 TYRA 175 1.529 6.450 22.873 1.00 11.86 6 ATOM 1314 CZ TYRA 175 0.574 7.397 22.648 1.00 15.00 6 ATOM 1315 OH TYRA 175 0.880 8.636 22.122 1.00 16.46 8 35ATOM 1316 N PROA 176 -2.776 1.327 25.028 1.00 11.70 7 ATOM 1317 CA PROA 176 -2.959 -0.09225.360 1.00 11.01 6 ATOM 1318 C PROA 176 -1.660 -0.87825.46A 1.00 10.13 6 ATOM 1319 O PROA 176 -1.615 -1.76626.289 1.00 11.27 8 ATOM 1320 CB PROA 176 -3.990 -0.66524.357 1.00 12.68 6 40ATOM 1321 CG PROA 176 -4.746 0.637 24.093 1.00 11.38 6 ATOM 1322 CD PROA 176 -3.780 1.833 24.052 1.00 13.11 6 ATOM 1323 N ASNA 177 -0.723 -0.65624.506 1.00 11.05 7 ATOM 1324 CA ASNA 177 0.441 -1.54424.522 1.00 12.20 6 ATOM 1325 C ASNA 177 1.551 -1.04525.410 1.00 11.24 6 45ATOM 1326 O ASNA 177 2.629 -1.72025.402 1.00 11.19 8 ATOM 1327 CB ASNA 177 0.857 -1.64023.046 1.00 10.48 6 ATOM 1328 CG ASNA 177 0.051 -2.68422.321 1.00 12.91 6 ATOM 1329 OD1 ASNA 177 -0.414 -3.68922.832 1.00 14.78 8 ATOM 1330 ND2 ASNA 177 -0.019 -2.44120.970 1.00 15.54 7 50ATOM 1331 N ALAA 178 1.283 -0.05826.278 1.00 10.52 7 ATOM 1332 CA ALAA 178 2.264 0.293 27.312 1.00 9.20 6 ATOM 1333 C ALAA 178 1.728 -0.19128.662 1.00 11.35 6 ATOM 1334 O ALAA 178 0.548 0.004 28.907 1.00 10.58 8 ATOM 1335 CB ALAA 178 2.471 1.825 27.370 1.00 11.60 6 55ATOM 1336 N ILEA 179 2.559 -0.77929.554 1.00 9.76 7 ATOM 1337 CA ILEA 179 2.080 -0.94430.966 1.00 8.57 6 ATOM 1338 C ILEA 179 2.217 0.427 31.641 1.00 9.10 6 ATOM 1339 O ILEA 179 3.315 0.926 31.750 1.00 10.06 8 ATOM 1340 CB ILEA 179 3.011 -1.96131.637 1.00 8.69 6 60ATOM 1341 CG1 ILEA 179 2.926 -3.29230.879 1.00 11.26 6 ATOM 1342 CG2 ILEA 179 2.632 -2.22633.097 1.00 10.57 6 ATOM 1343 CD1 ILEA 179 3.905 -4.32331.403 1.00 13.43 6 ATOM 1344 N ALAA 180 1.097 0.950 32.181 1.00 8.93 7 ATOM 1345 CA ALAA 180 1.104 2.243 32.870 1.00 8.88 6 65ATOM 1346 C ALAA 180 1.312 1.988 34.378 1.00 9.81 6 ATOM 1347 O ALAA 180 0.623 1.134 34.956 1.00 10.08 8 ATOM 1348 CH ALAA 180 -0.257 2.936 32.657 1.00 10.99 6 ATOM 1349 N VALA 181 2.333 2.692 34.886 1.00 8.81 7 ATOM 1350 CA VALA 181 2.750 2.473 36.298 1.00 7.21 6 ATOM 1351 C VALA 181 2.625 3.698 37.175 1.00 10.43 6 ATOM 1352 O VALA 181 3.187 4.746 36.896 1.00 9.18 8 ATOM 1353 CB VALA 181 4.252 2.124 36.222 1.00 7.41 6 ATOM 1354 CG1 VALA 181 4.729 1.806 37.634 1.00 9.89 6 ATOM 1355 CG2 VALA 181 4.527 0.886 35.362 1.00 8.70 6 ATOM 1356 N GLYA 182 1.839 3.465 38.248 1.00 10.46 7 ATOM 1357 CA GLYA 182 1.639 4.475 39.285 1.00 9.36 6 ATOM 1358 C GLYA 182 2.682 4.200 40.403 1.00 10.52 6 10ATOM 1359 O GLYA 182 3.453 3.263 40.320 1.00 10.35 8 ATOM 1360 N ALAA 183 2.714 5.147 41.349 1.00 9.62 7 ATOM 1361 CA ALAA 183 3.677 4.975 42.430 1.00 8.49 6 ATOM 1362 C ALAA 183 2.990 4.939 43.792 1.00 10.18 6 ATOM 1363 O ALAA 183 2.028 5.635 44.041 1.00 10.96 8 w 15ATOM 1364 CB ALAA 183 4.536 6.262 42.471 1.00 11.75 6 ATOM 1365 N ILEA 184 3.671 4.126 44.619 1.00 8.08 7 ATOM 1366 CA ILEA 184 3.277 4.029 46.044 1.00 9.34 6 ATOM 1367 C ILEA 184 4.522 4.315 46.889 1.00 10.84 6 ATOM 1368 O ILEA 184 5.660 4.279 46.425 1.00 10.59 8 20ATOM 1369 CB ILEA 184 2.?65 2.623 46.440 1.00 9.26 6 ATOM 1370 CG1 ILEA 184 3.623 1.53? 45.777 1.00 9.29 6 ATOM 1371 CG2 ILEA 184 1.298 2.458 46.049 1.00 10.38 6 ATOM 1372 CD1 ILEA 184 3.337 0.145 46.343 1.00 9.89 6 ATOM 1373 N ASPA 185 4.246 4.604 48.177 1.00 9.31 7 25ATOM 1374 CA ASPA 185 5.388 4.755 49.122 1.00 11.76 6 ATOM 1375 C ASPA 185 5.646 3.419 49.776 1.00 9.50 6 ATOM 1376 O ASPA 185 5.128 2.363 49.400 1.00 11.06 8 ATOM 1377 CB ASPA 185 4.996 5.878 50.077 1.00 12.42 6 ATOM 1378 CG ASPA 185 3.878 5.520 51.008 1.00 16.62 6 30ATOM 1379 OD1 ASPA 185 3.498 4.359 51.188 1.00 19.16 8 ATOM 1380 OD2 ASPA 185 3.331 6.525 51.584 1.00 21.71 8 ATOM 1381 N SERA 186 6.557 3.525 50.791 1.00 10.02 7 ATOM 1382 CA SERA 186 6.943 2.275 51.483 1.00 11.07 6 ATOM 1383 C SERA 186 5.904 1.628 52.388 1.00 11.68 6 35ATOM 1384 O SERA 186 6.089 0.458 52.791 1.00 12.40 8 ATOM 1385 CB SERA 186 8.278 2.475 52.195 1.00 12.70 6 ATOM 1386 OG SERA 186 8.020 3.342 53.353 1.00 13.04 8 ATOM 1387 N ASNA 187 4.805 2.372 52.571 1.00 12.58 7 ATOM 1388 CA ASNA 187 3.674 1.829 53.284 1.00 11.75 6 40ATOM 1389 C ASNA 187 2.521 1.414 52.403 1.00 14.42 6 ATOM 1390 O ASNA 187 1.387 1.286 52.844 1.00 14.41 8 ATOM 1391 CB ASNA 187 3.208 2.885 54.299 1.00 13.43 6 ATOM 1392 CG ASNA 187 2.435 2.266 55.455 1.00 24.32 6 ATOM 1393 OD1 ASNA 187 2.664 1.109 55.787 1.00 28.21 8 45ATOM 1394 ND2 ASNA 187 1.561 3.083 56.015 1.00 24.99 7 ATOM 1395 N ASPA 188 2.816 1.287 51.095 1.00 12.93 7 ATOM 1396 CA ASPA 188 1.790 0.919 50.135 1.00 12.49 6 ATOM 1397 C ASPA 188 0.681 1.950 49.920 1.00 12.89 6 ATOM 1398 O ASPA 188 -0.362 1.549 49.382 1.00 16.50 8 50ATOM 1399 CB ASPA 188 1.210 -0.47850.410 1.00 14.69 6 ATOM 1400 CG ASPA 188 2.107 -1.66150.168 1.00 14.67 6 ATOM 1401 OD1 ASPA 188 3.257 -1.50349.644 1.00 15.16 8 ATOM 1402 OD2 ASPA 188 1.754 -2.82150.535 1.00 17.10 8 ATOM 1403 N ARGA 189 0.944 3.168 50.317 1.00 11.91 7 55ATOM 1404 CA ARGA 189 -0.057 4.193 50.068 1.00 11.81 6 ATOM 1405 C ARGA 189 0.318 4.940 48.809 1.00 10.44 6 ATOM 1406 O ARGA 189 1.490 5.032 48.450 1.00 11.22 8 ATOM 1407 CB ARGA 189 -0.070 5.188 51.257 1.00 12.95 6 ATOM 1408 CG ARGA 189 -0.635 4.385 52.458 1.00 19.11 6 60ATOM 1409 CD ARGA 189 -0.942 5.273 53.602 0.00 20.00 6 ATOM 1410 NE ARGA 189 -1.563 4.465 54.658 0.00 20.00 7 ATOM 1411 CZ ARGA 189 -2.073 5.120 55.718 0.00 20.00 6 ATOM 1412 NH1 ARGA 189 -2.009 6.439 55.778 0.00 20.00 7 ATOM 1413 NH2 ARGA 189 -2.641 4.429 56.712 0.00 20.00 7 65ATOM 1414 N LYSA 190 -0.725 5.371 48.044 1.00 12.65 7 ATOM 1415 CA LYSA 190 -0.437 6.168 46.830 1.00 13.29 6 ATOM 1416 C LYSA 190 0.475 7.292 47.111 1.00 12.33 6 ATOM 1417 O LYSA 190 0.366 8.054 48.112 1.00 12.55 8 ATOM 1418 CB LYSA 190 -1.739 6.688 46.193 1.0014.04 6 ATOM 1419 CG LYSA 190 -1.575 7.374 44.863 1.0013.43 6 ATOM 1420 CD LYSA 190 -2.892 8.042 44.365 1.0014.23 6 ATOM 1421 CE LYSA 190 -2.848 9.547 44.467 1.0011.74 6 ATOM 1422 NZ LYSA 190 -1.794 10.509 44.344 1.0014.74 7 ATOM 1423 N ALAA 191 1.539 7.488 46.284 1.009.54 7 ATOM 1424 CA ALAA 191 2.402 8.643 46.397 1.0010.14 6 ATOM 1425 C ALAA 191 1.569 9.962 46.232 1.0012.32 6 ATOM 1426 0 ALAA 191 0.650 9.922 45.406 1.0012.53 8 ATOM 1427 CB ALAA 191 3.479 8.638 45.324 1.0012.15 6 ATOM 1428 N SERA 192 1.965 10.976 46.997 1.0013.24 7 ATOM 1429 CA SERA 192 1.044 12.139 46.993 1.0015.23 6 ATOM 1430 C SERA 192 0.868 12.637 45.580 1.0011.65 6 ATOM 1431 0 SERA 192 -0.259 13.044 45.208 1.0011.90 B

ATOM 1432 CB SERA 192 1.586 13.158 48.008 1.0019.44 6 ATOM 1433 OG SERA 192 2.765 13.652 47.508 1.0023.70 8 ATOM 1434 N PHEA 193 1.863 12.658 44.721 1.0012.20 7 ATOM 1435 CA PHEA 193 1.810 13.199 43.381 1.0014.48 6 ATOM 1436 C PHEA 193 1.385 12.209 42.289 1.0013.11 6 ATOM 1437 O PHEA 193 1.238 12.595 .41.1351.0011.70 8 ATOM 1438 CB PHEA 193 3.310 13.582 43.106 1.0017.86 6 ATOM 1439 CG PHEA 193 4.249 12.384 43.353 1.0020.29 6 ATOM 1440 CD1 PHEA 193 4.287 11.322 42.438 1.0022.95 6 ATOM 1441 CD2 PHEA 193 5.040 12.214 44.448 1.008.24 6 ATOM 1442 CE1 PHEA 193 5.098 10.236 42.710 1.0019.68 6 ATOM 1443 CE2 PHEA 193 5.864 11.222 44.781 1.0021.76 6 ATOM 1444 CZ PHEA 193 5.910 10.164 43.860 1.0019.49 6 ATOM 1445 N SERA 194 1.240 10.942 42.667 1.0010.08 7 ATOM 1446 CA SERA 194 1.056 10.001 41.524 1.009.55 6 ATOM 1447 C SERA 194 -0.269 10.206 40.817 1.0010.73 6 ATOM 1448 0 SERA 194 -1.304 10.320 41.445 1.0011.77 8 ATOM 1449 CB SERA 194 1.096 8.580 42.077 1.0010.02 6 ATOM 1450 OG SERA 194 0.951 7.609 41.021 1.0011.46 8 ATOM 1451 N ASNA 195 -0.250 10.146 39.487 1.009.56 7 ATOM 1452 CA ASNA 195 -1.500 10.042 38.765 1.009.33 6 ATOM 1453 C ASNA 195 -2.095 8.658 39.068 1.0012.48 6 ATOM 1454 0 ASNA 195 -1.471 7.723 3'9.5991.0011.95 B

ATOM 1455 CB ASNA 195 -1.288 10.176 37.252 1.009.08 6 ATOM 1456 CG ASNA 195 -0.941 11.572 36.865 1.0011.88 6 ATOM 1457 OD1 ASNA 195 -1.104 12.515 37.608 1.0012.04 8 ATOM 1458 ND2 ASNA 195 -0.437 11.729 35.635 1.0012.30 7 ATOM 1459 N TYRA 196 -3.396 8.535 38.769 1.0011.14 7 ATOM 1460 CA TYRA 196 -4.117 7.344 39.186 1.009.82 6 ATOM 1461 C TYRA 196 -5.386 7.196 38.350 1.0013.03 6 ATOM 1462 O TYRA 196 -5.716 8.132 37.629 1.0014.68 8 ATOM 1463 CB TYRA 196 -4.544 7.490 40.681 1.0011.85 6 ATOM 1464 CG TYRA 196 -5.414 8.701 40.933 1.0012.37 6 ATOM 1465 CD1 TYRA 196 -4.871 9.945 41.165 1.0012.29 6 ATOM 1466 CD2 TYRA 196 -6.802 8.592 40.906 1.0011.59 6 ATOM 1467 CE1 TYRA 196 -5.612 11.084 41.371 1.0013.98 6 ATOM 1468 CE2 TYRA 196 -7.586 9.704 41.112 1.0015.81 6 ATOM 1469 CZ TYRA 196 -7.009 10.918 41.336 1.0015.20 6 ATOM 1470 OH TYRA 196 -7.817 12.040 41.542 1.0019.65 8 ATOM 1471 N GLYA 197 -5.882 5.993 38.403 1.0011.13 7 ATOM 1472 CA GLYA 197 -7.170 5.755 37.680 1.0011.68 6 ATOM 1473 C GLYA 197 -7.164 4.338 37.161 1.0012.24 6 ATOM 1474 O GLYA 197 -6.200 3.578 37.267 1.0013.15 8 ATOM 1475 N THRA 198 -8.311 3.905 36.542 1.0012.40 7 ATOM 1476 CA THRA 198 -8.425 2.531 36.111 1.0011.46 6 ATOM 1477 C THRA 198 -7.578 2.248 34.855 1.0012.92 6 ATOM 1478 O THRA 198 -7.329 1.063 34.613 1.0014.81 8 ATOM 1479 CB THRA 198 -9.918 2.151 35.818 1.0013.71 6 ATOM 1480 OG1 THRA 198 -10.361 2.988 34.785 1.0020.98 8 ATOM 1481 CG2 THRA 198 -10.785 2.300 37.056 1.0017.57 6 ATOM 1482 N TRPA 199 -7.107 3.325 34.195 1.0012.12 7 ATOM 1483 CA TRPA 199 -6.192 3.157 33.081 1.0012.81 6 ATOM 1484 C TRPA 199 -4.774 2.869 33.525 1.0011.15 6 ATOM 1485 0 TRPA 199 -3.896 2.557 32.737 1.0013.41 8 WO 00/22103 .PCT/DK99/00542 ATOM 1486 CB TRPA 199 -6.194 4.421 32.173 1.0011.96 ATOM 1487 CG TRPA 199 -5.?44 5.662 32.884 1.0011.22 ATOM 1488 CD1 TRPA 199 -6.470 6.564 33.633 1.0016.14 ATOM 1489 CD2 TRPA 199 -4.419 6.188 32.931 1.0010.56 5 ATOM 1490 NE1 TRPA 199 -5.702 7.565 34.144 1.0016.05 ATOM 1491 CE2 TRPA 199 -4.397 7.379 33.705 1.0014.41 ATOM 1492 CE3 TRPA 199 -3.218 5.745 32.325 1.0012.64 ATOM 1493 CZ2 TRPA 199 -3.239 8.128 33.914 1.0017.36 ATOM 1494 CZ3 TRPA 199 -2.092 6.505 32.566 1.0015.03 10ATOM 1495 CH2 TRPA 199 -2.081 7.654 33.347 1.0014.87 ATOM 1496 N VALA 200 -4.468 3.143 34.811 1.0010.34 ATOM 1497 CA VALA 200 -3.115 2.805 35.303 1.0012.70 ATOM 1498 C VALA 200 -3.087 1.330 35.623 1.0011.86 ATOM 1499 O VALA 200 -4.069 0.763 36.157 1.0014.02 B

15ATOM 1500 CB VALA 200 -2.815 3.607 36.568 1.0010.90 ATOM 1501 CG1 VALA 200 -1.476 3.218 37.118 1.0011.75 ATOM 1502 CG2 VALA 200 -2.907 5.097 36.300 1.0010.81 ATOM 1503 N ASPA 201 -2.127 0.525 35.152 1.009.45 ATOM 1504 CA ASPA 201 -2.205 -0.91735.269 1.0010.79 20ATOM 1505 C ASPA 201 -1.714 -1.42836.611 1.0011.73 ATOM 1506 0 ASPA 201 -2.428 -2.19337.259 1.0010.74 ATOM 1507 CB ASPA 201 -1.361 -1.53234.104 1.0010.60 ATOM 1508 CG ASPA 201 -2.014 -1.23732.785 1.0014.68 ATOM 1509 OD1 ASPA 201 -3.197 -1.60332.591 1.0012.39 25ATOM 1514 OD2 ASPA 201 -1.329 -0.64231.929 1.0011.24 B

ATOM 1511 N VALA 202 -0.478 -1.01636.955 1.0010.17 ATOM 1512 CA VALA 202 0.069 -1.49838.207 1.0010.76 ATOM 1513 C VALA 202 0.716 -0.33738.925 1.009.89 ATOM 1514 O VALA 202 0.872 0.733 38.352 1.009.72 30ATOM 1515 CB VALA 202 1.128 -2.59138.016 1.009.97 ATOM 1516 CG1 VALA 202 0.504 -3.84737.440 1.0012.13 ATOM 1517 CG2 VALA 202 2.283 -2.10437.130 1.0014.06 ATOM 1518 N THRA 203 1.041 -0.55240.192 1.009.69 ATOM 1519 CA THRA 203 1.817 0.426 40.984 1.008.33 35ATOM 1520 C THRA 203 3.076 -0.20841.542 1.008.20 ATOM 1521 O THRA 203 3.152 -1.45541.659 1.008.89 ATOM 1522 CB THRA 203 0.899 1.018 42.077 1.0010.11 ATOM 1523 OG1 THRA 203 1.528 2.183 42.573 1.009.97 ATOM 1524 CG2 THRA 203 0.604 0.003 43.200 1.0010.55 40ATOM 1525 N ALAA 204 4.060 0.625 41.900 1.008.43 ATOM 1526 CA ALAA 204 5.322 0.114 42.440 1.007.80 ATOM 1527 C ALAA 204 5.934 1.201 43.307 1.007.92 ATOM 1528 0 ALAA 204 5.639 2.403 43.176 1.008.87 ATOM 1529 CB ALAA 204 6.215 -0.21841.206 1.009.02 45ATOM 1530 N PROA 205 6.889 0.857 44.167 1.009.20 ATOM 1531 CA PROA 205 7.626 1.788 44.988 1.0010.66 ATOM 1532 C PROA 205 8.167 2.923 44.122 1.0010.80 ATOM 1533 O PROA 205 8.865 2.755 43.103 1.0010.53 ATOM 1534 CB PROA 205 8.810 0.934 45.486 1.0011.87 50ATOM 1535 CG PROA 205 8.066 -0.38645.743 1.009.61 ATOM 1536 CD PROA 205 7.222 -0.56844.424 1.0010.97 ATOM 1537 N GLYA 206 7.855 4.139 44.552 1.009.51 ATOM 1538 CA GLYA 206 8.265 5.328 43.848 1.008.77 ATOM 1539 C GLYA 206 8.571 6.543 44.702 1.009.82 55ATOM 1540 O GLYA 206 8.762 7.640 44.198 1.0012.47 ATOM 1541 N VALA 207 8.608 6.363 46.053 1.009.75 ATOM 1542 CA VALA 207 8.789 7.498 46.954 1.0011.01 ATOM 1543 C VALA 207 10.120 7.353 47.646 1.0010.23 ATOM 1544 O VALA 207 10.366 6.348 48.257 1.0011.51 60ATOM 1545 CB VALA 207 7.679 7.624 48.011 1.009.92 ATOM 1546 CG1 VALA 207 7.938 8.802 48.933 1.0010.74 ATOM 1547 CG2 VALA 207 6.369 7.832 47.256 1.0012.64 ATOM 1548 N ASNA 208 10.960 8.360 47.573 1.009.70 ATOM 1549 CA ASNA 208 12.257 8.31? 48.278 1.009.99 65ATOM 1550 C ASNA 208 13.030 7.058 47.981 1.009.86 ATOM 1551 0 ASNA 208 13.447 6.265 48.772 1.0010.57 ATOM 1552 CB ASNA 208 12.033 8.481 49.791 1.0012.47 ATOM 1553 CG ASNA 208 11.614 9.893 50.142 1.0016.00 ATOM 1554 OD1ASN A208 11.947 10.841 49.4871.00 17.81 8 ATOM 1555 ND2ASN A208 10.820 9.952 51.2251.00 23.41 7 ATOM 1556 N ILE A209 13.185 6.904 46.6481.00 11.72 7 ATOM 1557 CA ILE A209 13.934 5.767 46.0911.00 11.01 6 ATOM 1558 C ILE A209 15.425 6.097 45.9621.00 10.83 6 ATOM 1559 0 ILE A209 15.707 7.084 45.2531.00 10.52 8 ATOM 1560 CB ILE A209 13.406 5.365 44.7111.00 9.48 6 ATOM 1561 CG1ILE A209 11.918 4.982 44.8061.00 9.44 6 ATOM 1562 CG2ILE A209 14.242 4.273 44.0341.00 10.15 6 10ATOM 1563 CD1ILE A209 11.642 3.820 45.7621.00 9.01 6 ATOM 1564 N ALA A210 16.292 5.399 46.7051.00 8.36 7 ATOM 1565 CA ALA A210 17.741 5.665 46.4911.00 9.20 6 ATOM 1566 C ALA A210 18.266 5.060 45.1911.00 11.06 6 ATOM 1567 O ALA A210 18.035 3.879 45.0161.00 11.02 8 15ATOM 1568 CB ALA A210 18.464 5.038 47.6781.00 10.57 6 ATOM 1569 N SER A211 19.037 5.807 44.4081.00 10.47 7 ATOM 1570 CA SER A211 19.647 5.229 43.2091.00 9.26 6 ATOM 1571 C SER A211 20.849 6.075 42.8391.00 10.82 6 ATOM 1572 O SER A211 21.241 7.011 43.5731.00 12.70 8 20ATOM 1573 CB SER A211 18.572 5.218 42.0761.00 11.65 6 ATOM 1574 OG SER A211 19.186 4.429 41.0311.00 9.57 8 ATOM 1575 N THR A212 21.521 5.6$4 41.7761.00 9.45 7 ATOM 1576 CA THR A212 22.650 6.435 41.2371.00 9.57 6 ATOM 1577 C THR A212 22.316 7.786 40.6011.00 10.33 6 25ATOM 1578 0 THR A212 21.312 7.897 39.9431.00 11.17 8 ATOM 1579 CB THR A212 23.230 5.539 40.1231.00 10.82 6 ATOM 1580 OG1THR A212 22.197 5.032 39.2551.00 11.01 B

ATOM 1581 CG2THR A212 23.860 4.247 40.6831.00 11.54 6 ATOM 1582 N VAL A213 23.202 8.751 40.8541.00 11.50 7 30ATOM 1583 CA VAL A213 23.084 10.032 40.1121.00 10.64 6 ATOM 1584 C VAL A213 24.479 10.365 39.6181.00 12.70 6 ATOM 1585 0 VAL A213 25.449 9.733 40.0231.00 14.01 B

ATOM 1586 CB VAL A213 22.430 11.117 40.9291.00 10.58 6 ATOM 1587 CG1VAL A213 20.952 10.866 41.2361.00 15.95 6 35ATOM 1588 CG2VAL A213 23.176 11.444 42.2121.00 16.49 6 ATOM 15$9 N PRO A214 24.599 11.271 38.6341.00 12.29 7 ATOM 1590 CA PRO A214 25.893 11.455 38.0321.00 14.14 6 ATOM 1591 C PRO A214 27.016 11.909 38.9591.00 15.71 6 ATOM 1592 O PRO A214 26.744 12.410 40.0521.00 16.38 8 40ATOM 1593 CB PRO A214 25.641 12.502 36.9191.00 15.16 6 ATOM 1594 CG PRO A214 24.175 12.119 36.5851.00 11.84 6 ATOM 1595 CD PRO A214 23.489 11.956 37.9691.00 14.36 6 ATOM 1596 N ASN A215 28.217 11.674 38.4511.00 17.19 7 ATOM 1597 CA ASN A215 29.421 12.081 39.2281.00 19.26 6 45ATOM 1598 C ASN A215 29.493 11.275 40.5141.00 18.85 6 ATOM 1599 0 ASN A215 29.781 11.814 41.5951.00 21.66 8 ATOM 1600 CB ASN A218 29.417 13.592 39.4931.00 19.94 6 ATOM 1601 CG ASN A215 29.256 14.419 38.2231.00 31.85 6 ATOM 1602 OD1ASN A215 29.976 14.179 37.2511.00 28.69 8 50ATOM 1603 ND2ASN A215 28.346 15.396 38.1161.00 31.00 7 ATOM 1604 N ASN A216 29.395 9.955 40.3641.00 15.65 7 ATOM 1605 CA ASN A216 29.543 9.014 41.4661.00 14.86 6 ATOM 1606 C ASN A216 28.733 9.469 42.6781.00 14.58 6 ATOM 1607 O ASN A216 29.202 9.375 43.8651.00 19.68 8 55ATOM 1608 CB ASN A216 31.049 8.93$ 41.8931.00 15.86 6 ATOM 1609 CG ASN A216 31.212 7.736 42.7991.00 19.49 6 ATOM 1610 OD1ASN A216 30.528 6.722 42.7241.00 15.44 B

ATOM 1611 ND2ASN A216 32.178 7.$41 43.7351.00 22.33 7 ATOM 1612 N GLY A217 27.452 9.740 42.4501.00 15.37 7 60ATOM 1613 CA GLY A217 26.528 10.146 43.4841.00 15.63 6 ATOM 1614 C GLY A217 25.397 9.108 43.6581.00 12.67 6 ATOM 1615 O GLY A217 25.274 8.223 42.8301.00 13.61 8 ATOM 1616 N TYR A218 24.763 9.167 44.8201.00 14.53 7 ATOM 1617 CA TYR A218 23.540 8.474 45.1471.00 13.47 6 65ATOM 1618 C TYR A218 22.573 9.467 45.7481.00 17.36 6 ATOM 1619 O TYR A218 23.042 10.313 46.5551.00 19.27 8 ATOM 1620 CB TXR A21$ 23.781 7.308 46.1611.00 11.47 6 ATOM 1621 CG TYR A218 24.809 6.356 45.5911.00 12.20 6 ATOM 1622 CD1 TYRA 218 26.208 6.488 45.773 1.0013.37 6 ATOM 1623 CD2 TYRA 218 24.426 5.277 44.820 1.0012.84 6 ATOM 1624 CE1 TYRA 218 27.084 5.638 45.230 1.0013.51 6 ATOM 1625 CE2 TYRA 218 25.300 4.451 44.233 1.0011.27 6 ATOM 1626 CZ TYRA 218 26.691 4.566 44.425 1.0011.33 6 ATOM 1627 OH TYRA 218 27.563 3.704 43.856 1.0014.19 8 ATOM 1628 N SERA 219 21.302 9.383 45.406 1.0013.70 7 ATOM 1629 CA SERA 219 20.345 10.29245.991 1.0012.43 6 ATOM 1630 C SERA 219 18.946 9.690 45.979 1.0012.57 6 10ATOM 1631 0 SERA 219 18.687 8.690 45.259 1.0011.67 8 ATOM 1632 CB SERA 219 20.380 11.57145.175 1.0016.94 6 ATOM 1633 OG SERA 219 19.543 11.40144.030 1.0027.04 8 ATOM 1634 N TYRA 220 18.080 10.23646.794 1.0011.86 7 ATOM 1635 CA TYRA 220 16.690 9.896 46.724 1.0011.51 6 15ATOM 1636 C TYRA 220 16.047 10.72245.635 1.0012.74 6 ATOM 1637 O TYRA 220 16.188 11.95345.508 1.0013.70 8 ATOM 1638 CB TYRA 220 16.005 10.33748.053 1.0011.07 6 ATOM 1639 CG TYRA 220 16.356 9.479 49.223 1.0015.23 6 ATOM 1640 CD1 TYRA 220 16.096 8.130 49.290 1.0012.19 6 20ATOM 1641 CD2 TYRA 220 16.970 10.06550.348 1.0020.94 6 ATOM 1642 CE1 TYRA 220 16.418 7.319 50.363 1.0017.15 6 ATOM 1643 CE2 TYRA 220 17.282 9.257 51.432 1.0020.63 6 ATOM 1644 CZ TYRA 220 17.013 7.927 51.455 1.0020.24 6 ATOM 1645 OH TYRA 220 17.330 7.134 52.548 1.0022.60 8 25ATOM 1646 N META 221 15.485 10.09844.923 1.0012.98 7 ATOM 1647 CA META 221 14.179 10.78643.985 1.0010.83 6 ATOM 1648 C META 221 12.794 10.19144.197 1.0010.00 6 ATOM 1649 O META 221 12.691 9.016 44.620 1.0011.94 8 ATOM 1650 CB META 221 14.581 10.67542.492 1.0012.31 6 30ATOM 1651 CG META 221 15.728 11.61142.190 1.0013.70 6 ATOM 1652 SD META 221 15.997 11.53040.390 1.0015.78 16 ATOM 1653 CE META 221 17.585 12.29240.256 1.0023.46 6 ATOM 1654 N SERA 222 11.723 10.90143.905 1.0010.92 7 ATOM 1655 CA SERA 222 10.357 10.42244.042 1.009.76 6 35ATOM 1656 C SERA 222 9.586 10.75142.758 1.0012.04 6 ATOM 1657 O SERA 222 9.755 11.82742.185 1.0015.35 B

ATOM 1658 CB SERA 222 9.609 11.10045.197 1.0014.86 6 ATOM 1659 OG SERA 222 10.216 10.86146.463 1.0013.78 B

ATOM 1660 N GLYA 223 8.779 9.812 42.394 1.0012.72 7 40ATOM 1661 CA GLYA 223 7.819 10.01441.264 1.0015.17 6 ATOM 1662 C GLYA 223 7.505 8.657 40.630 1.0010.31 6 ATOM 1663 0 GLYA 223 8.041 7.604 40.919 1.0011.61 8 ATOM 1664 N THRA 224 6.499 8.863 39.717 1.0010.35 7 ATOM 1665 CA THRA 224 6.175 7.685 38.909 1.008.89 6 45ATOM 1666 C THRA 224 7.383 7.335 38.027 1.009.98 6 ATOM 1667 O THRA 224 7.487 6.168 37.607 1.009.81 8 ATOM 1668 CB THRA 224 4.920 7.788 38.038 1.008.97 6 ATOM 1669 OG1 THRA 224 5.026 8.958 37.216 1.0011.03 8 ATOM 1670 CG2 THRA 224 3.671 7.909 38.950 1.009.27 6 50ATOM 1671 N SERA 225 8.317 8.243 37.735 1.0010.17 7 ATOM 1672 CA SERA 225 9.552 7.955 37.067 1.0011.35 6 ATOM 1673 C SERA 225 10.427 6.946 37.830 1.008.09 6 ATOM 1674 O SERA 225 11.258 6.285 37.163 1.008.80 8 ATOM 1675 CB SERA 225 10.482 9.186 36.881 1.0012.32 6 55ATOM 1676 OG SERA 225 9.832 10.04335.881 1.0013.46 8 ATOM 1677 N META 226 10.191 6.844 39.146 1.008.91 7 ATOM 1678 CA META 226 10.978 5.898 39.914 1.009.87 6 ATOM 1679 C META 226 10.277 4.558 40.071 1.008.01 6 ATOM 1680 O META 226 10.852 3.512 40.318 1.0010.42 B

60ATOM 1681 CB META 226 11.246 6.476 41.336 1.0010.18 6 ATOM 1682 CG META 226 12.310 7.569 41.381 1.009.98 6 ATOM 1683 SD META 226 11.911 9.112 40.574 1.0011.41 16 ATOM 1684 CE META 226 13.090 8.930 39.205 1.0013.09 6 ATOM 1685 N ALAA 227 8.939 4.581 39.923 1.009.49 7 65ATOM 1686 CA ALAA 227 8.136 3.349 40.019 1.009.59 6 ATOM 1687 C ALAA 227 8.327 2.524 38.724 1.008.92 6 ATOM 1688 O ALAA 227 8.449 1.279 38.759 1.009.60 8 ATOM 1689 CB ALAA 227 6.684 3.754 40.239 1.0012.00 6 ATOM 1690 N SER A 228 8.258 3.251 37.5981.00 8.46 7 ATOM 1691 CA SER A 228 8.366 2.564 36.2931.00 9.33 6 ATOM 1692 C SER A 228 9.597 1.685 36.1771.00 9.11 6 ATOM 1693 O SER A 228 9.393 0.525 35.7681.00 8.88 8 ATOM 1694 CB SER A 228 8.311 3.644 35.2221.00 8.72 6 ATOM 1695 OG SER A 228 8.326 3.035 33.8931.00 8.59 8 ATOM 1696 N PRO A 229 10.790 2.071 36.5691.00 9.43 7 ATOM 1697 CA PRO A 229 11.941 1.221 36.4201.00 9.18 6 ATOM 1698 C PRO A 229 11.901 -0.018 37.3121.00 9.20 6 10ATOM 1699 O PRO A 229 12.519 -1.041 37.0771.00 10.50 8 ATOM 1700 CB PRO A 229 13.198 2.065 36.7441.00 10.68 6 ATOM 1701 CG PRO A 229 12.614 3.303 37.4591.00 10.44 6 ATOM 1702 CD PRO A 229 11.196 3.448 36.8321.00 10.93 6 ATOM 1703 N HIS A 230 11.144 0.079 38.4591.00 8.14 7 15ATOM 1704 CA HIS A 230 10.984 -1.193 39.1941.00 8.04 6 ATOM 1705 C HIS A 230 10.199 -2.229 38.3981.00 8.61 6 ATOM 1706 0 HIS A 230 10.502 -3.422 38.3831.00 11.42 8 ATOM 1707 CB HIS A 230 10.245 -0.952 40.5671.00 8.82 6 ATOM 1708 CG HIS A 230 11.092 -0.269 41.6321.00 7.05 6 20ATOM 1709 ND1HIS A 230 11.161 1.109 41.7771.00 7.95 7 ATOM 1710 CD2HIS A 230 11.925 -0.847 42.5791.00 10.46 6 ATOM 1711 CE1HIS A 230 12.001 1.359 42.7911.00 11.65 6 ATOM 1712 NE2HIS A 230 12.464 0.188 43.2831.00 10.70 7 ATOM 1713 N VAL A 231 9.136 -1.757 37.7021.00 9.27 7 25ATOM 1714 CA VAL A 231 8.379 -2.623 36.8081.00 9.26 6 ATOM 1715 C VAL A 231 9.199 -3.052 35.6011.00 8.69 6 ATOM 1716 O VAL A 231 9.159 -4.237 35.2551.00 9.29 8 ATOM 1717 CB VAL A 231 7.078 -1.927 36.3891.00 8.06 6 ATOM 1718 CG1VAL A 231 6.310 -2.873 35.4441.00 12.86 6 30ATOM 1719 CG2VAL A 231 6.254 -1.603 37.6461.00 9.44 6 ATOM 1720 N ALA A 232 9.967 -2.118 35.0101.00 8.72 7 ATOM 1721 CA ALA A 232 10.810 -2.548 33.9001.00 9.54 6 ATOM 1722 C ALA A 232 11.885 -3.530 34.2911.00 9.30 6 ATOM 1723 0 ALA A 232 12.173 -4.505 33.5671.00 9.44 8 35ATOM 1724 CB ALA A 232 11.466 -1.262 33.2991.00 10.48 6 ATOM 1725 N GLY A 233 12.382 -3.408 35.5531.00 9.18 7 ATOM 1726 CA GLY A 233 13.367 -4.396 36.0191.00 9.95 6 ATOM 1727 C GLY A 233 12.770 -5.794 36.2141.00 8.05 6 ATOM 1728 O GLY A 233 13.315 -6.802 35.8701.00 9.80 8 40ATOM 1729 N LEU A 234 11.543 -5.846 36.7991.00 8.56 7 ATOM 1?30 CA LEU A 234 10.797 -7.116 36.8611.00 9.22 6 ATOM 1731 C LEU A 234 10.564 -7.685 35.4521.00 8.93 6 ATOM 1732 O LEU A 234 10.703 -8.869 35.2571.00 11.08 8 ATOM 1733 CB LEU A 234 9.502 -6.924 37.6731.00 10.61 6 45ATOM 1734 CG LEU A 234 8.527 -8.118 37.6751.00 10.54 6 ATOM 1735 CD1LEU A 234 9.338 -9.334 38.1721.00 10.76 6 ATOM 1736 CD2LEU A 234 7.332 -7.778 38.5161.00 11.53 6 ATOM 1737 N ALA A 235 10.192 -6.794 34.5141.00 8.31 7 ATOM 1738 CA ALA A 235 9.995 -7.321 33.1591.00 9.02 6 50ATOM 1739 C ALA A 235 11.282 -7.907 32.6381.00 9.50 6 ATOM 1740 O ALA A 235 11.236 -8.925 31.9051.00 9.26 8 ATOM 1741 CB ALA A 235 9.452 -6.193 32.2831.00 8.43 6 ATOM 1742 N ALA A 236 12.423 -7.302 32.9831.00 9.58 7 ATOM 1743 CA ALA A 236 13.686 -7.893 32.4811.00 10.25 6 55ATOM 1744 C ALA A 236 13.951 -9.229 33.2001.00 9.83 6 ATOM 1745 0 ALA A 236 14.536 -10.13432.5611.00 11.26 8 ATOM 1746 CB ALA A 236 14.866 -6.960 32.7771.00 11.37 6 ATOM 1747 N LEU A 237 13.560 -9.388 34.4551.00 10.12 7 ATOM 1748 CA LEU A 237 13.759 -10.69135.1021.00 11.28 6 60ATOM 1749 C LEU A 23'7 12.950 -11.74934.3871.00 11.62 6 ATOM 1750 O LEU A 237 13.438 -12.83834.0621.00 11.77 8 ATOM 1751 CB LEU A 237 13.237 -10.61536.5771.00 9.89 6 ATOM 1752 CG LEU A 237 14.114 -9.812 37.5281.00 10.68 6 ATOM 1753 CD1LEU A 237 13.489 -9.905 38.9361.00 10.83 6 65ATOM 1754 CD2LEU A 237 15.587 -10.20637.5431.00 12.79 6 ATOM 1755 N LEU A 238 11.724 -11.38634.0071.00 10.46 7 ATOM 1756 CA LEU A 238 10.866 -12.31333.2581.00 10.34 6 ATOM 1757 C LEU A 238 11.442 -12.56131.8561.00 11.39 6 ATOM 1758 O LEUA 238 11.368 -13.72731.401 1.0011.82 B

ATOM 1759 CB LEUA 238 9.446 -11.74733.253 1.0011.04 ATOM 1760 CG LEUA 238 8.748 -11.65234.596 1.009.18 ATOM 1761 CD1 LEUA 238 7.624 -10.61034.653 1.0013.12 ATOM 1762 CD2 LEUA 238 8.198 -13.03734.992 1.0014.13 ATOM 1763 N ALAA 239 11.985 -11.57031.187 1.0011.41 ATOM 1764 CA ALAA 239 12.536 -11.83429.836 1.0011.12 ATOM 1765 C ALAA 239 13.707 -12.81029.917 1.0013.22 ATOM 1766 0 ALAA 239 13.904 -13.58428.964 1.0014.69 l0 ATOM 1767 CB ALAA 239 13.017 -10.50729.271 1.0011.43 ATOM 1768 N SERA 240 14.399 -12.83331.034 1.0010.04 ATOM 1769 CA SERA 240 15.560 -13.70631.190 1.0011.57 ATOM 1770 C SERA 240 15.069 -15.15831.346 1.0011.74 ATOM 1771 O SERA 240 15.978 -16.00731.212 1.0017.10 B

ATOM 1772 CB SERA 240 16.467 -13.25332.339 1.0015.20 ATOM 1773 OG SERA 240 15.907 -13.48733.636 1.0015.38 B

ATOM 1774 N GLNA 241 13.796 -15.39031.539 1.0012.24 ATOM 1775 CA GLNA 241 13.311 -16.76531.686 1.0013.26 ATOM 1776 C GLNA 241 12.864 -17.29830.311 1.0016.69 ATOM 1777 O GLNA 241 12.259 -18.38130.257 1.0017.39 ATOM 1778 CB GLNA 241 12.171 -16.79432.671 1.0012.49 ATOM 1779 CG GLNA 241 12..632 -16.40834.116 1.0014.22 ATOM 1780 CD GLNA 241 11.364 -16.35134.927 1.0014.64 ATOM 1781 OE1 GLNA 241 10.413 -15.56934.925 1.0015.36 ATOM 1782 NE2 GLNA 241 11.248 -17.40935.756 1.0014.32 ATOM 1783 N GLYA 242 13.086 -16.40429.347 1.0016.05 ATOM 1784 CA GLYA 242 12.689 -16.81227.982 1.0017.81 ATOM 1785 C GLYA 242 11.249 -16.54927.658 1.0018.23 ATOM 1786 O GLYA 242 10.619 -17.05426.710 1.0019.39 ATOM 1787 N LYSA 243 10.538 -15.77828.476 1.0015.28 ATOM 1788 CA LYSA 243 9.137 -15.45828.344 1.0012.91 ATOM 1789 C LYSA 243 9.010 -14.33627.290 1.0013.52 ATOM 1790 O LYSA 243 9.837 -13.40827.296 1.0016.08 ATOM 1791 CB LYSA 243 8.541 -14.99529.664 1.0015.99 ATOM 1792 CG LYSA 243 8.278 -16.18530.633 1.0016.79 ATOM 1793 CD LYSA 243 8.070 -15.68732.070 1.0021.08 ATOM 2794 CE LYSA 243 8.016 -17.01632.871 1.0025.90 ATOM 1795 NZ LYSA 243 8.276 -16.82934.309 1.0025.09 ATOM 1796 N ASNA 244 7.971 -14.39626.458 1.0012.59 ATOM 1797 CA ASNA 244 7.851 -13.36025.428 1.0012.14 ATOM 1798 C ASNA 244 7.060 -12.17226.019 1.0012.86 ATOM 1799 O ASNA 244 6.643 -12.19527.204 1.0012.30 ATOM 1800 CB ASNA 244 7.141 -13.96924.225 1.0011.92 ATOM 1801 CG ASNA 244 5.733 -14.36324.487 1.0014.11 ATOM 1802 OD1 ASNA 244 4.907 -13.89125.246 1.0014.17 ATOM 1803 ND2 ASNA 244 5.380 -15.45023.740 1.0025.93 ATOM 1804 N ASNA 245 6.779 -11.15825.223 1.0011.75 ATOM 1805 CA ASNA 245 6.231 -9.925 25.803 1.0012.32 ATOM 1806 C ASNA 245 4.838 -10.18526.373 1.0011.73 ATOM 1807 O ASNA 245 4.488 -9.607 27.389 1.0011.85 B

ATOM 1808 CB ASNA 245 6.266 -8.706 24.917 1.009.82 ATOM 1809 CG ASNA 245 5.639 -8.895 23.503 1.0011.05 ATOM 1810 OD1 ASNA 245 5.182 -9.975 23.141 1.0012.49 ATOM 1811 ND2 ASNA 245 5.668 -7.759 22.832 1.0011.22 ATOM 1812 N VALA 246 4.041 -10.93825.608 1.0012.28 ATOM 1813 CA VALA 246 2.701 -11.23126.102 1.0011.50 ATOM 1814 C VALA 246 2.798 -11.93227.464 1.0012.02 ATOM 1815 0 VALA 246 2.046 -11.54128.399 1.0013.42 ATOM 1816 CB VALA 246 1.936 -12.08825.084 1.0013.60 ATOM 1817 CG1 VALA 246 0.562 -12.52725.579 1.0017.63 ATOM 1818 CG2 VALA 246 1.693 -11.26623.802 1.0015.00 ATOM 1819 N GLNA 247 3.644 -12.91327.577 1.0013.17 ATOM 1820 CA GLNA 247 3.880 -13.68228.797 1.0012.37 ATOM 1821 C GLNA 247 4.332 -12.76729.958 1.0013.38 ATOM 1822 O GLNA 247 3.853 -12.92231.081 1.0013.69 ATOM 1823 CB GLNA 247 4.936 -14.77828.529 1.0014.20 ATOM 1824 CG GLNA 24? 4.208 -15.91127.719 1.0016.06 ATOM 1825 CD GLNA 247 5.269 -16.94127.390 1.0019.34 ATOM 1826 OE1 GLNA 247 6.402 -16.70627.0421.00 16.30 B

ATOM 1827 NE2 GLNA 247 4.878 -18.22427.5351.00 34.55 7 ATOM 1828 N ILEA 248 5.310 -11.93729.6161.00 11.49 7 ATOM 1829 CA ILEA 248 5.861 -10.99730.6461.00 9.94 6 5 ATOM 1830 C ILEA 248 4.771 -10.09931.2091.00 13.72 6 ATOM 1831 O ILEA 248 4.620 -9.931 32.4361.00 10.62 8 ATOM 1832 CB ILEA 248 6.992 -10.16930.0021.00 9.93 6 ATOM 1833 CG1 ILEA 248 8.162 -11.06729.6871.00 12.15 6 ATOM 1834 CG2 ILEA 248 7.433 -9.060 30.9901.00 12.79 6 10ATOM 1835 CDl ILEA 248 9.257 -10.33628.8941.00 12.72 6 ATOM 1836 N ARGA 249 4.049 -9.408 30.3101.00 11.43 7 ATOM 1837 CA ARGA 249 3.025 -8.484 30.7671.00 10.98 6 ATOM 1838 C ARGA 249 1.907 -9.207 31.4991.00 10.67 6 ATOM 1839 O ARGA 249 1.515 -8.692 32.5851.00 12.16 8 15ATOM 1840 CB ARGA 249 2.523 -7.745 29.4861.00 12.64 6 ATOM 1841 CG ARGA 249 1.422 -6.790 30.0091.00 14.83 6 ATOM 1842 CD ARGA 249 0.941 -5.857 28.8931.00 11.70 6 ATOM 1843 NE ARGA 249 0.026 -4.852 29.4851.00 12.16 7 ATOM 1844 CZ ARGA 249 -0.233 -3.681 28.9091.00 17.32 6 20ATOM 1845 NH1 ARGA 249 0.242 -3.351 27.6971.00 12.81 7 ATOM 1846 NH2 ARGA 249 -1.008 -2.825 29.5821.00 17.64 7 ATOM 1847 N GLNA 250 1.444 -10.38831.0871.00 12.08 7 ATOM 1848 CA GLNA 250 0.444 -11.14631.8011.00 12.15 6 ATOM 1849 C GLNA 250 0.953 -11.52733.1831.00 12.65 6 25ATOM 1850 O GLNA 250 0.181 -11.41034.1541.00 13.71 8 ATOM 1851 CB GLNA 250 0.049 -12.40130.9821.00 13.50 6 ATOM 1852 CG GLNA 250 -1.139 -13.17731.5201.00 25.34 6 ATOM 1853 CD GLNA 250 -1.654 -14.04230.3511.00 34.38 6 ATOM 1854 OE1 GLNA 250 -0.926 -14.23129.3791.00 40.16 8 30ATOM 1855 NE2 GLNA 250 -2.883 -14.52530.4531.00 47.06 7 ATOM 1856 N ALAA 251 2.236 -11.90633.2781.00 10.23 7 ATOM 1857 CA ALAA 251 2.706 -12.32334.6201.00 12.97 6 ATOM 1858 C ALAA 251 2.739 -11.13535.5351.00 12.84 6 ATOM 1859 O ALAA 251 2.312 -11.21236.7171.00 12.96 8 35ATOM 1860 CB ALAA 251 4.077 -12.95734.4701.00 14.38 6 ATOM 1861 N ILEA 252 3.171 -9.986 35.0451.00 11.02 7 ATOM 1862 CA ILEA 252 3.233 -8.795 35.9471.00 10.23 6 ATOM 1863 C ILEA 252 1.845 -8.365 36.4061.00 10.23 6 ATOM 1864 O ILEA 252 1.633 -8.028 37.6161.00 13.67 8 40ATOM 1865 CB ILEA 252 3.878 -7.669 35.1121.00 12.15 6 ATOM 1866 CG1 ILEA 252 5.396 -7.885 34.9811.00 11.82 6 ATOM 1867 CG2 ILEA 252 3.648 -6.337 35.8431.00 13.44 6 ATOM 1868 CD1 ILEA 252 6.044 -6.969 33.9491.00 13.14 6 ATOM 1869 N GLUA 253 0.879 -8.376 35.4921.00 12.32 7 45ATOM 1870 CA GLUA 253 -0.446 -7.831 35.8331.00 13.24 6 ATOM 1871 C GLUA 253 -1.237 -8.841 36.6401.00 12.78 6 ATOM 1872 0 GLUA 253 -1.895 -8.507 37.6211.00 15.59 8 ATOM 1873 CB GLUA 253 -1.189 -7.514 34.5071.00 11.47 6 ATOM 1874 CG GLUA 253 -0.638 -6.220 33.9081.00 11.43 6 50ATOM 1875 CD GLUA 253 -1.256 -5.936 32.5271.00 15.21 6 ATOM 1876 OE1 GLUA 253 -1.899 -6.803 31.9261.00 18.21 8 ATOM 1877 OE2 GLUA 253 -1.005 -4.753 32.2061.00 17.23 8 ATOM 1878 N GLNA 254 -1.220 -10.13436.2121.00 12.34 7 ATOM 1879 CA GLNA 254 -2.095 -11.12236.8421.00 13.43 6 55ATOM 1880 C GLNA 254 -1.626 -11.50138.2221.00 13.73 6 ATOM 1881 0 GLNA 254 -2.499 -11.96639.0241.00 19.17 8 ATOM 1882 CB GLNA 254 -2.151 -12.36435.9661.00 14.03 6 ATOM 1883 CG GLNA 254 -2.890 -12.15934.6511.00 15.03 6 ATOM 1884 CD GLNA 254 -4.361 -11.90634.8851.00 16.67 6 60ATOM 1885 OE1 GLNA 254 -5.113 -12.66535.5201.00 18.71 8 ATOM 1886 NE2 GLNA 254 -4.734 -10.73834.3461.00 19.67 7 ATOM 1887 N THRA 255 -0.340 -11.32338.5701.00 12.59 7 ATOM 1888 CA THRA 255 0.078 -11.66639.9381.00 12.50 6 ATOM 1889 C THRA 255 0.218 -10.48040.8761.00 11.92 6 65ATOM 1890 0 THRA 255 0.652 -10.69841.9991.00 13.55 8 ATOM 1891 CB THRA 255 1.449 -12.36739.9191.00 11.55 6 ATOM 1892 OG1 THRA 255 2.490 -11.50739.3571.00 12.19 8 ATOM 1893 CG2 THRA 255 1.423 -13.63539.0541.00 12.96 6 WO 00/22103 PC'T/DK99/00542 ATOM 1894 N ALA A 256 -0.211 -9.309 40.4171.00 12.42 7 ATOM 1895 CA ALA A 256 0.004 -8.131 41.3071.00 11.98 6 ATOM 1896 C ALA A 256 -0.832 -8.313 42.5631.00 13.19 6 ATOM 1897 O ALA A 256 -1.957 -8.835 42.5051.00 15.40 8 ATOM 1898 CB ALA A 256 -0.438 -6.931 40.5021.00 13.07 6 ATOM 1899 N ASP A 257 -0.266 -7.824 43.6831.00 10.82 7 ATOM 1900 CA ASP A 257 -1.023 -7.893 44.9541.00 11.39 6 ATOM 1901 C ASP A 257 -2.151 -6.879 44.9211.00 12.69 6 ATOM 1902 O ASP A 257 -1.949 -5.658 44.6721.00 12.22 8 10ATOM 1903 CB ASP A 257 -0.048 -7.519 46.0641.00 12.06 6 ATOM 1904 CG ASP A 257 0.966 -8.615 46.3391.00 14.17 6 ATOM 1905 ODlASP A 257 0.623 -9.774 46.0831.00 17.70 8 ATOM 1906 OD2ASP A 257 2.038 -8.290 46.8931.00 18.13 8 ATOM 1907 N LYS A 258 -3.323 -7.323 45.3411.00 12.79 7 15ATOM 1908 CA LYS A 258 -4.489 -6.445 45.3581.00 13.57 6 ATOM 1909 C LYS A 258 -4.583 -5.594 46.5921.00 14.49 6 ATOM 1910 0 LYS A 258 -5.543 -5.711 47.3891.00 16.26 8 ATOM 1911 CB LYS A 258 -5.790 -7.213 45.0521.00 17.86 6 ATOM 1912 CG LYS A 258 -5.563 -7.937 43.7061.00 22.99 6 20ATOM 1913 CD LYS A 258 -6.836 -8.271 42.9541.00 27.90 6 ATOM 1914 CE LYS A 258 -6.527 -9.082 41.7071.00 24.57 6 ATOM 1915 NZ LYS A 258 -5.879 -8.283 40.6051.00 25.24 7 ATOM 1916 N ILE A 259 -3.678 -4.648 46.7561.00 15.10 7 ATOM 1917 CA ILE A 259 -3.576 -3.775 47.8981.00 13.74 6 25ATOM 1918 C ILE A 259 -4.677 -2.717 47.8671.00 13.04 6 ATOM 1919 O ILE A 259 -5.360 -2.579 46.8451.00 12.63 8 ATOM 1920 CB ILE A 259 -2.175 -3.096 47.9811.00 14.87 6 ATOM 1921 CG1ILE A 259 -1.974 -2.187 46.7641.00 14.73 6 ATOM 1922 CG2ILE A 259 -1.086 -4.132 48.1531.00 13.52 6 30ATOM 1923 CD1ILE A 259 -0.796 -1.246 46.9011.00 14.29 6 ATOM 1924 N SER A 260 -4.840 -1.987 48.9851.00 13.66 7 ATOM 1925 CA SER A 260 -5.820 -0.905 48.9281.00 13.91 6 ATOM 1926 C SER A 260 -5.545 0.022 47.7601.00 14.86 6 ATOM 1927 O SER A 260 -4.392 0.338 47.4151.00 15.35 8 35ATOM 1928 CB SER A 260 -5.652 -0.158 50.2711.00 23.45 6 ATOM 1929 OG SER A 260 -6.523 0.961 50.2641.00 32.81 8 ATOM 1930 N GLY A 261 -6.615 0.415 47.0651.00 14.79 7 ATOM 1931 CA GLY A 261 -6.451 1.204 45.8531.00 15.91 6 ATOM 1932 C GLY A 261 -6.756 0.360 44.6171.00 12.68 6 40ATOM 1933 0 GLY A 26:L-6.863 0.965 43.5341.00 12.20 8 ATOM 1934 N THR A 262 -6.655 -0.924 44.7051.00 12.44 7 ATOM 1935 CA THR A 262 -6.915 -1.796 43.5371.00 10.57 6 ATOM 1936 C THR A 262 -8.331 -1.542 43.0301.00 13.90 6 ATOM 1937 O THR A 262 -9.301 -1.632 43.8191.00 15.95 8 45ATOM 1938 CB THR A 262 -6.699 -3.286 43.8401.00 13.48 6 ATOM 1939 OG1THR A 262 -5.331 -3.420 44.2291.00 14.15 8 ATOM 1940 CG2THR A 262 -6.959 -4.137 42.6171.00 16.51 6 ATOM 1941 N GLY A 263 -8.396 -1.179 41.7471.00 11.09 7 ATOM 1942 CA GLY A 263 -9.735 -0.902 41.1521.00 12.45 6 50ATOM 1943 C GLY A 263 -10.071 0.568 41.1581.00 11.46 6 ATOM 1944 O GLY A 263 -10.990 1.093 40.4641.00 14.08 8 ATOM 1945 N THR A 264 -9.309 1.422 41.8721.00 10.76 7 ATOM 1946 CA THR A 264 -9.488 2.859 41.9331.00 10.53 6 ATOM 1947 C THR A 264 -8.266 3.602 41.4011.00 11.33 6 55ATOM 1948 O THR A 264 -8.356 4.471 40.5181.00 13.25 8 ATOM 1949 CB ATHRA 264 -9.810 3.214 43.4000.50 13.28 6 ATOM 1950 OG1ATHR A 264 -10.941 2.511 43.8970.50 13.13 8 ATOM 1951 CG2ATHRA 264 -9.919 4.711 43.4360.50 8.33 6 ATOM 1952 CB BTHRA 264 -9.844 3.467 43.3080.50 11.69 6 60ATOM 1953 OG1BTHRA 264 -8.956 2.998 44.3440.50 11.80 8 ATOM 1954 CG2BTHRA 264 -11.253 3.162 43.7240.50 10.13 6 ATOM 1955 N ASN A 265 -7.080 3.363 42.0001.00 10.66 7 ATOM 1956 CA ASN A 265 -5.841 4.057 41.6121.00 10.59 6 ATOM 1957 C ASN A 265 -5.059 3.298 40.5731.00 11.52 6 65ATOM 1958 O ASN A 265 -4.186 3.892 39.9061.00 11.50 8 ATOM 1959 CB ASN A 265 -4.983 4.241 42.8591.00 11.37 6 ATOM 1960 CG ASN A 265 -5.590 5.258 43.8261.00 12.28 6 ATOM 1961 OD1ASN A 265 -6.418 6.059 43.4161.00 15.68 8 WO 00/22103 PCTlDK99/00542 ATOM 1962 ND2 ASNA 265 -5.153 5.175 45.083 1.00 16.82 7 ATOM 1963 N PHEA 266 -5.368 2.029 40.370 1.00 12.07 7 ATOM 1964 CA PHEA 266 -4.728 1.230 39.337 1.00 13.47 6 ATOM 1965 C PHEA 266 -5.576 -0.003 39.144 1.00 11.98 6 ATOM 1966 0 PHEA 266 -6.414 -0.342 40.025 1.00 13.68 8 ATOM 1967 CB PHEA 266 -3.273 0.833 39.743 1.00 11.51 6 ATOM 1968 CG PHEA 266 -3.191 0.624 41.228 1.00 10.99 6 ATOM 1969 CD1 PHEA 266 -2.709 1.603 42.034 1.00 12.36 6 ATOM 1970 CD2 PHEA 266 -3.617 -0.589 41.768 1.00 12.87 6 10ATOM 1971 CE1 PHEA 266' -2.646 1.489 43.437 1.00 15.18 6 ATOM 1972 CE2 PHEA 266 -3.548 -0.720 43.157 1.00 13.50 6 ATOM 1973 CZ PHEA 266 -3.086 0.299 43.954 1.00 13.91 6 ATOM 1974 N LYSA 267 -5.481 -0.713 38.018 1.00 10.86 7 ATOM 1975 CA LYSA 267 -6.372 -1.827 37.757 1.00 12.35 6 15ATOM 1976 C LYSA 267 -5.996 -3.143 38.415 1.00 11.29 6 ATOM 1977 O LYSA 267 -6.827 -3.835 39.018 1.00 14.13 8 ATOM 1978 CB LYSA 267 -6.427 -2.056 36.234 1.00 10.21 6 ATOM 1979 CG LYSA 267 -7.269 -3.230 35.800 1.00 10.69 6 ATOM 1980 CD LYSA 267 -7.434 -3.314 34.277 1.00 17.41 6 20ATOM 1981 CE LYSA 267 -8.125 -4.645 33.961 1.00 20.47 6 ATOM 1982 NZ LYSA 267 -9.590 -4.515 34.312 1.00 28.85 7 ATOM 1983 N TYRA 268 -4.703 -3.507 38.377 1.00 10.94 7 ATOM 1984 CA TYRA 268 -4.306 -4.845'38.774 1.00 10.54 6 ATOM 1985 C TYRA 268 -3.780 -4.934 40.185 1.00 12.48 6 25ATOM 1986 0 TYRA 268 -4.004 -5.966 40.828 1.00 14.47 8 ATOM 1987 CB TYRA 268 -3.247 -5.379 37.737 1.00 11.99 6 ATOM 1988 CG TYRA 268 -3.869 -5.582 36.354 1.00 12.59 6 ATOM 1989 CD1 TYRA 268 -4.729 -6.636 36.098 1.00 17.30 6 ATOM 1990 CD2 TYRA 268 -3.567 -4.713 35.315 1.00 11.77 6 30ATOM 1991 CE1 TYRA 268 -5.286 -6.819 34.836 1.00 16.83 6 ATOM 1992 CE2 TYRA 268 -4.127 -4.878 34.044 1.00 14.50 6 ATOM 1993 CZ TYRA 268 -4.975 -5.940 33.842 1.00 16.27 6 ATOM 1994 OH TYRA 268 -5.578 -6.179 32.594 1.00 16.50 8 ATOM 1995 N GLYA 269 -2.951 -3.959 40.589 1.00 11.21 7 35ATOM 1996 CA GLYA 269 -2.363 -4.068 41.950 1.00 11.42 6 ATOM 1997 C GLYA 269 -0.884 -3.640 41.935 1.00 12.63 6 ATOM 1998 0 GLYA 269 -0.410 -2.979 40.981 1.00 12.33 8 ATOM 1999 N LYSA 270 -0.250 -3.975 43.039 1.00 10.84 7 ATOM 2000 CA LYSA 270 1.158 -3.689 43.298 1.00 10.02 6 40ATOM 2001 C LYSA 270 1.989 -4.849 42.786 1.00 10.57 6 ATOM 2002 O LYSA 270 1.759 -6.030 43.116 1.00 10.24 8 ATOM 2003 CB LYSA 270 1.355 -3.550 44.835 1.00 9.80 6 ATOM 2004 CG LYSA 270 2.838 -3.440 45.234 1.00 10.47 6 ATOM 2005 CD LYSA 270 2.766 -3.574 46.792 1.00 13.93 6 45ATOM 2006 CE LYSA 270 4.141 -3.882 47.323 1.00 13.15 6 ATOM 2007 NZ LYSA 270 4.141 -4.146 48.814 1.00 12.38 7 ATOM 2008 N ILEA 271 3.068 -4.553 41.981 1.00 9.21 7 ATOM 2009 CA ILEA 271 3.827 -5.684 41.486 1.00 10.54 6 ATOM 2010 C ILEA 271 4.345 -6.567 42.615 1.00 10.27 6 50ATOM 2011 O ILEA 271 4.722 -6.097 43.676 1.00 10.76 8 ATOM 2012 CB ILEA 271 5.015 -5.309 40.579 1.00 9.40 6 ATOM 2013 CGl ILEA 271 5.942 -4.342 41.302 1.00 10.70 6 ATOM 2014 CG2 ILEA 271 4.462 -4.651 39.299 1.00 12.37 6 ATOM 2015 CD1 ILEA 271 7.353 -4.230 40.661 1.00 11.12 6 55ATOM 2016 N ASNA 272 4.358 -7.864 42.304 1.00 11.32 7 ATOM 2017 CA ASNA 272 4.927 -8.895 43.187 1.00 11.90 6 ATOM 2018 C ASNA 272 5.887 -9.781 42.379 1.00 10.24 6 ATOM 2019 O ASNA 272 5.512 -10.58241.534 1.00 11.33 8 ATOM 2020 CB ASNA 272 3.791 -9.719 43.774 1.00 10.81 6 60ATOM 2021 CG ASNA 272 4.423 -10.70344.785 1.00 13.73 6 ATOM 2022 OD1 ASNA 272 5.449 -11.32444.563 1.00 11.13 8 ATOM 2023 ND2 ASNA 272 3.754 -10.77245.915 1.00 15.82 7 ATOM 2024 N SERA 273 7.192 -9.406 42.462 1.00 11.18 7 ATOM 2025 CA SERA 273 8.176 -10.05441.573 1.00 9.11 6 65ATOM 2026 C SERA 273 8.243 -11.57541.796 1.00 10.89 6 ATOM 2027 O SERA 273 8.463 -12.25640.830 1.00 11.77 8 ATOM 2028 CB SERA 273 9.575 -9.501 41.899 1.00 12.03 6 ATOM 2029 OG SERA 273 9.574 -8.068 41.685 1.00 11.29 8 ATOM 2030 N ASNA 274 8.098 -11.96843.092 1.0011.68 ATOM 2031 CA ASNA 274 8.221 -13.41743.328 1.0013.05 ATOM 2032 C ASNA 274 7.075 -14.19542.716 1.0011.54 ATOM 2033 O ASNA 274 7.388 -15.22842.085 1.0014.09 ATOM 2034 CB ASNA 274 8.376 -13.67844.846 1.0013.98 ATOM 2035 CG ASNA 274 8.826 -15.14945.061 I.0013.57 ATOM 2036 OD1 ASNA 274 9.757 -15.64044.474 1.0016.37 ATOM 2037 ND2 ASNA 274 8.018 -15.69045.988 1.0019.49 ATOM 2038 N LYSA 275 5.882 -13.69942.841 1.0012.00 10ATOM 2039 CA LYSA 275 4.726 -14.36642.195 1.0011.46 ATOM 2040 C LYSA 275 4.874 -14.29940.667 1.0012.56 ATOM 2041 0 LYSA 275 4.574 -15.28840.002 1.0012.54 ATOM 2042 CB LYSA 275 3.383 -13.89642.682 1.0014.49 ATOM 2043 CG LYSA 275 3.025 -13.90244.182 1.0017.77 15ATOM 2044 CD LYSA 275 1.573 -13.45144.400 1.0021.72 ATOM 2045 CE LYSA 275 1.152 -12.01544.481 1.0028.84 ATOM 2046 NZ LYSA 275 -0.267 -11.49644.363 1.0026.58 ATOM 2047 N ALAA 276 5.310 -13.11840.185 1.0012.51 ATOM 2048 CA ALAA 276 5.385 -13.05938.706 1.0010.12 20ATOM 2049 C ALAA 276 6.375 -14.01438.107 1.0010.25 ATOM 2050 0 ALAA 276 6.048 -14.63537.070 1.0010.48 ATOM 2057.CB ALAA 276 5.743 -11.61738.304 1.0011.31 ATOM 2052 N VALA 277 7.553 -14.22638.736 1.0011.74 ATOM 2053 CA VALA 277 8.541 -15.04138.026 1.0010.78 25ATOM 2054 C VALA 277 8.167 -16.51838.194 1.0014.27 ATOM 2055 O VALA 277 8.780 -17.29837.504 1.0013.97 ATOM 2056 CB VALA 277 9.970 -14.84238.526 1.0012.43 ATOM 2057 CG1 VALA 277 10.448 -13.42538.246 1.0012.15 ATOM 2058 CG2 VALA 277 10.184 -15.15140.012 1.0016.20 30ATOM 2059 N ARGA 278 7.190 -16.80939.071 1.0012.39 ATOM 2060 CA ARGA 278 6.784 -18.22339.175 1.0015.11 ATOM 2061 C ARGA 278 5.497 -18.50438.446 1.0016.59 ATOM 2062 0 ARGA 278 5.089 -19.68938.350 1.0020.78 ATOM 2063 CB ARGA 278 6.542 -18.58740.655 1.0015.94 35ATOM 2064 CG ARGA 278 7.824 -18.42241.383 1.0017.42 ATOM 2065 CD ARGA 278 7.595 -18.57242.905 1.0015.96 ATOM 2066 NE ARGA 278 8.833 -18.23043.559 1.0019.66 ATOM 2067 CZ ARGA 278 9.900 -18.90143.897 1.0024.54 ATOM 2068 NH1 ARGA 278 9.966 -20.22343.611 1.0023.93 40ATOM 2069 NH2 ARGA 278 10.918 -18.30544.529 1.0026.20 ATOM 2070 N TYRA 279 4.913 -17.49037.831 1.0012.78 ATOM 2071 CA TYRA 279 3.644 -17.62637.129 1.0014.37 ATOM 2072 C TYRA 279 3.850 -18.37835.829 1.0024.50 ATOM 2073 0 TYRA 279 4.901 -18.16535.189 1.0024.68 45ATOM 2074 CB TYRA 279 3.005 -16.27736.932 1.0013.65 ATOM 2075 CG TYRA 279 1.693 -16.10536.231 1.0013.86 ATOM 2076 CD1 TYRA 279 0.452 -16.18936.858 1.0014.68 ATOM 2077 CD2 TYRA 279 1.736 -15.87234.847 1.0015.48 ATOM 2078 CE1 TYRA 279 -0.683 -16.03736.103 1.0015.73 50ATOM 2079 CE2 TYRA 279 0.575 -15.70234.121 1.0014.88 ATOM 2080 CZ TYRA 279 -0.652 -15.77034.740 1.0015.75 ATOM 2081 OH TYRA 279 -1.834 -15.61234.089 1.0017.03 B

ATOM 2082 OT TYRA 279 3.000 -19.25835.525 1.0024.50 ATOM 2083 C1 GLLA 296 -3.949 0.135 29.717 1.0017.61 55ATOM 2084 C2 GLLA 296 -4.024 1.580 29.221 1.0016.18 ATOM 2085 C3 GLLA 296 -5.461 2.100 29.213 1.0018.89 ATOM 2086 O1 GLLA 296 -2.578 -0.308 29.529 1.0015.42 ATOM 2087 02 GLLA 296 -3.163 2.400 30.001 1.0015.61 ATOM 2088 03 GLLA 296 -5.525 3.473 28.806 1.0023.45 60ATOM 2089 CA WATA 1 25.973 -1.842 43.443 1.0011.05 ATOM 2090 CA WATA 2 25.647 13.399 23.751 1.0017.24 ATOM 2091 NA WATA 3 -1.258 1.535 28.929 1.0012.14 ATOM 2092 OWO WATW 4 16.838 2.112 43.195 1.009.07 ATOM 2093 OWO WATW 5 13.085 6.361 31.187 1.0010.24 65ATOM 2094 OWO WATW 6 18.887 0.537 42.447 1.0010.28 ATOM 2095 OWO WATW 7 14.445 5.591 39.775 1.0010.40 ATOM 2096 OWO WATW 8 14.210 1.611 47.107 1.0010.59 ATOM 2097 OWO WATW 9 14.918 3.839 48.721 1.0010.61 ATOM 2098 OWO WATW 10 10.698 -1.779 53.649 1.0010.77 8 ATOM 2099 OWO WATW 11 -1.751 5.117 40.655 1.0010.86 8 ATOM 2100 OWO WATW 1.2 14.945 0.687 44.697 1.0010.99 8 ATOM 2101 OWO WATW 13 -0.327 4.307 42.978 1.0011.25 8 ATOM 2102 OWO WATW 14 4.023 -3.345 27.168 1.0011.76 8 ATOM 2103 OWO WATW 15 3.256 -8.810 39.822 1.0011.88 8 ATOM 2104 OWO WATW 16 18.664 7.646 28.905 1.0012.48 8 ATOM 2105 OWO WATW 17 29.275 -1.039 37.125 1.0012.57 8 ATOM 2106 OWO WATW 18 20.255 15.290 29.790 1.0012.74 8 10ATOM 2107 OWO WATW 19 10.140 -8.862 25.520 1.0012.92 8 ATOM 2108 OWO WATW 20 -1.402 0.794 22.084 1.0013.07 8 ATOM 2109 OWO WATW 21 5.723 -0.410 49.244 1.0013.11 8 ATOM 2110 OWO WATW 22 11.383 -11.54549.015 1.0013.37 8 ATOM 2111 OWO WATW 23 32.574 -0.530 38.835 1.0013.78 8 15ATOM 2112 OWO WATW 24 6.840 6.391 14.824 1.0013.94 8 ATOM 2113 OWO WATW 25 4.270 -7.073 46.186 1.0014.15 8 ATOM 2114 OWO WATW 26 16.914 -1.439 53.452 1.0014.17 8 ATOM 2115 OWO WATW 27 25.377 -6.748 38.454 1.0014.66 8 ATOM 2116 OWO WATW 28 16.613 -9.407 30.953 1.0014.93 8 20ATOM 2117 OWO WATW 29 3.879 7.957 30.204 1.0014.99 8 ATOM 2118 OWO WATW 30 11.580 -4.612 12.085 1.0015.06 8 ATOM 2119 OWO WATW 31 8.559 -11.11022.717 1.0015.31 8 ATOM 2120 OWO WATW 32 31.655 -2.825 39.954 1.0015.39 8 ATOM 2121 OWO WATW 33 5.355 5.671 12.382 1.0015.85 8 25ATOM 2122 OWO WATW 34 14.542 5.167 15.017 1.0015.88 8 ATOM 2123 OWO WATW 35 15.681 2.641 14.612 1.0016.11 8 ATOM 2124 OWO WATW 36 22.959 -5.924 55.817 1.0016.14 8 ATOM 2125 OWO WATW 37 17.792 -7.760 24.674 1.0016.20 8 ATOM 2126 OWO WATW 38 -5.745 -0.511 32.819 1.0016.38 8 30ATOM 2127 OWO WATW 39 8.911 8.038 13.921 1.0016.56 8 ATOM 2128 OWO WATW 40 0.345 -4.964 25.288 1.0016.65 8 ATOM 2129 OWO WATW 41 11.031 -11.43125.798 1.0016.84 8 ATOM 2130 OWO WATW 42 13.079 5.575 51.582 1.0017.00 8 ATOM 2131 OWO WATW 43 9.891 -5.124 53.560 1.0017.04 8 35ATOM 2132 OWO WATW 44 5.686 -8.148 48.229 1.0017.09 8 ATOM 2133 OWO WATW 45 25.251 13.768 26.180 1.0017.19 8 ATOM 2134 OWO WATW 46 -0.436 1.365 15.687 1.0017.45 8 ATOM 2135 OWO WATW 47 -9.577 -3.555 38.689 1.0017.51 8 ATOM 2136 OWO WATW 48 30.018 9.014 37.735 1.0018.20 8 40ATOM 2137 OWO WATW 49 15.370 -7.155 29.434 1.0018.50 8 ATOM 2138 OWO WATW 50 20.118 20.536 34.111 1.0018.97 8 ATOM 2139 OWO WATW 51 23.269 -5.467 51.272 1.0019.01 8 ATOM 2140 OWO WATW 52 18.707 -18.34845.501 1.0019.39 8 ATOM 2141 OWO WATW 53 29.237 2.519 48.044 1.0019.62 8 45ATOM 2142 OWO WATW 54 -2.442 2.172 47.419 1.0019.63 8 ATOM 2143 OWO WATW 55 19.933 -4.327 22.737 1.0019.76 8 ATOM 2144 OWO WATW 56 34.473 -1.581 37.164 1.0019.96 8 ATOM 2145 OW0 WATW 57 12.821 11.824 47.078 1.0020.13 8 ATOM 2146 OWO WATW 58 11.503 7.499 13.241 1.0020.21 8 50ATOM 2147 OWO WATW 59 23.684 -1.037 20.633 1.0020.22 8 ATOM 2148 OWO WATW 60 -0.612 -3.323 51.692 1.0020.28 8 ATOM 2149 OWO WATW 61 17.007 14.116 44.269 1.0020.65 8 ATOM 2150 OWO WATW 62 -0.273 -9.571 27.817 1.0020.67 8 ATOM 2151 OWO WATW 63 -11.316 -4.375 40.684 1.0020.73 8 55ATOM 2152 OWO WATW 64 25.755 -8.111 33.313 1.0020.89 8 ATOM 2153 4W0 WATW 65 -3.039 -2.283 51.281 1.0020.99 8 ATOM 2154 OWO WATW 66 14.409 -8.631 51.937 1.0020.99 8 ATOM 2155 OWO WATW 67 -3.784 -5.571 30.454 1.0020.99 8 ATOM 2156 OWO WATW 68 28.701 0.916 24.873 1.0021.03 8 60ATOM 2157 OWO WATW 69 -2.298 4.090 45.230 1.0021.11 8 ATOM 2158 OWO WATW 70 30.077 4.491 44.090 1.0021.11 8 ATOM 2159 OWO WATW 71 -4.539 6.907 25.235 1.0021.20 8 ATOM 2160 OW0 WATW 72 7.930 16.602 17.863 1.0021.38 8 ATOM 2161 OWO WATW 73 25.030 5.431 18.766 1.0021.46 8 65ATOM 2162 OWO WATW 74 21.967 -14.31649.321 1.0021.78 8 ATOM 2163 OWO WATW 75 2.376 -6.419 49.074 1.0021.92 8 ATOM 2164 OWO WATW 76 4.085 10.456 48.716 1.0022.11 8 ATOM 2165 OWO WATW 77 5.239 -12.23248.334 1.0022.25 8 ATOM 2166 OWO WATW 78 -4.190 -2.933 30.508 1.0022.26 8 ATOM 2167 OWO WATW 79 8.378 5.587 51.200 1.0022.44 8 ATOM 2168 OWO WATW 80 12.983 -5.518 20.760 1.0022.55 8 ATOM 2169 OWO WATW B1 9.499 10.528 14.890 1.0022.84 B

5 ATOM 2170 OWO WATW 82 2.960 -17.25340.993 1.0023.00 8 ATOM 2171 OWO WATW 83 6.098 -15.84934.785 1.0023.11 8 ATOM 2172 OWO WATW 84 -9.765 -5.816 36.844 1.0023.23 8 ATOM 2173 OWO WATW 85 17.165 -8.431 51.079 1.0023.38 B

ATOM 2174 OWO WATW 86 26.762 4.780 20.872 1.0023.45 8 l0ATOM 2175 OWO WATW 87 -3.582 -0.255 20.689 1.0023.52 8 ATOM 2176 OWO WATW 88 24.998 -0.493 54.746 1.0023.60 8 ATOM 2177 OWO WATW 89 15.378 4.977 52.842 1.0023.62 8 ATOM 2178 OWO WATW 90 -3.290 -9.213 32.314 1.0023.62 8 ATOM 2179 OWO WATW 91 -1.217 9.980 21.173 1.0023.69 8 15ATOM 2180 OWO WATW 92 -4.575 5.139 23.029 1.0023.74 8 ATOM 2181 OWO WATW 93 5.660 -20.27233.874 1.0023.86 8 ATOM 2182 OWO WATW 94 2.570 -19.71732.700 1.0023.93 8 ATOM 2183 OWO WATW 95 -2.768 8.489 18.967 1.0024.13 8 ATOM 2184 OWO WATW 96 -9.884 0.662 45.427 1.0024.32 8 20ATOM 2185 OWO WATW 97 5.619 -4.476 51.362 1.0024.37 8 ATOM 2186 OWO WATW 98 8.421 11.297 38.167 1.0024.65 8 ATOM 2187 OWO WATW 99 25.813 -8.535 52.635 1.0024.70 8 ATOM 2188 OWO WATW 100 20.832 19.605 26.661 1.0024.82 8 ATOM 2189 OWO WATW 101 16.258 -9.256 21.262 1.0024.86 8 25ATOM 2190 OWO WATW 102 12.349 13.826 43.372 1.0024.89 8 ATOM 2191 OWO WATW 103 13.170 -19.74535.451 1.0024.90 8 ATOM 2192 OWO WATW 104 7.075 17.770 20.578 1.0024.93 8 ATOM 2193 OWO WATW 105 22.242 -3.099 22.446 1.0024.94 8 ATOM 2194 OWO WATW 106 2.596 -15.34931.525 1.0025.01 8 30ATOM 2195 OWO WATW 107 13.138 -13.43226.305 1.0025.13 8 ATOM 2196 OWO WATW 108 27.906 13.991 24.255 1.0025.14 8 ATOM 2197 OWO WATW 109 6.218 -4.057 14.703 1.0025.22 8 ATOM 2198 OWO WATW 110 10.505 12.665 32.677 1.0025.29 8 ATOM 2199 OWO WATW 111 -3.781 -2.725 27.641 1.0025.30 8 35ATOM 2200 OWO WATW 112 30.677 10.964 34.167 1.0025.31 8 ATOM 2201 OWO WATW 113 17.661 -13.78150.306 1.0025.32 8 ATOM 2202 OWO WATW 114 34.541 6.057 36.868 1.0025.36 8 ATOM 2203 OWO WATW 115 23.605 3.174 17.711 1.0025.38 8 ATOM 2204 OWO WATW 116 17.497 -13.27824.578 1.0025.43 8 40ATOM 2205 OWO WATW 117 26.337 -11.22548.970 1.0025.54 8 ATOM 2206 OWO WATW 118 -5.239 13.734 29.361 1.0025.59 8 ATOM 2207 OWO WATW 119 -2.765 6.609 16.532 1.0025.61 8 ATOM 2208 OWO WATW 120 -0.782 -2.817 17.108 1.0025.71 8 ATOM 2209 OWO WATW 121 16.158 7.089 14.095 1.0025.77 8 45ATOM 2210 OWO WATW 122 18.930 12.534 48.368 1.0026.12 8 ATOM 2211 OWO WATW 123 24.403 -6.067 53.444 1.0026.65 8 ATOM 2212 OWO WATW 124 -3.404 4.730 49.022 1.0026.81 8 ATOM 2213 OWO WATW 125 32.619 10.296 29.183 1.0026.88 8 ATOM 2214 OW0 WATW 126 -6.804 14.466 42.289 1.0027.19 8 50ATOM 2215 OWO WATW 127 24.517 14.294 40.806 1.0027.26 8 ATOM 2216 OWO WATW 128 -4.697 17.443 41.250 1.0027.26 8 ATOM 2217 OWO WATW 129 15.601 -5.581 15.252 1.0027.49 8 ATOM 2218 OWO WATW 130 19.225 -7.757 52.854 1.0027.55 8 ATOM 2219 OWO WATW 131 20.571 -7.244 23.187 1.0027.79 8 55ATOM 2220 OWO WATW 132 -5.634 12.995 45.863 1.0027.84 8 ATOM 2221 OWO WATW 133 29.455 2.015 28.288 1.0027.85 8 ATOM 2222 OWO WATW 134 35.253 6.005 33.542 1.0027.91 8 ATOM 2223 OWO WATW 135 26.528 7.004 17.380 1.0028.00 8 ATOM 2224 OWO WATW 136 4.802 -2.134 53.088 1.0028.06 8 60ATOM 2225 OWO WATW 137 7.702 -19.31635.292 1.0028.29 8 ATOM 2226 OWO WATW 138 33.637 -3.892 43.427 1.0028.32 8 ATOM 2227 OWO WATW 139 -3.078 -11.20441.616 1.0028.34 8 ATOM 2228 OWO WATW 140 7.296 -11.85520.394 1.0028.39 8 ATOM 2229 OWO WATW 141 -8.355 14.458 38.156 1.0028.47 8 65ATOM 2230 OWO WATW 142 -3.786 -10.07745.809 1.0028.51 8 ATOM 2231 OWO WATW 143 17.884 8.271 55.001 1.0028.52 8 ATOM 2232 OWO WATW 144 -7.450 9.431 27.023 1.0028.66 8 ATOM 2233 OWO WATW 145 25.034 10.848 14.171 1.0028.68 8 ATOM 2234 OWOWAT W146 27.154 14.822 33.2561.00 28.71 B

ATOM 2235 OWOWAT W147 3.930 14.554 35.3531.00 28.86 8 ATOM 2236 OWOWAT W148 3.832 14.101 17.3671.00 28.94 8 ATOM 2237 OWOWAT W149 -7.141 6.522 26.4331.00 28.95 8 ATOM 2238 OWOWAT W150 16.291 15.441 37.7481.00 28.96 B

ATOM 2239 OWOWAT W151 23.732 -13.47232.8131.00 29.06 B

ATOM 2240 OWOWAT W152 31.579 0.528 49.0091.00 29.17 8 ATOM 2241 OWOWAT W153 0.948 11.515 50.8561.00 29.19 8 ATOM 2242 OWOWAT W154 20.562 20.238 24.1041.00 29.61 8 ATOM 2243 OWOWAT W155 14.815 22.549 27.6581.00 29.72 8 ATOM 2244 OWOWAT W156 -0.505 13.461 15.8441.00 29.79 8 ATOM 2245 OWOWAT W157 27.503 -7.381 36.8141.00 29.90 8 ATOM 2246 OWOWAT W158 31.766 -7.236 46.5771.00 29.96 8 ATOM 2247 OWOWAT W159 2.280 5.918 54.2431.00 30.06 8 ATOM 2248 OWOWAT W160 15.109 18.191 36.2481.00 30.13 8 ATOM 2249 OWOWAT W161 4.637 -16.47932.1131.00 30.14 8 ATOM 2250 OWOWAT W162 17.268 13.651 16.6881.00 30.17 8 ATOM 2251 OWOWAT W163 19.452 14.125 43.0371.00 30.18 8 ATOM 2252 OWOWAT W164 -4.171 13.696 26.8861.00 30.24 B

ATOM 2253 OWOWAT W165 14.909 -15.47749.5341.00 30.29 8 ATOM 2254 OWOWAT W166 -8.602 11.318 30.5571.00 30.42 8 ATOM 2255 OWOWAT W167 19.207 -15.05828.1591.00 30.52 8 ATOM 2256 OWOWAT W168 26.601 10.511 46.9691.00 30.58 8 ATOM 2257 OWOWAT W169 31.110 -8.170 41.3591.00 30.61 8 ATOM 2258 OWOWAT W170 29.593 8.135 46.3491.00 30.72 8 ATOM 2259 OWOWAT W171 -10.368 -1.876 34.5041.00 30.74 8 ATOM 2260 OWOWAT W172 28.564 -4.100 29.5441.00 30.83 B

ATOM 2261 OWOWAT W173 -12.777 4.044 45.4101.00 30.92 B

ATOM 2262 OWOWAT W174 7.794 -21.93142.3191.00 30.96 8 ATOM 2263 OWOWAT W175 18.808 -10.25123.6881.00 31.07 8 ATOM 2264 OWOWAT W176 0.113 -6.364 50.9141.00 31.09 8 ATOM 2265 OWOWAT W177 -3.585 3.671 16.7851.00 31.12 8 ATOM 2266 OWOWAT W178 4.754 -21.90138.8261.00 31.24 8 ATOM 2267 OWOWAT W179 3.124 -4.459 52.0131.00 31.30 8 ATOM 2268 OWOWAT W180 27.364 15.293 27.0981.00 31.43 8 ATOM 2269 OWOWAT W181 19.204 -18.62042.6331.00 31.46 8 ATOM 2270 OWOWAT W182 23.808 -11.49540.0591.00 31.53 8 ATOM 2271 OWOWAT W183 29.332 -1.923 29.9531.00 31.57 8 ATOM 2272 OWOWAT W184 12.448 14.328 33.0701.00 31.59 8 ATOM 2273 OWOWAT W185 1.205 17.345 29.9811.00 31.67 8 ATOM 2274 OWOWAT W186 -9.791 7.997 34.8441.00 31.75 8 ATOM 2275 OWOWAT W187 -7.837 18.408 38.0691.00 31.89 8 ATOM 2276 OWOWAT W188 11.140 -9.008 50.7921.00 31.95 8 ATOM 2277 OWOWAT W189 26.511 -2.526 54.7601.00 32.13 8 ATOM 2278 OWOWAT W190 23.093 -7.348 24.1921.00 32.27 8 ATOM 2279 OWOWAT W191 -10.284 6.288 39.3791.00 32.43 8 ATOM 2280 OWOWAT W192 -7.821 -0.312 31.3581.00 32.44 8 ATOM 2281 OWOWAT W193 20.703 -19.05840.1281.00 32.50 8 ATOM 2282 OWOWAT W194 23.085 18.180 25.2981.00 32.52 8 ATOM 2283 OWOWAT W195 18.564 11.924 14.8831.00 32.61 8 ATOM 2284 OWOWAT W196 19.725 -15.77637.2271.00 32.93 8 ATOM 2285 OWOWAT W19'1 9.423 -12.85050.0291.00 33.07 8 ATOM 2286 OWOWAT W198 -5.226 -11.89139.0401.00 33.31 8 ATOM 2287 OWOWAT W199 -10.872 11.311 41.6221.00 33.34 8 ATOM 2288 OWOWAT W200 24.953 -10.12351.1081.00 33.47 8 ATOM 2289 OWOWAT W201 10.234 12.343 37.4421.00 33.61 8 ATOM 2290 OWOWAT W202 -1.385 9.325 49.5901.00 33.68 8 ATOM 2291 OWOWAT W203 13.133 -13.56250.5161.00 33.68 8 ATOM 2292 OWOWAT W204 32.332 3.720 31.2301.00 33.72 8 ATOM 2293 OWOWAT W205 -4.769 19.603 30.8901.00 34.01 8 ATOM 2294 OWOWAT W206 -10.676 2.037 32.3731.00 34.14 8 ATOM 2295 OWOWAT W207 5.473 -14.54147.4181.00 34.18 8 ATOM 2296 OWOWAT W208 -0.600 -4.653 18.9591.00 34.35 8 ATOM 2297 OWOWAT W209 5.122 13.867 48.9791.00 34.37 8 ATOM 2298 OWOWAT W210 -4.776 -9.796 38.6961.00 34.40 8 ATOM 2299 OWOWAT W211 22.711 8.507 56.1511.00 34.54 8 ATOM 2300 OWOWAT W212 -5.723 12.192 25.1991.00 34.59 8 ATOM 2301 OWOWAT W213 -5.854 7.368 47.0361.00 34.60 8 ATOM 2302 OWOWAT W214 2.162 12.775 15.4721.00 34.69 8 ATOM 2303 OWOWAT W215 29.086 -4.835 51.2441.00 34.91 8 ATOM 2304 OWOWAT W216 29.521 1.500 30.2901.00 35.03 8 ATOM 2305 OWOWAT W217 9.270 16.229 27.6471.00 35.08 8 ATOM 2306 OWOWAT W218 -0.559 -13.99044.9421.00 35.09 8 ATOM 2307 OWOWAT W219 31.092 12.772 28.1021.00 35.13 8 ATOM 2308 OWOWAT W220 4.053 17.330 40.6491.00 35.18 8 ATOM 2309 OWOWAT W221 9.804 12.126 12.8061.00 35.19 8 ATOM 2310 OWOWAT W222 16.382 10.037 14.0841.00 35.33 8 10ATOM 2311 OWOWAT W223 34.860 8.861 43.0501.00 35.36 8 ATOM 2312 OWOWAT W224 2.481 -1.469 55.1851.00 35.39 8 ATOM 2313 OWOWAT W225 27.639 15.901 20.2201.00 35.45 8 ATOM 2314 OWOWAT W226 13.522 14.546 22.1931.00 35.58 8 ATOM 2315 OWOWAT W227 18.759 -16.36834.3411.00 35.64 8 15ATOM 2316 OWOWAT W228 29.746 6.054 47.9831.00 35.88 8 ATOM 2317 OWOWAT W229 1.824 8.703 50.4411.00 35.91 8 ATOM 2318 OWOWAT W230 4.304 -10.21220.5661.00 36.11 8 ATOM 2319 OWOWAT W231 25.903 -4.307 53.0391.00 36.25 8 ATOM 2320 OWOWAT W232 30.041 -9.858 50.3141.00 36.32 8 20ATOM 2321 OWOWAT W233 2.098 9.375 12.7241.00 36.32 8 ATOM 2322 OWOWAT W234 -6.517 -10.58746.8461.00 36.56 8 ATOM 2323 OWOWAT W235 -6.610 -3.836 30.4151.00 36.57 8 ATOM 2324 OWOWAT W236 -10.495 12.363 34.8991.00 36.64 8 ATOM 2325 OWOWAT W237 -9.368 9.062 33.0121.00 36.76 8 25ATOM 2326 OWOWAT W238 19.878 23.075 33.2881.00 36.92 8 ATOM 2327 OWOWAT W239 -4.530 7.046 20.8961.00 36.93 8 ATOM 2328 OWOWAT W240 33.313 6.152 46.2021.00 36.93 B

ATOM 2329 OWOWAT W241 -8.607 4.039 46.9241.00 37.16 8 ATOM 2330 OWOWAT W242 -0.158 -8.511 20.7281.00 37.69 8 30ATOM 2331 OWOWAT W243 5.833 13.274 13.5961.00 37.75 8 ATOM 2332 OWOWAT W244 5.857 -19.50331.1981.00 37.77 8 ATOM 2333 OWOWAT W245 -2.468 -11.12530.4961.00 37.88 8 ATOM 2334 OWOWAT W246 8.010 -18.25025.5541.00 37.97 8 ATOM 2335 OWOWAT W247 -2.981 10.860 22.6071.00 38.01 B

35ATOM 2336 OWOWAT W248 29.733 2.478 51.1851.00 38.06 8 ATOM 2337 OWOWAT W249 -1.876 18.713 35.6921.00 38.18 8 ATOM 2338 OWOWAT W250 -0.040 -2.395 54.3651.00 38.20 8 ATOM 2339 OWOWAT W251 -2.499 -1.254 18.1431.00 38.26 8 ATOM 2340 OWOWAT W252 1.301 15.936 18.0641.00 38.65 8 40ATOM 2341 OWOWAT W253 -7.703 5.024 28.8411.00 38.66 8 ATOM 2342 OWOWAT W254 8.197 -10.54851.1051.00 38.97 8 ATOM 2343 OWOWAT W255 19.072 -5.777 16.6001.00 39.02 8 ATOM 2344 OWOWAT W256 -1.755 -6.479 25.7041.00 39.11 8 ATOM 2345 OWOWAT W257 15.948 -20.84638.3421.00 39.37 8 45ATOM 2346 OWOWAT W258 -7.884 13.866 29.1481.00 39.59 8 ATOM 2347 OWOWAT W259 34.511 11.821 32.7231.00 39.65 8 ATOM 2348 OWOWAT W260 16.479 -16.08427.9521.00 39.69 8 ATOM 2349 OWOWAT W261 -8.601 2.060 30.4561.00 39.87 8 ATOM 2350 OWOWAT W262 -0.861 17.301 21.8491.00 39.89 8 50ATOM 2351 OWOWAT W263 8.555 -18.27547.2411.00 39.93 8 ATOM 2352 OWOWAT W264 24.230 -5.252 22.6641.00 40.00 8 ATOM 2353 OWOWAT W265 -1.056 0.937 53.9221.00 40.53 8 ATOM 2354 OWOWAT W266 16.017 -13.90222.3261.00 40.63 8 ATOM 2355 OWOWAT W267 23.066 10.127 50.3341.00 40.86 8 55ATOM 2356 OWOWAT W268 12.877 15.614 35.0231.00 40.87 8 ATOM 2357 OWOWAT W269 21.711 -4.797 18.7611.00 40.90 8 ATOM 2358 OWOWAT W270 28.676 -7.905 40.7391.00 41.51 8 ATOM 2359 OWOWAT W271 21.557 -6.991 52.2771.00 42.05 8 ATOM 2360 OWOWAT W272 18.619 5.353 14.6611.00 42.32 8 60ATOM 2361 OWOWAT W273 6.542 -6.740 51.8521.00 42.53 8 ATOM 2362 OWOWAT W274 13.730 15.335 37.5371.00 42.69 8 ATOM 2363 OWOWAT W275 25.430 5.894 14.8161.00 42.71 8 ATOM 2364 OWOWAT W276 -6.269 3.726 22.2881.00 43.87 8 ATOM 2365 OWOWAT W277 19.099 -16.34931.9121.00 43.95 8 65ATOM 2366 OWOWAT W278 19.470 8.026 13.8181.00 43.97 8 ATOM 2367 OWOWAT W279 22.549 19.383 22.0281.00 44.26 8 ATOM 2368 OWOWAT W280 -7.882 -11.62439.5781.00 44.88 8 ATOM 2369 OWOWAT W281 12.425 -9.624 21.3921.00 45.09 8 WO 00/22103 ~PCT/DK99/00542 ATOM 2370 OWO WATW 282 9.040 -7.99613.289 1.0045.24 ATOM 2371 OWO WATW 283 18.170 -7.82217.373 1.0045.27 ATOM 2372 OWO WATW 284 20.862 6.192 13.601 1.0045.89 ATOM 2373 OWO WATW 285 7.780 -19.94130.094 1.0046.04 ATOM 2374 OWO WATW 286 25.580 16.28635.358 1.0046.89 ATOM 2375 OWO WATW 287 16.268 22.91235.142 1.0047.83 ATOM 2376 OWO WATW 288 7.741 15.09227.401 1.0048.86 ATOM 2377 OWO WATW 289 30.772 12.83522.683 1.0049.34 ATOM 2378 OWO WATW 290 22.334 12.13249.136 1.0049.76 ATOM2379 OWO WATW 291 -9.173 -1.10347.956 1.0050.16 WO 00/22103 .PCT/DK99/00542 SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i)APPLICANT:

(A) NAME: Novo Nordisk A/S

(B) STREET: Novo Alle (C) CITY: Bagsveard (E) COUNTRY: Denmark (F) POSTAL CODE (ZIP):

(G) TELEPHONE: +45 4444 (H) TELEFAX: +45 4449 3256 (ii)TITLE OF INVENTION: A modifiedpolypeptide (iii) NUMBER
OF
SEQUENCES:

(iv) COMPUTER
READABLE
FORM:

(A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: PatentIn 0, Release #1. Version #1.30 (EPO) (2)INFORMATION
FOR
SEQ
ID
NO:
1:

(i)SEQUENCE CHARACTERISTICS:

(A) LENGTH: 840 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii)MOLECULE TYPE: DNA (genomic) (vi)ORIGINAL SOURCE:

(B) STRAIN: Bacillus sp. No. 40484 PD498, NCIMB

(ix)FEATURE:

(A) NAME/KEY: CDS

(B) LOCATION:1..840 (xi)SEQUENCE DESCRIPTION: SEQ
ID NO: 1:

TCT GCT

TrpSerPro Asn Asp Pro Tyr Tyr TyrGlnTyr Gly Pro Gln Ser Ala GAT GTA

AsnThrSer Thr Pro Ala Ala Trp ThrArgGly Ser Ser Thr Asp Val GGA GTG

GlnThrVal Ala Val Leu Asp Ser AspTyr.AsnHis Pro Asp Gly Val TAC GAC

LeuAlaArg Lys Val Ile Lys Gly PheIle.AspArg Asp Asn Tyr Asp GGT ACC

AsnProMet Asp Leu Asn Gly His HisValAla Gly Thr Val Gly Thr GGC GTA

AlaAlaAsp Thr Asn Asn Gl.y Ile AlaGlyMet Ala Pro Asp Gly Val CTT GAT

ThrLysIle Leu Ala Val Arg Val AlaAsnGly Ser Gly Ser Leu Asp 100 105 i10 Leu Asp Ser Ile Ala Ser Gly Ile Arg Tyr Ala Ala Asp Gln Gly Ala Lys Val Leu Asn Leu Ser Leu Gly Cys Glu Cys Asn Ser Thr Thr Leu Lys Ser Ala Val Asp Tyr Ala Trp Asn Lys Gly Ala Val Val Val Ala Ala Ala Gly Asn Asp Asn Val Ser Arg Thr Phe Gln Pro Ala Ser Tyr Pro Asn Ala Ile Ala Val Gly Ala Ile Asp Ser Asn Asp Arg Lys Ala AAT TAC ACG GTG
GAT
GTC
ACT
GCT

SerPhe Ser Tyr Gly TrpVal Asp Val Thr ProGly Val Asn Thr Ala TCA CCG TAC

AsnIle Ala Thr Val AsnAsn Gly Tyr Ser MetSer Gly Ser Pro Tyr GCA CAC GCT

ThrSer Met Ser Pro ValAla Gly Leu Ala LeuLeu Ala Ala His .Ala AAG GTA ATT

SerGln Gly Asn Asn GlnIle Arg Gln Ala GluGln Thr Lys Val Ile ATC ACT TAT

AlaAsp Lys Ser Gly GlyThr Asn Phe Lys GlyLys Ile Ile Thr Tyr Asn Ser Asn Lys Ala Val Arg Tyr (2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 280 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2:
Trp Ser Pro Asn Asp Pro Tyr Tyr Ser Ala Tyr Gln Tyr Gly Pro Gln Asn Thr Ser Thr Pro Ala Ala Trp Asp Val Thr Arg Gly Ser Ser Thr Gln Thr Val Ala Val Leu Asp Ser Gly Val Asp Tyr Asn His Pro Asp Leu Ala Arg Lys Val Ile Lys Gly Tyr Asp Phe Ile Asp Arg Asp Asn Asn Pro Met Asp Leu Asn Gly His Gly Thr His Val Ala Gly Thr Val Ala Ala Asp Thr Asn Asn Gly Ile Gly Val Ala Gly Met Ala Pro Asp Thr Lys Ile Leu Ala Val Arg Val Leu Asp Ala Asn Gly Ser Gly Ser Leu Asp Ser Ile Ala Ser Gly Ile Arg Tyr Ala Ala Asp Gln Gly Ala Lys Val Leu Asn Leu Ser Leu Gly Cys Glu Cys Asn Ser Thr Thr Leu Lys Ser Ala Val Asp Tyr Ala Trp Asn Lys Gly Ala Val Val Val Ala Ala Ala Gly Asn Asp Asn Val Ser Arg Thr Phe Gln Pro Ala Ser Tyr Pro Asn Ala Ile Ala Val Gly Ala Ile Asp Ser Asn Asp Arg Lys Ala Ser Phe Ser Asn Tyr Gly Thr Trp Val Asp Val Thr Ala Pro Gly Val Asn Ile Ala Ser Thr Val Pro Asn Asn Gly Tyr Ser Tyr Met Ser Gly Thr Ser Met Ala Ser Pro His Val Ala Gly Leu Ala Ala Leu Leu Ala Ser Gln Gly Lys Asn Asn Val Gln Ile Arg Gln Ala Ile Glu Gln Thr Ala Asp Lys Ile Ser Gly Thr Gly Thr Asn Phe Lys Tyr Gly Lys Ile Asn Ser Asn Lys Ala Val Arg Tyr (2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 269 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (vi) ORIGINAL SOURCE:
(B) STRAIN: Bacillus lentus (xi) SEQUENCE DESCRIPTTON: SEQ ID NO: 3:
Ala Gln Ser Val Pro Trp Gly Ile Ser Arg Val Gln Ala Pro Ala Ala His Asn Arg Gly Leu Thr Gly Ser Gly Val Lys Val Ala Val Leu Asp Thr Gly Ile Ser Thr His Pro Asp Leu Asn Ile Arg Gly Gly Ala Ser Phe Val Pro Gly Glu Pro Ser Thr Gln Asp Gly Asn Gly His Gly Thr His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu Gly Val Ala Pro Ser Ala Glu Leu Tyr Ala Val Lys Val Leu Gly Ala Ser Gly Ser Gly Ser Val Ser Ser Ile Ala Gln Gly Leu Glu Trp Ala Gly Asn Asn Gly Met His Val Ala Asn Leu Ser Leu Gly Ser Pro Ser Pro Ser Ala Thr Leu Glu Gln Ala Val Asn Ser Ala Thr Ser Arg Gly Val Leu Val Val Ala Ala Ser Gly Asn Ser Gly Ala Gly Ser Ile Ser Tyr Pro Ala Arg Tyr Ala Asn Ala Met Ala Val Gly Ala Thr Asp Gln Asn Asn Asn Arg Ala Ser Phe Ser Gln Tyr Gly Ala Gly Leu Asp Ile Val Ala Pro Gly Val Asn Val Gln Ser Thr Tyr Pro Gly Ser Thr Tyr Ala Ser Leu Asn Gly Thr Ser Met Ala Thr Pro His Val Ala Gly Ala Ala Ala Leu Val Lys Gln Lys Asn Pro Ser Trp Ser Asn Val Gln Ile Arg Asn His Leu Lys Asn Thr Ala Thr Ser Leu Gly Ser Thr Asn Leu Tyr Gly Ser Gly Leu Val Asn Ala Glu Ala Ala Thr Arg (2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 1458 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (vi) ORIGINAL SOURCE:

(B) STRAIN: Bacillus sp. DSM No. 12649 (ix) FEATURE:
(A) NAME/KEY: CDS
(B) LOCATION:1..1458 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4:
cac cat aat ggt acg aac ggc aca atg atg cag tac ttt gaa tgg tat 48 His His Asn Gly Thr Asn Gly Thr Met Met Gln Tyr Phe Glu Trp Tyr cta cca aat gac gga aac cat tgg aat aga tta agg tct gat gca agt 96 Leu Pro Asn Asp Gly Asn His Trp Asn Arg Leu Arg Ser Asp Ala Ser aac cta aaa gat aaa ggg atc tca gcg gtt tgg att cct cct gca tgg 144 Asn Leu Lys Asp Lye Gly Ile Ser Ala Val Trp Ile Pro Pro Ala Trp aag ggt gcc tct caa aat gat gtg ggg tat ggt get tat gat ctg tat 192 Lys Gly Ala Ser Gln Asn Asp Val Gly Tyr Gly Ala Tyr Asp Leu Tyr gat tta gga gaa ttc aat caa aaa gga acc att cgt aca aaa tat gga 240 Asp Leu Gly Glu Phe Asn Gln Lys Gly Thr Ile Arg Thr Lys Tyr Gly acg cgc aat cag tta caa get gca gtt aac gcc ttg aaa agt aat gga 288 Thr Arg Asn Gln Leu Gln Ala Ala Val Asn Ala Leu Lys Ser Asn Gly att caa gtg tat ggc gat gtt gta atg aat cat aaa ggg gga gca gac 336 Ile Gln Val Tyr Gly Asp Val Val Met Asn His Lys Gly Gly Ala Asp get acc gaa atg gtt agg gca gtt gaa gta aac ccg aat aat aga aat 384 Ala Thr Glu Met Val Arg Ala Val Glu Val Asn Pro Asn Asn Arg Asn caa gaa gtg tcc ggt gaa tat aca att gag get tgg aca aag ttt gac 432 Gln Glu Val Ser Gly Glu Tyr Thr Ile Glu Ala Trp Thr Lys Phe Asp ttt cca gga cga ggt aat act cat tca aac ttc aaa tgg aga tgg tat 480 Phe Pro Gly Arg Gly Asn Thr His Ser Asn Phe Lys Trp Arg Trp Tyr cac ttt gat gga gta gat tgg gat cag tca cgt aag ctg aac aat cga 528 His Phe Asp Gly Val Asp Trp Asp Gln Ser Arg Lys Leu Asn Asn Arg att tat aaa ttt aga ggt gat gga aaa ggg tgg gat tgg gaa gtc gat 576 Ile Tyr Lys Phe Arg Gly Asp Gly Lys Gly Trp Asp Trp Glu Val Asp aca gaa aac ggt aac tat gat tac cta atg tat gca gat att gac atg 624 Thr Glu Asn Gly Asn Tyr Asp Tyr Leu Met Tyr Ala Asp Ile Asp Met gat cac cca gag gta gtg aat gag cta aga aat tgg ggt gtt tgg tat 672 Asp His Pro Glu Val Val Asn Glu Leu Arg Asn Trp Gly Val Trp Tyr acg aat aca tta ggc ctt gat ggt ttt aga ata gat gca gta aaa cat 720 Thr Asn Thr Leu Gly Leu Asp Gly Phe Arg Ile Asp Ala Val Lys His ata aaa tac agc ttt act cgt gat tgg att aat cat gtt aga agt gca 768 Ile Lys Tyr Ser Phe Thr Arg Asp Trp Ile Asn His Val Arg Ser Ala act ggc aaa aat atg ttt gcg gtt gcg gaa ttt tgg aaa aat gat tta 816 Thr Gly Lys Asn Met Phe Ala Val Ala Glu Phe Trp Lys Asn Asp Leu ggt get att gaa aac tat tta aac aaa aca aac tgg aac cat tca gtc 864 Gly Ala Ile Glu Asn Tyr Leu Asn Lys Thr Asn Trp Asn His Ser Val ttt gat gtt ccg ctg cac tat aac ctc tat aat get tca aaa agc gga 912 Phe Asp Val Pro Leu His Tyr Asn Leu Tyr Asn Ala Ser Lys Ser Gly ggg aat tat gat atg agg caa ata ttt aat ggt aca gtc gtg caa aga 960 Gly Asn Tyr Asp Met Arg Gln Ile Phe Asn Gly Thr Val Val Gln Arg cat cca atg cat get gtt aca ttt gtt gat aat cat gat tcg caa cct 1008 His Pro Met His Ala Val Thr Phe Val Asp Asn His Asp Ser Gln Pro gaa gaa get tta gag tct ttt gtt gaa gaa tgg ttc aaa cca tta gcg 1056 Glu Glu Ala Leu Glu Ser Phe Val Glu Glu Trp Phe Lys Pro Leu Ala tat get ttg aca tta aca cgt gaa caa ggc tac cct tct gta ttt tat 1104 Tyr Ala Leu Thr Leu Thr Arg Glu Gln Gly Tyr Pro Ser Val Phe Tyr gga gat tat tat ggc att cca acg cat ggt gta cca gcg atg aaa tcg 1152 Gly Asp Tyr Tyr Gly Ile Pro Thr His Gly Val Pro Ala Met Lys Ser aaa att gac ccg att cta gaa gcg cgt caa aag tat gca tat gga aga 1200 Lys Ile Asp Pro Ile Leu Glu Ala Arg Gln Lys Tyr Ala Tyr Gly Arg caa aat gac tac tta gac cat cat aat atc atc ggt tgg aca cgt gaa 1248 Gln Asn Asp Tyr Leu Asp His His Asn Tle Ile Gly Trp Thr Arg Glu ggg aat aca gca cac ccc aac tcc ggt tta get act atc atg tcc gat 1296 Gly Asn Thr Ala His Pro Asn Ser Gly Leu Ala Thr Ile Met Ser Asp ggg gca gga gga aat aag tgg atg ttt gtt ggg cgt aat aaa get ggt 1344 Gly Ala Gly Gly Asn Lys Trp Met Phe Val Gly Arg Asn Lys Ala Gly caa gtt tgg acc gat atc act gga aat cgt gca ggt act gtt acg att 1392 Gln Val Trp Thr Asp Ile Thr Gly Asn Arg Ala Gly Thr Val Thr Ile aat get gat gga tgg ggt aat ttt tct gta aat gga gga tca gtt tct 1440 Asn Ala Asp Gly Trp Gly Asn Phe Ser Val Asn Gly Gly Ser Val Ser att tgg gta aac aaa taa 1458 Ile Trp Val Asn Lye

Claims (34)

Claims
1. A polypeptide with reduced immune response, having one or more amino acid residues modified, wherein the C .alpha.-atoms of said amino acid residues are located less than 15 .ANG. from a ligand bound to said polypeptide.
2. A polypeptide according to claim 1, wherein the polypeptide has reduced allergenicity.
3. The polypeptide according to any of the claims 1 to 2, wherein the Ca-atom of the amino acid residues is located closer to the ligand than the C .alpha.-atom.
4. The polypeptide according to any of the claims 1 to 3, wherein the C .alpha.-atoms of the amino acid residues are located less than 10 .ANG. from the ligand and said amino acid residues have an accessibility of at least 15%.
5. The polypeptide according to any of the claims 1 to 4, wherein the ligand is a metal or metal ion.
6. The polypeptide according to any of the preceding claims, wherein the polypeptide is modified by substitution of amino acid residues.
7. The polypeptide according to any of the claims 1 to 6, wherein the modified polypeptide has been selected from a diverse library of variants.
8. The polypeptide according to claim 6, wherein the substituting amino acids contain amino groups in the form of Lysine residues(s), or carboxylic groups in the form of Aspartic acid or Glutamic acid residues, or SH-groups in the form of Cysteine residues.
9. The polypeptide according to claim 6, wherein the modifications) is(are) prepared by a conservative substitution of an amino acid residue, such as an Arginine to Lysine substitution or Aspargine to Aspartate/Glutamate or a Glutamine to Aspartate/Glutamate substitution or Threonine/Serine to Cysteine.
10. The polypeptide according to claims 1-9, wherein the polypeptide is modified by coupling one or more polymeric molecules to said polypeptide, thereby providing a polypeptide-polymer conjugate.
11. The polypeptide according to claim 10, wherein the parent polypeptide moiety of the conjugate has a molecular weight from 1 to 1000 kDa, preferred 4 to 100 kDa, more preferred 12 to 60 kDa.
12. The polypeptide according to claim 10, wherein the polymeric molecules coupled to the polypeptide have a molecular weight from 0.1 to 100, preferably 0.1 to 60 kDa, more preferably 0.3-5 kDa, most preferably 1 to 2 kDa.
13. The polypeptide according to any of the preceding claims, wherein said polypeptide or parent polypeptide is an enzyme selected from the group of Oxidoreductases, including laccases and Superoxide dismutase (SOD); Hydrolases, including carbohydrases, amylases, proteases, especially subtilisins;
Transferases, including Transglutaminases (TGases); Isomerases, including Protein disulfide Isomerases (PDT); Lyases, including Pectate lyases.
14. The polypeptide according to claim 13, wherein said polypeptide or parent polypeptide is PD498, Savinase~, BPN', Amylase, Proteinase K, Proteinase R, Subtilisin DY, Lion Y, Rennilase~, JA16, Alcalase~.
15. The polypeptide according to claim 14, wherein the polypeptide or parent polypeptide of the conjugate is a PD498 variant with one or more of the following substitutions: The amino acid residues in position 86, 87, 7, 47, 51, 219, 12, 218, 10, 11, 53, 28, 1, 65, 61, 63, 67, 60, 69, 55, 44, 45, 111, 115, 109, 215, 200, 202, 170, 268, 250, 152, 254, 136, 269, 246, 141 is substituted with K, D, E, or C, preferably R250K, R250D, R250E, R250C.
16. The polypeptide according to claim 14, wherein the polypeptide or parent polypeptide is a BPN' variant with one or more of the following substitutions: The amino acid residues in position 77, 2, 5, 43, 214, 206, 22, 215, 14, 17, 9, 36, 211, 195, 197, 154, 163, 247, 265, 251, 143, 127, 260, 131, 128, 243 is substituted with K, D, E, or C, preferably R247K, R247D, R247E, R247C.
17. The polypeptide according to claim 14, wherein the polypeptide or parent polypeptide is a Savinase~ variant with one or more of the following substitutions: The amino acid residues in position 75, 2, 42, 208, 200, 14, 22, 17, 189, 241, 125, 125, 141, 245, 259, 237, 254, 157 is substituted with K, D, E, or C, preferably R241K, R241D, R241E, R241C.
18. The polypeptide according to claim 14, wherein the polypeptide or parent polypeptide is an amylase variant with one or more of the following substitutions: The amino acid residue in position 124, 126, 128, 159, 160, 166, 185, 186, 189, 190, 193, 194, 195, 196, 198, 201, 202, 203, 209, 210, 214, 242, 244, 247, 296, 298, 299, 302, 303, 304, 306, 307, 308, 310, 311, 314, 345, 347, 405, 406, 407, 408, 409, 433, 434, 435, 436, 437, 475, 476, 477, 478 is substituted with K, D, E, or C.
19. The polypeptide according to claims 10 or 12, wherein the polymeric molecule is selected from the group comprising a natural or synthetic homo- and heteropolymers, selected from the group of the synthetic polymeric molecules including Branched PEGS, poly-vinyl alcohol (PVA), poly-carboxyl acids, poly-(vinylpyrolidone) and poly-D,L-amino acids, or natural occurring polymeric molecules including dextrans, including carboxymethyl-dextrans, and celluloses such as methylcellulose, carboxymethylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, and hydrolysates of chitosan, starches, such as hydroxyethyl-starches, hydroxypropyl-starches, glycogen, agarose, guar gum, inulin, pullulans, xanthan gums, carrageenin, pectin and alginic acid.
20. The polypeptide according to claim 19, wherein the modified polypeptide is savinase variant R241KbPEG1000 or R241KbPEG2000.
21. The polypeptide according to any of claims 1 to 7, wherein the modified polypeptide is savinase variants R241Q, R241E, R241H or R241K.
22. A method for preparing polypeptides with reduced immune response comprising the steps of:
a) identifying amino acid residues located on the surface of the 3-dimensional structure of the parent polypeptide in question, b) selecting target amino acid residues on the surface of said 3-dimensional structure of said parent polypeptide to be modified, c) substituting one or more amino acid residues selected in step b) with other amino acid residues, and/or d) coupling polymeric molecules to the amino acid residues in step b)and/or step c).
23. The method according to claim 22, wherein the C .alpha.-atoms of the amino acid residues are located less than 15 .ANG. from the ligand bound to said polypeptide.
24. The method according to any of claims 22 to 23, wherein the C .beta.-atoms of the amino acid residues are located closer to the ligand than the C .alpha.-atom.
25. The method according to any of the claims 22-24, wherein the C .alpha.-atoms of the amino acid residues are located less than 10 .ANG. from the ligand and said amino acid residues have an accessibility of at least 15%, preferable at least 20%, more preferably at least 30%.
26. The method according to any of the claims 22-25, wherein the identification of amino acid residues located on the surface on the polypeptide referred to in step a) are performed by a computer program analyzing the 3-dimensional structure of the parent polypeptide in question.
27. The method according to any of the claims 22 to 26, wherein step b) comprises selecting Arginine or Lysine residues on the surface of the parent polypeptide.
28. The method according to claim 27, wherein one or more Arginine residues identified in step b) is(are) substituted with a Lysine residues) in step c).
29. Use of the modified polypeptide in claims 1 to 21 for reducing the allergenicity of industrial products.
30. Use of the modified polypeptide in claims 1 to 21 for reducing the immunogenicity of pharmaceuticals.
31. A composition comprising a modified polypeptide of any of claims 1 to 21 and further comprising ingredients used in industrial products.
32. The composition according to claim 31, wherein the industrial product is a detergent, such as a laundry, dish wash or hard surface cleaning product, including bio-film products or a food or feed product or a textile product.
33. The composition according to claim 32, comprising a modified polypeptide of any of claims 1 to 21 and further ingredients used in personal care products, especially skin care products.
34. A composition comprising a modified polypeptide of any of claims 1 to 21 and further comprising ingredients used in pharmaceuticals.
CA002346929A 1998-10-13 1999-10-12 A modified polypeptide with reduced immune response Abandoned CA2346929A1 (en)

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