SYNTHETIC PROTEINS AS IMPLANTABLES

INTRODUCTION

Technical Field

The field of this invention is the production and use of bioresorbable polypeptide polymers.

Background The rate at which an implanted material resorbs or biodegrades within the body can be a major factor in determining its utility as a biomaterial. So called inert materials, such as metals, ceramics and plastics have been shown to be useful for permanent implants. However, in applications in which a device serves as an aid to healing or as a temporary aid in surgical repair, a resorbable material has the advantage of not having to be removed, once healing has occurred. Resorbable sutures and staples, bone pins and screws, wound dressings, and injectable drug delivery systems or depots are examples of such devices. There are very few materials available today which have the physical, chemical and biological properties necessary for the fabrication of medical devices, which must degrade and resorb in the body without detrimental consequences. Various synthetic organic polymers have found use, such as polylactides, polyglycolides, polyanhydrides and polyorthoesters, which degrade in the body by hydrolysis. Collagen, glycosaminoglycans and hyaluronic acid are examples of natural implantable materials which resorb at least partially by enzymatic degradation. The rates of

resorption are limited to the nature of the particular material and modifications can change the rate of resorption, but at the same time may adversely affect the desired properties of the product. Illustrative of efforts to vary resorption characteristics by compositional changes are synthetic resorbable sutures composed of copolymers of lactide and glycolide. By varying the ratio of lactic acid to glycolic acid, the rate of resorption may be varied. Unfortunately, rapidly resorbing compositions tend to be soft and weak. Slow resorbing compositions are stiff and strong. However, their resorption, which is hydrolytic, produces acid buffered by the tissue medium, where erosion occurs at the polymer surface. In addition, however, hydrolysis may occur internally, where the resulting acid catalyzes and accelerates the degradation of the polymer. Thus, internal pockets of degradation can lead to rapid and catastrophic failure of mechanical properties.

There is, therefore, a need for products which can be used in the production of implantable devices. Such products should have the desired mechanical properties of tensile strength, elasticity, formability, and the like, provide for controlled resorption, and be physiologically acceptable.

Relevant Literature

U.S. Patent No. 5,243,038 describes the preparation of high molecular weight, protein polymers and copolymers comprising long segments of small repeating units. Bioactive Polymeric Systems, Gebelein, C. G. and Carraher, C. E., eds., Plenum Press, New York, 1985; Contemporary Biomaterials, Boretos, John . and Eden, Murray, eds., Noyes Publications, New Jersey, 1984; and Concise Guide to Biomedical Polymers: Their Design, Fabrication and Molding, Boretos, John W. , Thomas pub., Illinois, 1973, describe compositions, characteristics, and applications of biomaterials.

SUMMARY OF THE INVENTION

Protein copolymers are provided having segments varying in the number of repetitive units, based on fibroin and elastin. The protein copolymers and silk homopolymers find use in the production of a wide variety of implantable devices and components thereof.

DESCRIPTION OF SPECIFIC EMBODIMENTS Implantable devices and components thereof are provided comprised of recombinant novel copolymers having alternating segments of repetitive units based on fibroin (silk) in combination with elastin or recombinant polymers of fibroin. Particularly, the units for the most part are GAGAGS (SEQ ID NO:01) and VPGVG (SEQ ID NO:02) , respectively, although some variations are permitted, such as the particular order of the amino acids in the sequence and conservative substitutions, such as, but not limited to, replacing serine with threonine and glycine with alanine.

In the copolymers, by varying the ratio of the two different units, the length of the segments comprising each of the units, the molecular weight, any intervening sequences, modifications to the individual repeating units, and the like, one can vary the tensile properties of the product only moderately, such as elasticity, stiffness, hardness, ease of processing, and flexibility, while one can substantially vary the rate of resorption. Faster resorption can be achieved by . reducing the number of repeating units of silk in the silk segment below about 8 units or increasing the number of elastin units per elastin segment to greater than 8, individually or in combination.

For the copolymers, the ratio of the average number of elastin units to the average number of silk units per segment of the repetitive units will be in the range of about 0.5, usually about 1-5. For the most part, there will be at least two fibroin units per segment and not more than about 12, usually not more than about ten, preferably ranging from about 2-8. For the elastin units, there will

usually be at least two, more usually at least about four, generally ranging from about 6-32, more usually from about 6-18, preferably from about 6-16. The percent of amino acids contributed by the silk units will generally range from about 15-65%, more usually from about 15-60%, preferably about 20-55%.

The copolymers which find use in the invention will generally range from about 15-80% of amino acids provided by fibroin units, where the average number of elastin to silk units will range from about 0 to 8.

The polymers will be at least about 15 kDa and generally not more than about 150 kDa, usually not more than about 125 kDa, preferably ranging from about 35-100 kDa. In order to achieve the copolymers, the number of segments will provide for the desired molecular weight. Therefore, the number of segments can vary widely, depending upon the size of each individual segment. Thus, the number of segments may vary from about 2-40, more usually ranging from about 6-20. Based on the method of preparation, there may be non-repetitive units at the N- and C- termini. Usually, the terminal sequences will contribute fewer than ten number percent of the amino acids, more usually fewer than five number percent of the amino acids. Generally, the sequence will range from about 0-125 amino acids, more usually from about 0-60 amino acids, where the total number of amino acids will generally not exceed about 100 amino acids, more usually not exceed about 50 amino acids.

For special applications the polymers may be modified by introducing intervening sequences between segments or blocks of segments, where the total number of repeating units per block may vary from about 4 to 40, thus involving two or more segments. The intervening sequences may include from about 1 to 60, usually about 3 to 40 amino acids, and may provide for a wide variety of properties. For example, by including amino acids which have chemically reactive sidechains, one may provide for sites for linking a variety

of chemically or physiologically active compounds, for cross-linking, for covalently bonding compound which may change the rate of resorption, tensile properties or the like. Thus amino acids, such as cysteine, aspartic acid, glutamic acid, lysine and arginine may be incorporated in these intervening sequences. Alternatively, the sequence may provide for sequences which have physiological activity, such as cell binding, specific protein binding, enzyme substrates, specific receptor binding, and the like. In this manner, the useful properties of the basic protein may be greatly;y varied in accordance with the intended use, being tailored for specific applications.

The polymers have good mechanical properties to form a wide variety of products. The protein polymers may be drawn, molded, cast, spun, extruded, or the like, in accordance with known ways for forming structures such as films, formed objects, fibers, or unformed structures, such as amorphous masses, and the like. Also, gels may be formed which may be shaped in a variety of ways, depending upon the particular application. The compositions can be sterilized by conventional ways to provide sterile products.

The subject compositions can be used to provide a wide variety of devices, such as membranes, sutures, staples, bone pins, screws, wound dressings, and as drug depots, where the products may be formed prior to implantation or in situ . The compositions as formed are found to provide the necessary mechanical properties for the particular applications, the resorption times can be controlled so as to ensure mechanical maintenance during the time required for structure integrity, and at the same time ensuring that the device or material need not be manually removed, since the material undergoes resorption.

The subject compositions may be used in combination with other materials, such as collagen, fibrinogen, and other natural proteins; hyaluronic acid, dextran, or other polysaccharides; or polyethylene oxide, polyhydroxyalkanoates, or other polyesters, to produce

blended materials to provide a larger range of physical and biological properties, for applications, such as wound dressings or membranes for the prevention of surgical adhesions. For example, the protein polymer SELP3 combined with sodium hyaluronate, in equal proportions by weight, may be used to prepare a film, which compared to pure hyaluronate gels, exhibits greater mechanical toughness and a decreased resorption rate.

The compositions may be prepared in accordance with the manner described in U.S. Patent No. 5,243,038. This procedure involves synthesizing small segments of single stranded DNA of from about 15-150 nucleotides to provide a plurality of fragments which have cohesive ends, which may be ligated together to form a segment or a plurality of segments. The first dsDNA fragment is cloned to ensure the appropriate sequence, followed by the addition of successive fragments, which are in turn cloned and characterized, to ensure that the integrity of the sequence is retained. The fragments are joined together to form a "monomer" which then becomes the major repeating building block of the polymer gene.

Alternatively, long single strands may be prepared, cloned and characterized, generally being of at least 100 nucleotides and up to about 300 nucleotides, where the two single strands are hybridized, cloned and characterized and may then serve as the monomer or the building block. The monomers may then be multimerized, having complementary termini, particularly cohesive ends, so that the polymers will have two or more monomers present. The multimers may then be cloned in an appropriate vector and characterized to determine the number of monomers and the desired size polymer selected. Expression can be achieved in an expression host using transcriptional regulatory regions functional in the expression host. The expression host can be prokaryotic or eukaryotic, particularly bacterial, e.g. E. coli , B . subtilis, etc.; yeast, e.g. Saccharomyces, Neurospora, etc.; insect cells, plant cells, and the like.

If desired, a signal sequence may be provided for secretion of the polymer. A wide variety of signal sequences are known and have been used extensively for secreting proteins which are not normally secreted by the expression host. After completion of expression, where the protein is retained in the host, the cells are disrupted and the product extracted from the lysate. Where the product is secreted, the product may be isolated from the supernatant. In either case, various techniques for purifying the products may be employed, depending upon whether the products are soluble or insoluble in the medium. Where insoluble, impurities may be extracted from the polymer, leaving the polymer intact. Where soluble, the polymer may be purified in accordance with conventional ways, such as extraction, chromatography , or the like.

The following examples are offered by way of illustration and not by limitation.

EXPERIMENTAL Example 1. Preparation of polymers.

E. coli strain EC3 containing the respective plasmid encoding each polymer shown in Table 1 below, was prepared in accordance with the methods described in U.S. Patent No. 5,243,038. Each strain was then fermented using a fed- batch method.

Biomass for each polymer was harvested from the fermentation broth by centrifugation in a Sorval RC3B using a H6000A rotor at 5,000 rpm for 30 minutes at 10°C to yield a packed cell paste. 500 grams of cell paste was resuspended in 2 liters of 50 mM Tris buffer (pH=8.0). The cell slurry was homogenized using a Manton Gaulin cell disrupter at 7-8,000 psi with three complete passes of the liquid. The cell homogenate was passed through a chilled heat exchanger to maintain the temperature at 15°C or less. Pancreatic DNAse was added to the homogenate to a final concentration of 1 μg/ml and stirred at room temperature for 2 hours. The homogenate was centrifuged in a Sorval RC3B

centrifuge using a H6000A rotor at 5,000 rpm for 1 hour at 10°C.

For SELPO, 3, 7, and 8, the supernatant was placed into 12-14,000 molecular weight cut-off dialysis bags and dialyzed against 2 changes of lOOx volume of 20 mM sodium acetate buffer (pH=4.7) for 24 hours. The contents of the bags were transferred to centrifuge bottles and centrifuged in a Sorval RC3B centrifuge using a H6000A rotor at 5,000 rpm for 1 hour at 10°C. The supernatant was removed to a large beaker and the pH adjusted to 8.0 by addition of 30% ammonium hydroxide. Saturated ammonium sulfate was then added to reach a final concentration of 20% for SELPO, 25% for SELP8 and 3, and 33% for SELP7. The solution was stirred at room temperature for 1 hour. The solution was centrifuged in a Sorval RC3B using a H6000A rotor at 5,000 rpm for 30 minutes at 10°C. The pellet was resuspended in 2 liters of deionized water, placed in dialysis bags, and dialyzed against 3 changes of deionized water of lOOx volume over 48 hours. The contents of the bags were shell frozen and lyophilized to dryness.

For SELP4 and 5, the centrifuged homogenate supernatant was directly precipitated with ammonium sulfate at a concentration of 25%. The solution was then centrifuged in a Sorval RC3B using a H6000A rotor at 5,000 rpm for 1 hour at 10°C. The pellet was resuspended in 5 liters of 4M LiBr and stirred at 4°C for 16 hours. The solution was centrifuged in a Sorval RC3B centrifuge using a H6000A rotor at 5,000 rpm at 10°C for 1 hour. The pH of the supernatant was adjusted to pH 3.7 by slow addition of 1M acetic acid at 4°C. The solution was centrifuged in a Sorval RC3B using a H6000A rotor at 5,000 rpm at 10°C for 1 hour. The supernatant pH was adjusted to 8.0 by addition of ammonium hydroxide and then dialyzed against 3 changes of lOOx volume deionized water over 48 hours. The solution was removed from dialysis and centrifuged in a Sorval RC3B using a H6000A rotor at 5,000 rpm at 10°C for 1 hour. Saturated ammonium sulfate was added to the supernatant to reach 25%

of saturation and stirred for 1 hour. The solution was centrifuged in a Sorval RC3B using a H6000A rotor at 5,000 rpm at 10°C for 1 hour. The pellet was dissolved in 4.5M LiBr, placed in dialysis bags, and dialyzed against 3 changes of lOOx volume of deionized water. The contents of the bags were shell frozen and lyophilized to dryness.

All reagent solutions used in the following procedures were depyrogenated prior to use by filtration through a 10,000 nominal molecular weight cut-off hollow fiber cartridge (AG Technologies) . All glassware and utensils used were sterilized and depyrogenated by heating at 180°C for 4 hours. 4-5 grams of all SELP dried polymers were dissolved in 1.2 liters of 10M urea. 20 mis of 2M Tris pH8.0 and 780 mis of milli-Q water were added. The solution was sonicated to promote full dissolution of the protein. 500 grams of Whatman DE52 ion exchange resin was prepared by precycling through acid and base treatment as recommended by manufacturer prior to and in between each usage. The resin was finally equilibrated with 6M urea, 20 mM Tris pH8.0 in a beaker with gentle stirring. The resin was filtered in a buchner funnel until excessive liquid was removed. The cake of resin was placed in a beaker and the protein solution was added. The slurry was stirred gently for 1 hour. The slurry was filtered in a buchner funnel and the liquid was collected in a cleaned vacuum flask. 500 grams of fresh precycled and equilibrated resin was added to a clean beaker and the filtered solution was added.. The slurry was stirred gently for 1 hour and filtered again. The filtered solution was once more combined with 500 grams of freshly precycled and equilibrated resin, stirred for 1 hour, and filtered. The final filtered solution was placed in 6,000 molecular weight cut-off dialysis bags which had been soaked in 0.5N NaOH for at least 24 hours. The solution was dialyzed against 3 changes of lOOx volume of deionized water. The dialyzed solution was removed from the bags, placed in depyrogenated lyophilization flasks and lyophilized to

dryness. Employing the above procedure, the following polymers were prepared.

TABLE 1

Polymer (MW) Polymer Block Sequence ¹ Domain Abbr. ² E/S ³ %S*

SELPO (80,502) [(VPGVG GAGAGS , E8S2 4.0 21.9 (SEQ ID NO:03)

SELP8 (69,934) [(VPGVG) ₈ (GAGAGS) ₄ ] ₁₃ E8S4 2.0 35.3 SEQ ID NO:04)

SELP7 (80,338) [(VPGVG) ₈ (GAGAGS)J _I3 E8S6 1.33 45.0 (SEQ ID NO:05)

SELP3 (84,267) [(VPGVG) ₈ (GAGAGS) ₈ ] ₁₂ E8S8 1.0 51.9 (SEQ ID NO:06)

SELP4 (79,574) [(VPGVG) ₁₂ (GAGAGS) ₈ ], E12S8 1.5 42.2 (SEQ ID NO.07)

SELP5 (84,557) [(VPGVG) ₁₆ (GAGAGS) ₈ ] ₈ E16S8 2.0 35.7 (SEQ ID NO:08)

The first and last block domain of each polymer is split within the silk blocks such that both parts sum to a whole domain. All polymers also contain an additional head and tail sequence which constitutes approximately 6% of the total amino acids.

Designates the number of consecutive blocks per repeating domain

(E = elastin-like block, S = silk-like block)

Ratio of blocks per polymer.

% of total amino acids in polymer contributed by silk-like blocks.

Other polymers which were prepared include [ (VPGVG) ₃₂ (GAGAGS) ₈ ] (SEQ ID NO:09), referred to as SELP6. Example 2. SELP films.

SELP films that were approximately 0.05 mm thickness were produced by solvent evaporation.

Approximately 1.7 grams of each polymer, except for SELP7 where only 1.05 grams was used, were solubilized in 34 mis of 88% formic acid. The solution was stirred for 7 hours at room temperature to insure complete solubilization. The solution was then poured into a film casting apparatus consisting essentially of a rectangular polyethylene trough with a removable polyethylene bottom. The casting apparatus was placed in a vacuum oven attached to a nitrogen gas source for sparging the atmosphere. The films were dried in

the sealed oven drawing a 10-15 micron vacuum with a slow continual influx of nitrogen gas at 60-75° C. After 15-18 hours of drying, the apparatus was disassembled and the film was peeled off the polyethylene bottom. The films were exposed for 5 minutes to a basic atmosphere (5% open solution of ammonium hydroxide in a sealed desiccator) to neutralize any residual formic acid.

A polyethylene sheet of the same area dimensions as the protein film was roughened by hand using fine grit sand paper and a fine film of cyanoacrylate glue was spread over its surface. The protein film was applied to the wet surface. A teflon sheet was placed on top and bottom of the polyethylene and protein layers and stainless steel plates were placed around those. The entire assembly was pressed in a Carver laboratory press at a force of 0.8 metric tons for 18 hours at room temperature. The polyethylene/protein film laminated sheet was placed on a cutting board and 1.3 cm diameter discs were punched out using a stainless steel punch and rubber mallet. The discs were placed individually in stoppered glass vials.

Specimens were produced from each of the polymers as well as denatured collagen protein (DCP) produced identically as described for the SELP films. Bovine collagen (fibrillar form, lot number 921101) was obtained from Colla-Tec, Inc. (Plainsboro, New Jersey) . It was completely solubilized in 88% formic acid producing a clear but viscous solution. All specimens were sterilized by electron beam irradiation at 2.5 +/- 0.2 Mrads. Each disk was implanted subcutaneously in the back of rats such that the protein film was in direct contact with the muscle tissue. The specimens remained in the animals for different periods of time: one, four and seven weeks post implantation. At each time interval six specimens per polymer group were retrieved for protein analysis. Additional specimens from each group were evaluated for tissue reaction by histology.

Non-implanted and retrieved specimens were analyzed to determine the mass of SELP film contained per specimen. Amino acid analysis was performed on each specimen by sealing them individually in an hydrolysis vial with constant boiling hydrochloric acid and heating for 24 hr at 100-110°C. After hydrolysis, the specimen was extracted and an aliquot of the extract was derivatized with PTC. The derivatized amino acids were separated by reverse phase HPLC and quantified by their absorbance at 254 nm according to the methods of Henrickson and Meredith (Anal. Biochem . 137, 65-74, 1984).

The mass of the SELP film present on each specimen was determined. The amino acid contribution of the SELP protein was estimated based on the total content of the amino acids G,A,S,V and P which for the pure polymers is >95%. Other amino acids potentially contributed by extraneous protein deposited onto the specimens during residence in the body were excluded from these analyses. Average SELP film mass for non-implanted specimens was determined from the same batch of specimens used for implantation. Average SELP film mass for retrieved specimens was similarly calculated except that replicates having values greater than two standard deviations from the mean were discarded. Deviations in many cases were due to partial retrieval of specimens that had fragmented in the tissue after implantation and may not reflect true resorption.

Resorption Analysis and Results

Resorption analysis was conducted statistically by analyzing four specimen population treatment groups. These were: (1) non-implanted; (2) one week post-implantation;

(3) four weeks post-implantation; and (4) seven weeks post-implantation.

TABLE 2 Polymer Film Mass Remaining as Determined by AA Composition Analysis (in milligrams)

SELPO SELP3 SELP4 SELP5

Initial Film 12.21 +/-1.41 5.99 +/-0.46 8.19 +/-0.86 8.51 +/-1.04 Mass

1 Week Film 0.53 +/-0.31 5.93 +/-0.73 7.89 +/-0.55 7.72 +/-1.57 Mass

4 Week Film 0.27 +/-0.13 6.24 +/-0.61 9.20 +/-1.08 7.49 +/-0.75 Mass

7 Week Film 0.10 +/-0.02 3.49 +/-1.60 8.56 +/-0.67 8.77 +/-0.97 Mass

SELP7 SELP8 DCP

Initial Film 3.27 +/-0.34 8.43 +/-0.59 6.6 +/-1.04 Mass

1 Week Film 4.67 +/-1.33 1 1.13 +/- 1.40 0.15 +/-0.07 Mass

4 Week Film 0.19 +/-0.16 8.26 +/-1.21 0.09 +/-0.03 Mass

7 Week Film 0.08 +/-0.03 1.52 +/-1.40 0.07 +/-0.03 Mass

TABLE 3 Polymer Film Remaining as Percent of Non-implanted Mass

SELPO SELP3 SELP4 SELP5 SELP7 SELP8 DCP

Initial ilm 100.0% 100.0% 100.0% 100.0% 100.0% 100.0% 100.0% Mass

1 Week 4.3% 98.9% 96.3 % 90.7% 142.8% 132.0% 2.3%

Film

Mass

4 Week 2.2% 104.1 % 1 12.4% 88.0% 5.8% 98.0% 1.3%

Film

Mass

7 Week 0.8% 58.2% 104.5 % 103.1 % 2.6% 18.1 % 1.1 %

Film

Mass

The results from Table 2 are the values for the mass of protein film contained on specimens after implantation. Each value is the mean of at least five specimen masses as determined by amino acid composition. Table 3 displays the same values as a percent of the initial weight prior to

implantation as determined by the mean mass of six specimens of the non-implanted specimens. The results indicate that upon implantation, SELPO and DCP are substantially resorbed by one week, falling below 5% of their non-implanted masses. SELP7 is substantially resorbed by four weeks with only 5.8% remaining. SELP8 and SELP3 are resorbing by seven weeks with mean values of 18.1% and 58.2% remaining, respectively. SELP4 and SELP5 films show no evidence of resorption at seven weeks. From the above results one may conclude the following. Faster resorption correlates with compositions containing domains of silk-like blocks fewer than eight. The polymers containing eight silk-like blocks have substantially reduced rates of resorption. However, the total content of silk-like blocks in the copolymer composition does not correlate with resorption rate. While very similar compositionally, SELP7 and SELP8 resorbed quickly, while SELP4 and SELP5 do not resorb in seven weeks. The lack of resorption of SELP4 and SELP5 films at seven weeks post-implantation corresponds with repeating domains containing greater than eight elastin-like blocks. Although their silk-like block lengths are identical at eight, SELP4 and 5 with elastin-like block lengths of 12 and 16 resorb to a lesser degree than SELP3, which has an elastin-like block length of 8.

The subject polymers, regardless of their composition, form free-standing films with strength enough to allow easy handling. SELP7 and SELP4 films have tensile strengths of 19+/-1 and 21+/-8 MPa, respectively. The compositional difference between them that causes SELP7 to resorb in four weeks and SELP4 to remain intact beyond seven weeks makes little apparent difference in their tensile properties. These strengths are adequate for their use in surgical and wound healing applications. The observed resorption of these polymers occurs via surface erosion. This is consistent with the mechanism of degradation of SELP proteins within the body. At

physiological conditions, proteins will degrade only through the action of proteases. Because endogenous proteases are high molecular weight compounds of approximately 20 kDa or greater, their diffusion into the dense SELP films will be limited. The degradation of SELP films is, therefore, progressive from the external surfaces of the material. The subject materials therefore should undergo a slow loss of mechanical integrity while being reduced in mass.

Example 2: SELP Porous Sponges

The Function of an implanted material depends greatly on its form, morphology, and mechanical strength, SELP polymers have been fashioned into a variety of forms; dense films, porous sponges, and fibrillar mats. Dense films or sheets, as described above, are semi-permeable barriers which may have utility in surgical repairs by restricting fluid or gas flow, blocking cellular migration, maintaining tissue separations, and confining and protecting implanted organs or devices. Their properties will vary depending on their permeability and their thickness which may range from 0.05 mm to greater than 1 mm. For example varying their thickness will effect their mechanical strength, their resistance to abrasion, and their ultimate resorption.

SELP polymers have been produced as three dimensional, porous sponges to serve as implantable materials that will support cell and tissue ingrowth. Preparation of SELP5 Sponges.

All glassware to come in contact with the protein polymer was depyrogenated by heating to 180°C for 6 hours. SELP5 (0.978 g) was stirred in LAL reagent grade water until dissolved to yield a 1.0% w/v aqueous solution. This solution was aseptically transferred to a 100ml ST 24/40 pear shaped flask and tared. This flask was fitted with a spray trap, attached to a rotary evaporator, and 65.2 g of water was evaporated using a bath temperature of 39°C, a system pressure of 42 mbar, and a rotation rate of 125 rpm, to yield a solution of 3.0% w/v concentration. This solution

was poured 6mm deep into six standard sterilized Petri dishes (mm diameter) ; covered with standard lids; placed on a small plastic tray; and placed in a - 8°C freezer overnight. After freezing, the lids were removed from the Petri dishes; the Petri dishes were placed into a 1200 ml wide mouth lyophilization flask and lyophilized to dryness. After completion of lyophilization, the sponges were removed from their Petri dishes and placed, individually, into a 100ml wide mouth flask containing 75ml of methanol at room temperature. The head space was evacuated to less than the vapor pressure of the methanol to induce eubulation and insure compete displacement of air entrained within the sponge by the methanol. The sponge, wetted with methanol was allowed to stand for 5 minutes at room temperature at room temperature, methanol was removed from the sponge by washing 6 times with LAL reagent grade water (175ml per wash) and allowing each was to stand for 5 minutes. The sponges, wetted by water, were returned to 35mm diameter Petri dishes, frozen at -8°C, and again lyophilized. The lyophilized sponges were placed into new 35mm diameter Petri dishes, lids applied and sealed with parafilm ^® , placed into a plastic instrument bag, heat sealed, and sterilized using an electron beam irradiation at 2.8 Mrads.

The sponges were dimensionally stable when immersed in saline or water. When engorged with saline, the sponge turned from white to grey and was somewhat translucent. The engorged sponge retained its original dimensions. Minimal swelling was observed. The geometry and edges of the wet sponge remained unchanged. The observed aqueous stability of the SELP 5 sponges is different from the properties of collagen hemostatic sponges (Helistat, Marion Laboratories, Kansas City, MO) which almost immediately collapse when exposed to liquid.

SELP5 sponges were cut into 2 x 2 x 0.4 cm specimens and applied to 2 x2 cm full thickness dermal wounds in pigs. 2 x 2 x 0.3 cm specimens of Helistat were similarly applied to wounds. After bleeding was controlled and the wound flushed

with saline, the specimens were laid into the tissue void such that they would firmly contact the wound bed. The Helistat specimens became completely or partially engorged within a few seconds to several minutes after application depending on the amount of the blood in the wound. The engorged Helistat specimens collapsed and shrunk resulting in nonuniform coverage of the wound, in some cases, exposing part of the wound beds.

The SELP5 sponges remained substantially white during the 5 minute observation period after application indicating that they did not immediately absorb blood. One corner of one specimen turned red within a minute after application. It remained physically unchanged. The SELP5 sponges adhered well to the wound bed and could not be lifted out of the wound with forceps using mild tension. The SELP5 sponges did not shrink upon contact with the bloody tissue and continued to completely cover the wound during observation.

All wounds were covered with petrolatum gauze pads and bandaged. After 7 days, the wounds were undressed and observed to determine the extent of healing. Wounds containing SELP5 sponges had progressed normally through the healing process as compared to wounds to which no material was applied. The sponge material had not been extruded from the wound as there was no evidence of extraneous material on the gauze pads. No evidence of excessive inflammation was observed. Epithelialization of the wound was in progress.

Example 3: SELP Fibrous Meshes

SELP polymers can be fabricated as non-woven fibrous meshes to produce fibrillar mats which are flexible, have good drapability, and are stable in wet environments. Fibrous meshes with similar physical properties were produced from SELP5, SELP7, and SELPF using the following procedure. 1 gram of polymer was dissolved in 88% formic acid with stirring at room temperature until homogenous. For SELP5, 5 mis of formic acid were used to dissolve the

lyophilized polymer. For SELP7 and SELPF, 4 and 3 mis of formic acid were used, respectively.

The polymer dope was drawn into a lcc polypropylene syringe, affixed with a 75mm x 20 gauge stainless steel hypodermic needle, and mounted on a Sage Instruments syringe pump (model 341B) . The pump was set to deliver approximately 0.05 to 0.07 cc/minute. The tip of the needle was placed at 90° to a gas stream delivered from a stainless steel needle (25mm x 20 gauge) . A more acute angle was also used. The dope delivery needle and the gas delivery needle were mounted onto a steel "L"-bracket using miniature "C"-clamps and pads of neoprene rubber such that a gap of 1 mm separated their tips. The tips were displaced in the vertical direction by 0.5 mm such that the gas stream passed slightly over the flanged end of the hypodermic needle. The gas stream was supplied either with compressed air or high purity (extra dry) nitrogen gas. Compressed air was supplied by an oiless compressor using a diaphragm pump. The air in the reservoir was a ca. 8 atm pressure and was regulated down to ca. 2-6 atm before being fed to the spray apparatus. When nitrogen was used, it was delivered at 20 psi. The relative humidity was less than 47%.

Fine filaments were formed on and around the edges of a rectangular, 1/16 inch polypropylene mesh that was used as a target approximately 7-12 inches from needle tips. Filaments streamed off the edges of the target and when they were approximately 5 cm in length, they were collected on a circular, metal wire loop of 38 mm in diameter. Filaments were collected across the loop forming a web of suspended filaments in the center. The web was removed from the loop by compressing the web between two 35mm polystyrene discs and pressing the web through the wire frame. Fibrous meshes were built up by compressing 5-8 webs between the same discs. The meshes were stabilized by flooding them with 1 ml of either 100% methanol or 100% ethanol and allowing them to dry under ambient conditions. The meshes were sterilized by

electron beam irradiation at a dose of 2.5 MRads. Under microscopic observation, the meshes consisted of fine filaments which varied in diameter from 0.1 to lOμm. The meshes were stable when placed in saline for more than 24 hours.

The meshes were applied to 2 x 2 cm partial and full thickness dermal wounds in pigs in order to investigate their biocompatibility and their ability to incorporate within the healing tissue. The meshes were removed from the polystyrene discs with forceps and applied to the wound bed. The edges of the meshes could be pulled across the tissue allowing the mesh to be spread and/or rearranged over the wound. The wounds were covered and examined every two days for signs of bioincompatibility. No adverse effects were observed in wounds containing SELP fibrous meshes. After 14 days, the wounds were completely epithelialized. Histological examination of tissues from wounds to which SELPF fibrous webs had been applied showed that foreign material in the form of filaments had been incorporated into the healing tissue.

These data indicate that SELP fibrous meshes are well tolerated in healing tissue. Their presence does not interfere with normal healing. In one case, SELP filaments were clearly shown to reside within the healed tissue. SELP films, meshes, and sponges can serve as resorbable packing materials that can be used to augment the loss of soft tissue that occurs during traumatic injury or surgical disection. Their application at the time of injury can encourage infiltration, overgrowth, and eventual replacement of the materials with healthy tissue. The mass of the implanted material can provide enough stability to maintain the geometric contours of the body site at which the tissue was lost. Their presence can also mechanically reinforce the wound site such that delicate, healing tissues can form while protected from further physical injury.

It is evident from the above results, that the subject compositions have particularly desirable properties for uses

in plants. By varying compositional ratios, the rate of resorption can be varied greatly, without significant changes in tensile properties. The compositions can be formed in a wide variety of devices or objects, to find extensive use for a variety of purposes and context as implants.

All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.

Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.

SEQUENCE LISTING

(1) GENERAL INFORMATION:

(i) APPLICANT: Protein Polymer Technologies, Inc. (ii) TITLE OF INVENTION: Synthetic Proteins As Implantables (iii) NUMBER OF SEQUENCES: 9

(iv) CORRESPONDENCE ADDRESS:

(A) ADDRESSEE: Flehr, Hohbach, Test, Albritton & Herbert

(B) STREET: Four Embarcadero Center, Suite 3400

(C) CITY: San Francisco

(D) STATE: CA

(E) COUNTRY: U.S.A.

(F) ZIP: 94111-4187

(v) COMPUTER READABLE FORM:

(A) MEDIUM TYPE: Floppy disk

(B) COMPUTER: IBM PC compatible

(C) OPERATING SYSTEM: PC-DOS/MS-DOS

(D) SOFTWARE: PatentIn Release #1.0, Version #1.25

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER: PCT/US95/

(B) FILING DATE:

(C) CLASSIFICATION:

(Viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: Rowland, Bertram I

(B) REGISTRATION NUMBER: 20,015

(C) REFERENCE/DOCKET NUMBER: FP-58847-l-PC/BIR

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: 415-781-1989

(B) TELEFAX: 415-398-3249

(2) INFORMATION FOR SEQ ID NO:l:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 6 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l:

Gly Ala Gly Ala Gly Ser 1 5

(2) INFORMATION FOR SEQ ID NO:2:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 5 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:

Val Pro Gly Val Gly 1 5

(2) INFORMATION FOR SEQ ID NO:3:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 936 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 35 40 45

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 50 55 60

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 65 70 75 80

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 85 90 95

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 100 105 110

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 115 120 125

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 130 135 140

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 145 150 155 160

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 165 170 175

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 180 185 190

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 195 200 205

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 210 215 220

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 225 230 235 240

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 245 250 255

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 260 265 270

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 275 280 285

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 290 295 300

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 305 310 315 320

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 325 330 335

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 340 345 350

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 355 360 365

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 370 375 380

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 385 390 395 400

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 405 410 415

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 420 425 430

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 435 440 445

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 450 455 460

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 465 470 475 480

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 485 490 495

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 500 505 510

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 515 520 525

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 530 535 540

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 545 550 555 560

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 565 570 575

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 580 585 590

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 595 600 605

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 610 615 620

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val

625 630 635 640

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 645 650 655

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 660 665 670

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 675 680 685

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 690 695 700

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 705 710 715 720

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 725 730 735

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 740 745 750

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 755 760 765

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 770 775 780

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 785 790 795 800

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 805 810 815

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 820 825 830

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 835 840 845

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 850 855 860

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 865 870 875 880

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 885 890 895

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 900 905 910

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 915 920 925

Gly Ser Gly Ala Gly Ala Gly Ser 930 935

(2) INFORMATION FOR SEQ ID NO:4:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 832 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 35 40 45

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 50 55 60

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 65 70 75 80

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 85 90 95

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 100 105 110

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 115 120 125

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 130 135 140

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 145 150 155 160

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 165 170 175

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 180 185 190

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 195 200 205

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 210 215 220

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 225 230 235 240

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 245 250 255

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 260 265 270

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 2 27755 228800 228855

Gly Gly Ser Gly Ala

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 305 310 315 320

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val

325 330 335

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 340 345 350

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 355 360 365

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 370 375 380

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 385 390 395 400

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 405 410 415

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 420 425 430

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 435 440 445

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 450 455 460

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 465 470 475 480

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 485 490 495

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 500 505 510

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 515 520 525

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 530 535 540

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 545 550 555 560

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 565 570 575

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 580 585 590

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 595 600 605

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 610 615 620

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 625 630 635 640

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 645 650 655

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 660 665 670

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 675 680 685

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 690 695 700

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 705 710 715 720

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 725 730 735

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 740 745 750

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 755 760 765

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 770 775 780

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 785 790 795 800

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 805 810 815

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 820 825 830

(2) INFORMATION FOR SEQ ID NO:5:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 988 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 35 40 45

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 50 55 60

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 65 70 75 80

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 85 90 95

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 100 105 110

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 115 120 125

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala

130 135 140

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 145 150 155 160

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 165 170 175

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 180 185 190

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 195 200 205

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 210 215 220

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 225 230 235 240

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 245 250 255

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 260 265 270

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 275 280 285

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 290 295 300

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 305 310 315 320

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 325 330 335

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 340 345 350

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 355 360 365

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 370 375 380

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 385 390 395 400

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 405 410 415

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 420 425 430

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 435 440 445

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 450 455 460

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 465 470 475 480

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 485 490 495

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 500 505 510

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 515 520 525

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 530 535 540

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 545 550 555 560

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 565 570 575

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 580 585 590

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 595 600 605

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 610 615 620

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 625 630 635 640

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 645 650 655

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 660 665 670

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 675 680 685

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 690 695 700

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 705 710 715 720

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 725 730 735

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 740 745 750

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 755 760 765

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 770 775 780

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 785 790 795 800

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 805 810 815

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 820 825 830

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 835 840 845

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly

850 855 860

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 865 870 875 880

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 885 890 895

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 900 905 910

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 915 920 925

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 930 935 940

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 945 950 955 960

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 965 970 975

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 980 985

(2) INFORMATION FOR SEQ ID NO:6:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1056 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 35 40 45

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 50 55 60

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 65 70 75 80

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 85 90 95

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 100 105 110

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 115 120 125

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 130 135 140

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 145 150 155 160

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 165 170 175

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 180 185 190

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 195 200 205

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 210 215 220

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 225 230 235 240

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 245 250 255

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 260 265 270

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 275 280 285

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 290 295 300

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 305 310 315 320

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 325 330 335

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 340 345 350

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 355 360 365

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 370 375 380

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 385 390 395 400

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 405 410 415

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 420 425 430

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 435 440 445

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 450 455 460

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 465 470 475 480

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 485 490 495

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala

500 505 510

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 515 520 525

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 530 535 540

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 545 550 555 560

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 565 570 575

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 580 585 590

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 595 600 605

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 610 615 620

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 625 630 635 640

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 645 650 655

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 660 665 670

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 675 680 685

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 690 695 700

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 705 710 715 720

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 725 730 735

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 740 745 750

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 755 760 765

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 770 775 780

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 785 790 795 800

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 805 810 815

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 820 825 830

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 835 840 845

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 850 855 860

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 865 870 875 880

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 885 890 895

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 900 905 910

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 915 920 925

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 930 935 940

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 945 950 955 960

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 965 970 975

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 980 985 990

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 995 1000 1005

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 1010 1015 1020

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 1025 1030 1035 1040

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser iriAc men inee

(2) INFORMATION FOR SEQ ID NO:7:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 972 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 35 40 45

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 50 55 60

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 65 70 75 80

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser

85 90 95

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 100 105 110

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 115 120 125

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 130 135 140

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 145 150 155 160

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 165 170 175

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 180 185 190

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 195 200 205

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 210 215 220

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 225 230 235 240

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 245 250 255

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 260 265 270

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 275 280 285

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 290 295 300

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 305 310 315 320

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 325 330 335

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 340 345 350

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 355 360 365

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 370 375 380

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 385 390 395 400

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 405 410 415

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 420 425 430

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 435 440 445

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 450 455 460

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 465 470 475 480

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 485 490 495

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 500 505 510

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 515 520 525

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val 530 535 540

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 545 550 555 560

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 565 570 575

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 580 585 590

Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala 595 600 605

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 610 615 620

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 625 630 635 640

Gly Ser Gly Ala Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly 645 650 655

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 660 665 670

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 675 680 685

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 690 695 700

Pro Gly Val Gly Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 705 710 715 720

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 725 730 735

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 740 745 750

Gly Ala Gly Ser Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 755 760 765

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 770 775 780

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 785 790 795 800

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly

805 810 815

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 820 825 830

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 835 840 845

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 850 855 860

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 865 870 875 880

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 885 890 895

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 900 905 910

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Gly Ala Gly Ala 915 920 925

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 930 935 940

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 945 950 955 960

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 965 970

(2) INFORMATION FOR SEQ ID NO:8:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1024 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: peptide

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 • 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 35 40 45

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 50 55 60

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 65 70 75 80

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 85 90 95

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 100 105 110

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 115 120 125

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 130 135 140

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 145 150 155 160

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 165 170 175

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 180 185 190

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 195 200 205

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 210 215 220

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 225 230 235 240

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 245 250 255

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 260 265 270

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 275 280 285

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 290 295 300

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 305 310 315 320

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 325 330 335

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 340 345 350

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 355 360 365

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 370 375 380

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 385 390 395 400

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 405 410 415

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 420 425 430

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 435 440 445

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 450 455 460

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala

465 470 475 480

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 485 490 495

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 500 505 510

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 515 520 525

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 530 535 540

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 545 550 555 560

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 565 570 575

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 580 585 590

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 595 600 605

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 610 615 620

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 625 630 635 640

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 645 650 655

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 660 665 670

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 675 680 685

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 690 695 700

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 705 710 715 720

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 725 730 735

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 740 745 750

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 755 760 765

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 770 775 780

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 785 790 795 800

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 805 810 815

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 820 825 830

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 835 840 845

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 850 855 860

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 865 870 875 880

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 885 890 895

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 900 905 910

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 915 920 925

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 930 935 940

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 945 950 955 960

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 965 970 975

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 980 985 990

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 995 1000 1005

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 1010 1015 1020

(2) INFORMATION FOR SEQ ID NO:9:

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 208 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val 1 5 10 15

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 20 25 30

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 35 40 45

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 50 55 60

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 65 70 75 80

Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val

85 90 95

Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro 100 105 110

Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly 115 120 125

Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val 130 135 140

Gly Val Pro Gly Val Gly Val Pro Gly Val Gly Val Pro Gly Val Gly 145 150 155 160

Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala 165 170 175

Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala 180 185 190

Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser Gly Ala Gly Ala Gly Ser 195 200 205