Claims
- 1. A substantially purified polypeptide selected from the group consisting of:
(a) a polypeptide comprising an amino acid sequence of SEQ ID NO:1, 3, 4, 6, 8, or 10; (b) soluble fragments of the polypeptide of (a) having disintegrin activity; (c) fragments of the polypeptide of (a) comprising at least 20 to 30 contiguous amino acids; (d) fragments of the polypeptide of (a) having a disintegrin activity; (e) fragments of the polypeptide of (a) comprising a disintegrin domain amino acid sequence; (f) SEQ ID NO:6 from about amino acid 73 to about an amino acid between about 360 and 362; (g) SEQ ID NO:8 from an amino acid between about residue 1 and 16 to about an amino acid between 285 and 287; (h) SEQ ID NO:10 from an amino acid between about residue 1 and 73 to an amino acid between about residue 314 and 329; (i) amino acid sequences comprising at least 10 continguous amino acids and sharing amino acid identity with the amino acid sequences of (a)-(h), wherein the percent amino acid identity is selected from the group consisting of at least 85%, at least 90%, at least 95%, at least 97.5%, at least 99%, and at least 99.5%; and (j) a polypeptide comprising an amino acid sequence of SEQ ID NO :25.
- 2. A soluble polypeptide having disintegrin activity according to claim 1 linked to a second polypeptide, wherein the second polypeptide is a leucine zipper polypeptide, an Fc polypeptide, or a peptide linker.
- 3. An isolated polynucleotide encoding a polypeptide of claim 1 or 2.
- 4. An isolated polynucleotide selected from the group consisting of:
a) a polynucleotide comprising a sequence of SEQ ID NO:2, 5, 7, or 9; b) a polynucleotide comprising a sequence of SEQ ID NO:5 from about nucleotide 248 to nucleotide 1111 of SEQ ID NO:5; c) a polynucleotide comprising a sequence of SEQ ID NO:7 from a nucleotide between about 82 and 127 to about nucleic acid 936; d) a polynucleotide comprising a sequence of SEQ ID NO:9 from a nucleotide between about 32 and 248 to a nucleotide between about 973 and 1018; e) a polynucleotide that hybridizes under moderately stringent conditions to a polynucleotide comprising the sequence of a), b), c), or d); f) a nucleotide sequence complementary to a sequence of SEQ ID NO:2, 5, 7, or 9; and g) any of nucleotide sequences of a) to f) wherein T can also be U.
- 5. An isolated polynucleotide comprising a sequence of claim 5 operably linked to a polynucleotide encoding a polypeptide of interest.
- 6. An expression vector comprising a polynucleotide of claim 3, 4, or 5.
- 7. A recombinant host cell comprising polynucleotide of claim 3, 4, or 5.
- 8. A method for producing a polypeptide, comprising culturing the host cell of claim 7 under conditions promoting expression of the polypeptide.
- 9. The method of claim 8, further comprising purifying the polypeptide.
- 10. A polypeptide produced by culturing the host cell of claim 7 under conditions to promote expression of the polypeptide.
- 11. A substantially purified antibody that specifically binds to a polypeptide of claim 1.
- 12. The antibody of claim 11, wherein the antibody is a monoclonal antibody.
- 13. The antibody of claim 11, wherein the antibody is a human or humanized antibody.
- 14. The antibody of claim 11, wherein the antibody inhibits the biological activity of the polypeptide of claim 1.
- 15. A method of designing an inhibitor or binding agent of a polypeptide of claim 1, comprising determining the three-dimensional structure of the polypeptide, analyzing the three-dimensional structure for binding sites of substrates or ligands, designing a molecule that is predicted to interact with the polypeptide, and determining the inhibitory or binding activity of the molecule.
- 16. A method for identifying an agent that modulates an activity of a polypeptide of claim 1, comprising:
(a) contacting the agent with a polypeptide of claim 1 under conditions such that the agent and polypeptide interact; and (b) determining the activity of the polypeptide in the presence of the agent compared to a control, wherein a change in activity is indicative of an agent that modulates the polypeptide's activity.
- 17. The method of claim 16, wherein the agent is selected from the group consisting of an antibody, a small molecule, a peptide, and a peptidomimetic.
- 18. A method of inhibiting angiogenesis in a mammal in need of such treatment, comprising administering to the mammal an inhibition-effective amount of a soluble ADAM-H9 disintegrin domain polypeptide.
- 19. The method of claim 18, wherein the soluble ADAM-H9 disintegrin domain polypeptide comprises a sequence selected from the consisting of:
(a) SEQ ID NO:6 from about amino acid 73 to amino acid 360 or 362; (b) SEQ ID NO:8 from an amino acid between about residue 1 and 16 to amino acid 285 or 287; (c) SEQ ID NO: 10 from an amino acid between about residue 1 and 73 to an amino acid between about residue 314 and 329; and (d) fragments of (a)-(c) having disintegrin activity.
- 20. A method for modulating angiogenesis in a tissue, comprising contacting the tissue with a polypeptide of claim 1.
- 21. A method for modulating endothelial cell migration, comprising contacting an endothelial cell with a polypeptide of claim 1.
- 22. The method of claim 20 or 21, wherein the contacting is in vitro.
- 23. The method of claim 20 or 21, wherein the contacting is in vivo.
- 24. A method of inhibiting the binding of an integrin to a ligand comprising contacting a cell that expresses the integrin with an effective amount of a soluble ADAM-H9 disintegrin domain polypeptide.
- 25. The method of claim 24, wherein the soluble ADAM-H9 disintegrin domain polypeptide comprises a sequence selected from the consisting of:
(a) SEQ ID NO:6 from about amino acid 73 to amino acid 360 or 362; (b) SEQ ID NO:8 from an amino acid between about residue 1 and 16 to amino acid 285 or 287; (c) SEQ ID NO:10 from an amino acid between about residue 1 and 73 to an amino acid between about residue 314 and 329; and (d) fragments of (a)-(c) having disintegrin activity.
- 26. A method of modulating the binding of an integrin to a ligand in a mammal in need of such treatment comprising administering an effective amount of a soluble ADAM-H9 disintegrin domain polypeptide.
- 27. The method of claim 26, wherein the mammal is afflicted with a condition selected from the group consisting of ocular disorders; malignant and metastatic conditions; inflammatory diseases; osteoporosis, accelerated bone resorption disorders; restenosis; inappropriate platelet activation, recruitment, or aggregation; thrombosis; and a condition requiring tissue repair or wound healing.
- 28. The method of claim 26, wherein the soluble ADAM-H9 disintegrin domain polypeptide comprises a sequence selected from the group consisting of:
(a) SEQ ID NO:6 from about amino acid 73 to amino acid 360 or 362; (b) SEQ ID NO:8 from an amino acid between about residue 1 and 16 to amino acid 285 or 287; (c) SEQ ID NO: 10 from an amino acid between about residue 1 and 73 to an amino acid between about residue 314 and 329; and (d) Fragments of (a)-(c) having disintegrin activity.
- 29. The method of claim 18, 24, or 26, wherein the soluble ADAM-H9 disintegrin domain is in the form of a multimer.
- 30. The method of claim 29, wherein the multimer is a dimer or trimer.
- 31. The method of claim 30, wherein the multimer comprises an Fc polypeptide, a leucine zipper, or a peptide linker.
- 32. The method of claim 31, wherein the multimer comprises a sequence as set forth in SEQ ID NO:25.
- 33. A system for analyzing polypeptides or polynucleotides comprising:
a data set representing a set of one or more polypeptides of claim 1, or one or more polynucleotides of claim 4, or a combination of polypeptides and polynucleotides; a computer; and a computer algorithm in an executable format on the computer for analyzing the polypeptides or polynucleotides.
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit under 35 U.S.C. § 119 of U.S. Provisional Application Serial No. 60/221,838, filed Jul. 28, 2000, and U.S. Provisional Application Serial No. 60/282,550, filed Apr. 9, 2001, the disclosures of which are incorporated herein by reference.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US01/23709 |
7/27/2001 |
WO |
|