Claims
- 1. An isolated and purified polypeptide from Agkistrodon rhodostoma having an amino acid sequence set forth in SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6, in which Xaa in the amino acid sequences represents a residue of a natural .alpha.-amino acid.
- 2. An isolated and purified polypeptide obtained from the crude venom of Agkistrodon rhodostoma by chromatography on matrex red A-Sepharose.RTM. ion-exchange medium and arginine-Sepharose.RTM. ion-exchange medium and subsequent chromatofocusing with the gel material PBE.RTM. 94, which exhibits a main band with a molecular weight of about 42,000 dalton on SDS-PAGE, has a fibrinogenase activity of about 500 U/mg and has the N-terminal amino acid sequence shown in SEQ ID NO: 7.
- 3. An isolated and purified DNA sequence which codes for a polypeptide as defined in claim 1.
- 4. A recombinant DNA molecule comprising a DNA sequence as defined in claim 3 which is functionally connected to an expression control system which makes expression possible in suitable host systems.
- 5. A recombinant DNA molecule as defined in claim 4, wherein the expression control sequence is an E.coli promoter system, a promoter system of an E.coli bacteriophage, a yeast or fungus expression control sequence or another eukaryotic expression control sequence.
- 6. A host cell which contains at least one recombinant DNA molecule as defined in claim 5.
- 7. A host cell of claim 6, wherein said cell is a bacterial, yeast, fungal, animal or human cell.
- 8. A method for producing a polypeptide having an amino acid sequence set forth in SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5 or SEQ ID NO: 6 which comprises culturing a cell of claim 7 and recovering said polypeptide.
- 9. A method for preventing or treating thromboembolic diseases or glomerulonephritis, which comprises administering to a patient in need thereof a disease-preventing or disease-treating effective amount of the polypeptide of claim 1.
- 10. An isolated and purified DNA sequence which codes for a polypeptide as defined in claim 2.
- 11. A recombinant DNA molecule comprising a DNA sequence as defined in claim 10 which is functionally connected to an expression control system which makes expression possible in suitable host systems.
- 12. A recombinant DNA molecule as defined in claim 11 wherein the expression control sequence is an E. coli promoter system, a promoter system of an E. coli bacteriophage, a yeast or fungus expression control sequence or another eukaryotic expression control sequence.
- 13. A host cell which contains at least one recombinant DNA molecule as defined in claim 12.
- 14. A host cell of claim 13, wherein said cell is a bacterial, yeast, fungal, animal or human cell.
- 15. A method for preventing or treating thromboembolic diseases or glomerulonephritis, which comprises administering to a patient in need thereof a disease-preventing or disease-treating effective amount of the polypeptide of claim 2.
- 16. The polypeptide of claim 1 wherein Xaa is selected from the group consisting of Gln, Asn, Ser, Thr, Gly, Asp, Glu, Lys, Arg and Pro.
- 17. The polypeptide of claim 1 wherein Xaa in positions 31, 135, 150 and 213 of the amino acid sequence represented by SEQ ID NO: 4 is selected from the group consisting of Phe, Tyr, Leu, Ile, Ala, Thr and Ser.
- 18. The polypeptide of claim 1 wherein Xaa is Asn or Gln.
- 19. The polypeptide of claim 18 wherein Xaa is Asn.
- 20. The polypeptide having an amino acid sequence set forth in SEQ ID NO: 3, wherein Xaa is selected from the group consisting of Asn, Gln, Ser, Thr, Gly, Asp, Lys, Arg and Pro.
- 21. The polypeptide of claim 20 wherein Xaa is Asn or Gln.
- 22. The polypeptide of claim 21 wherein Xaa is Asn.
- 23. The polypeptide having an amino acid sequence set forth in SEQ ID NO: 6, wherein Xaa is selected from the group consisting of Asn, Gln, Gly, Asp, Lys, Arg and Pro.
- 24. The polypeptide of claim 23 wherein Xaa is Asn or Gln.
- 25. The polypeptide of claim 24 wherein Xaa is Asn.
- 26. The polypeptide of claim 1 wherein Xaa in position 20 of the amino acid sequence represented by SEQ ID NO: 6 is selected from the group consisting of Phe, Tyr, Leu, Ile, Ala, Thr and Ser.
- 27. An isolated and purified deoxyribonucleic acid coding for a polypeptide as defined in claim 1 comprising at least one base sequence selected from the group consisting of a first base sequence represented by SEQ ID NO: 8 and a second base sequence complementary to said first base sequence.
- 28. An isolated and purified deoxyribonucleic acid coding for a polypeptide as defined in claim 1 comprising at least one base sequence selected from the group consisting of a first base sequence represented by SEQ ID NO: 11 and a second base sequence complementary to said first base sequence.
Priority Claims (1)
Number |
Date |
Country |
Kind |
40 23 699.4 |
Jul 1990 |
DEX |
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Parent Case Info
This application is a continuation of application Ser. No. 08/361,705, filed on Dec. 22, 1994 which is a continuation of application Ser. No. 07/966,040, filed as PCT/EP91/01361, Jul. 19, 1991, published as WO92/01794, Feb. 6, 1992, both now abandoned.
US Referenced Citations (3)
Number |
Name |
Date |
Kind |
4027012 |
Antonini |
May 1977 |
|
4585653 |
Williams |
Apr 1986 |
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4610879 |
Markland, Jr. et al. |
Sep 1986 |
|
Foreign Referenced Citations (2)
Number |
Date |
Country |
0 395 375 |
Oct 1990 |
EPX |
WO9006362 |
Jun 1990 |
WOX |
Non-Patent Literature Citations (1)
Entry |
Biochem. J. (1973) 131, pp. 799-807, Hatton, "Studies on the Coagulant Enzyme from Agkistrodon rhodostoma Venom". |
Continuations (2)
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Number |
Date |
Country |
Parent |
361705 |
Dec 1994 |
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Parent |
966040 |
Dec 1992 |
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