Peptides comprising repetitive units of amino acids and DNA sequences encoding the same

• US 6,018,030 A
• Filed: 06/07/1995
• Issued: 01/25/2000
• Est. Priority Date: 11/04/1986
• Status: Expired due to Term

• Current Assignee: Syntro Corp. (Mallinckrodt plc), Protein Polymer Technologies, Inc., Spine Wave Incorporated
• Original Assignee: Protein Polymer Technologies, Inc.
• Inventors: Ferrari, Franco A., Cappello, Joseph, Richardson, Charles, Chambers, James, Pollock, Thomas J., Causey, Stuart, Crissman, John W.
• Primary Examiner(s): Degen, Nancy
• Assistant Examiner(s): SANDALS, WILLIAM O
• Application Number:
  US08/482,085
• Time in Patent Office: 1,693 Days
• Field of Search: 435/6, 435/69.1, 435/240.1, 435/325, 436/86, 530/353, 536/23.1, 536/23.5
• US Class Current: 530/353
• CPC Class Codes: C07K 14/43586 from silkworms
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/61 containing an enzyme fusion for detection (lacZ, luciferase)
• CPC Class Codes: C12N 1/06 Lysis of microorganisms
• CPC Class Codes: C12N 15/11 DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/66 General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 15/75 for Bacillus

Abstract

Polypeptides comprising repetitive units of amino acids, as well as synthetic genes encoding the subject polypeptides are provided. The subject polypeptides are characterized by comprising repetitive units of amino acids, where the repetitive units are present in naturally occurring proteins, particularly naturally occurring structural proteins. The subject polypeptides find use in a variety of applications, such as structural components of prosthetic devices, synthetic fibers, and the like.

Claims

1. A recombinantly produced protein of from 30 kDal to 250 kDal comprising at least 50 number percent of amino acids of at least one naturally occurring repeating unit of a naturally occurring structural protein, wherein said repeating units are from 4 to 20 amino acids in length and wherein each of said repeating units which comprise said at least 50 number percent of the amino acids of said protein are tandemly repeated at least once in the protein.

2. The recombinantly produced protein according to claim 1, wherein said repeating units are of from 4 to 15 amino acids.

3. The recombinantly produced protein according to claim 1, wherein said repeating units are of from 4 to 9 amino acids.

4. A formed object comprising the protein according to claim 1.

5. The recombinantly produced protein according to claim 1, wherein said repeating units are present in at least 2 blocks of at least 2 repeating units per block.

6. The recombinantly produced protein according to claim 5, wherein said protein comprises an intervening group of from 1 to 50 amino acids between at least 2 of said blocks of repeating units.

7. A DNA sequence encoding the protein according to claim 1.

8. A recombinantly produced protein of from 30 kDal to 250 kDal comprising at least 50 number percent of amino acids of at least one naturally occurring repeating unit of a naturally occurring structural protein selected from the group consisting of elastin, silk, and keratin and of from 4 to 20 amino acids in length and wherein each of said repeating units which comprise said at least 50 number percent of the amino acids of said protein are tandemly repeated at least once in the protein.

9. The recombinantly produced protein according to claim 8, wherein said at least one repeating unit is selected from the group consisting of SGAGAG (SEQ ID NO:

21) and GAGAGS (SEQ ID NC;

41).

10. The recombinantly produced protein according to claim 8, wherein said at least one repeating unit is selected from the group consisting of GVGVP (SEQ ID NO:

23), VPGVG (SEQ ID NO;

24) and APGVGV (SEQ ID NO;

25).

11. The recombinantly produced protein according to claim 8, wherein said at least one repeating unit is K-L-(1)-L-A-E-A (SEQ ID NO:

105), wherein 1 is a basic or acidic amino acid, particularly K or E and the repeating units alternate as to whether 1 is a basic or acidic amino acid.

12. A formed object comprising the protein according to claim 8.

13. The recombinantly produced protein according to claim 8, wherein said repeating units are present in at least 2 blocks of at least 2 repeating units per block.

14. The recombinantly produced protein according to claim 13, wherein said protein comprises an intervening group of from 1 to 50 amino acids between at least 2 of said blocks of repeating units.

15. A DNA sequence encoding the protein according to claim 8.

19. A cell comprising the DNA sequence according to any one of claims 7, 15 or 18.

16. A recombinantly produced protein of from 30kDal to 250kDal comprising at least 50 number percent of amino acids of at least one naturally occurring repeating unit of the amino acid sequence Gly-α

-β

, wherein α and

β

are any amino acid and are selected so that the encoded protein comprises from about 10 to 45 number percent proline.

17. A formed object comprising the protein according to claim 16.

18. A DNA sequence encoding the protein according to claim 16.