Amino acid modified polypeptides

• US 20050196830A1
• Filed: 03/22/2002
• Published: 09/08/2005
• Est. Priority Date: 06/03/1996
• Status: Abandoned Application

• Current Assignee: Douglas D. Buechter, Elliott A. Gruskin, Guanghui Zhang, Kevin Connolly
• Original Assignee: Douglas D. Buechter, Elliott A. Gruskin, Guanghui Zhang, Kevin Connolly
• Inventors: Zhang, Guanghui, Gruskin, Elliott A., Connolly, Kevin, Buechter, Douglas D.
• Application Number:
  US10/104,793
• Publication Number:
  US 20050196830A1
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: C07K 14/4725 Proteoglycans, e.g. aggreccan
• CPC Class Codes: C07K 14/495 Transforming growth factor [TGF]
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C07K 14/75 Fibrinogen
• 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: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione

Abstract

Methods are disclosed for simplified recombinant production of fibrillar collagens. DNAs encoding fibrillar collagen monomers lacking the N propeptide, the C propeptide, or both propeptides are introduced into recombinant host cells and expressed. Trimeric collagen is recovered from the recombinant host cells.

Claims

1. Purified human collagen or fragment thereof produced by a prokaryotic cell, the purified human collagen or fragment thereof being capable of providing a self aggregate, wherein the purified human collagen or fragment thereof has incorporated therein at least one amino acid which has not undergone post translational enzymatic modification selected from the group consisting of trans-4-hydroxyproline and 3-hydroxyproline, and the purified human collagen or fragment thereof having the at least one amino acid self aggregates to form an extracellular matrix.

2. Human collagen or fragment thereof produced by a prokaryotic cell according to claim 1 wherein the human collagen or fragment thereof is encoded for by nucleic acid having the sequence shown in SEQ. ID. NO. 19.

3. Human collagen or fragment thereof produced by a prokaryotic cell according to claim 1 wherein the human collagen or fragment thereof is encoded for by nucleic acid having the sequence shown in SEQ. ID. NO. 39.

4. Human collagen or fragment thereof produced by a prokaryotic cell according to claim 1 wherein the human collagen or fragment thereof is encoded for by nucleic acid having the sequence shown in SEQ. ID. NO. 43.

5. Human collagen or fragment thereof produced by a prokaryotic cell according to claim 1 wherein the human collagen or fragment thereof is encoded for by nucleic acid having the sequence shown in SEQ. ID. NO. 45.

6. Purified human collagen or fragment thereof produced by a prokaryotic cell according to claim 1 wherein the purified collagen or fragment thereof is encoded for by nucleic acid having the sequence shown in SEQ. ID. NO. 31.

7. Purified human collagen or fragment thereof produced by a prokaryotic cell, the purified human collagen or fragment thereof being capable of providing a self aggregate, wherein the purified human collagen or fragment thereof has incorporated therein at least one amino acid comprising 3-hydroxyproline which has not undergone post translational enzymatic modification, and the purified human collagen or fragment thereof having the at least one amino acid self aggregates to form an extracellular matrix.

8. Human collagen or a fragment thereof produced by a prokaryotic cell wherein all naturally occurring sites which contain proline or hydroxyproline are occupied by trans-4-hydroxyproline or 3-hydroxyproline, wherein the collagen or fragment thereof have the property of self-aggregation.

9. Human collagen or a fragment thereof according to claim 8, wherein the human collagen is human type I (α

₂) collagen.

10. Human collagen or a fragment thereof according to claim 9, wherein the collagen or fragment thereof is encoded by nucleic acid having the sequence shown in SEQ.ID.NO. 31.