Designer collagens and use thereof
The present invention identified a recombinant synthetic collagen containing a triple helical backbone protein produced in a prokaryotic expression system where the protein contains at least one ‘inserted’ biologically active sequence(s).
1. A recombinant synthetic collagen comprising an isolated and purified protein produced in a prokaryotic expression system, wherein said protein comprises a collagen-like repeat GXYGX 1 Y 2 GX 2 Y 2 GXY SEQ ID NO: 13), wherein the amino acid in the X1 position is changed to L, R, or F, and/or the amino acid in the Y2 position is changed to R, K, or N, wherein the recombinant synthetic collagen forms a triple helical backbone.
2. The recombinant synthetic collagen of claim 1 , wherein said protein is derived from a Streptococcal protein.
3. The recombinant synthetic collagen of claim 1 , wherein said collagen is capable of binding to integrins α1β1 and α2β1 without hydroxyproline.
4. The recombinant synthetic collagen of claim 1 , wherein said biologically active sequences are shown in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, or SEQ ID NO: 12.
5. The recombinant synthetic collagen of claim 4 , wherein said collagen containing sequences shown in SEQ ID NO: 4, SEQ ID NO: 7 or SEQ ID NO: 10 support adherence of both α1β1 and α2β1 spreading of endothelial cells, fibroblasts, chondrocytes, and smooth muscle cells.
6. The recombinant synthetic collagen of claim 4 , wherein said collagen containing the sequence shown in SEQ ID NO: 11 selectively bind to integrin α1β1, but not to α2β1.
7. The recombinant synthetic collagen of claim 6 , wherein said collagen supports adherence of endothelial cells, fibroblasts, and chondrocytic cells, but does not support adherence of smooth muscle cells.
8. The recombinant synthetic collagen of claim 4 , wherein said collagen containing the sequence shown in SEQ ID NO: 10 supports adherence and spread of mesenchymal stem cells or adipocyte stem cells.
9. The recombinant synthetic collagen of claim 4 , wherein said collagen containing the sequence shown in SEQ ID NO: 11 supports adherence and spread of mesenchymal stem cells.
10. The recombinant synthetic collagen of claim 4 , wherein said collagen containing sequences shown in SEQ ID NO: 4, SEQ ID NO: 7, SEQ ID NO: 10, or SEQ ID NO: 11 do not aggregate platelets and are non-thrombogenic.
11. The recombinant synthetic collagen of claim 4 , wherein said collagen containing a GFPGER (SEQ ID NO: 10) sequence inhibits collagen-induced platelet aggregation.
12. The recombinant synthetic collagen of claim 4 , wherein said collagen containing the sequence shown in SEQ ID NO: 11 does not inhibit collagen-induced platelet aggregation.
13. The recombinant synthetic collagen of claim 4 , wherein said collagen contains one, two, three, four or five multiple cell binding motifs results in a density dependent increase in integrin affinity, cell binding, and cell migration.
14. The recombinant synthetic collagen of claim 13 , wherein said collagen contains one, two, three, four or five cell binding motifs shown in SEQ ID NO: 4.
15. The recombinant synthetic collagen of claim 1 , wherein said collagen is affixed to or linked in a chemical manner to a scaffold with intrinsic tensile properties.
16. The recombinant synthetic collagen of claim 15 , wherein said scaffold is selected from the group consisting of PEG-containing hydrogels, ECM components, and mesh materials.
17. The recombinant synthetic collagen of claim 1 , further comprising an insert selected from the group consisting of bone sialoprotein binding sequences, integrins a α10β1 and α11β1 binding sequences, and an extracellular matrix constituent.
18. The recombinant synthetic collagen of claim 1 , wherein said collagen is produced in a prokaryotic expression system deficient in post-translational modification.