Modified vitamin K-dependent polypeptides
The invention provides vitamin K-dependent polypeptides with enhanced membrane binding affinity. These polypeptides can be used to modulate clot formation in mammals. Methods of modulating clot formation in mammals are also described.
1. A composition comprising an anticoagulant agent and a protein C polypeptide or activated protein C polypeptide wherein said polypeptide comprises a modified GLA domain, said modified GLA domain comprising the amino acid sequence of SEQ ID NO: 1 with one, two, three, four, or five amino acid substitutions at positions selected from the group consisting of residues 10, 11, 28, 32, and 33.
2. The composition of claim 1 , wherein said anticoagulant agent is aspirin, warfarin, or heparin.
3. The composition of claim 1 , wherein said anticoagulant agent is aspirin.
4. The composition of claim 1 , wherein said one amino acid substitution is at residue 10.
5. The composition of claim 1 , wherein said one amino acid substitution is at residue 11.
6. The composition of claim 1 , wherein said one amino acid substitution is at residue 28.
7. The composition of claim 1 , wherein said one amino acid substitution is at residue 33.
8. The composition of claim 1 , wherein said modified GLA domain comprises the amino acid sequence of SEQ ID NO:1 with three amino acid substitutions at residues 11, 32, and 33.
9. The composition of claim 8 , wherein residue 21 of SEQ ID NO:1 is glutamic acid and residue 33 of SEQ ID NO:1 is aspartic acid.
10. The composition of claim 9 , wherein residue 11 of SEQ ID NO:1 is glycine.
11. The composition of claim 1 , wherein said modified GLA domain comprises the amino acid sequence of SEQ ID NO:1 with two amino acid substitutions at residues 32 and 33.
12. The composition of claim 11 , wherein residue 32 of SEQ ID NO:1 is glutamic acid and residue 33 of SEQ ID NO:1 is aspartic acid.