Macromolecular specificity determinants on thrombin for fibrinogen and thrombomodulin.
The endothelial cell surface membrane protein thrombomodulin binds thrombin with high affinity and acts as both a cofactor for protein C activation and an inhibitor of fibrinogen hydrolysis. We have previously shown that bovine thrombomodulin is a competitive inhibitor of fibrinogen binding to thrombin but has no effect on thrombin activity toward tripeptide substrates or antithrombin III. Hence, thrombomodulin and fibrinogen may share macromolecular specificity sites on thrombin which are distinct from the active site. In this investigation, we have studied the interaction of thrombin-thrombomodulin with fibrinogen and various thrombin derivatives. We show that fibrinogen is a competitive inhibitor of thrombomodulin binding to thrombin, with a Kis = 10 microM. Thrombin derivatives (bovine (pyridoxal phosphate)4-thrombin and human thrombin Quick I), which bind fibrinogen with much reduced affinity, are shown to also interact with thrombomodulin with greatly reduced affinity. These results are consistent with the hypothesis that thrombomodulin and fibrinogen share macromolecular specificity sites on thrombin.
Jakubowski
H V
HV
Department of Biochemistry, University of Iowa, Iowa City 52242.
Owen
W G
WG
D001665:Binding Sites; D001667:Binding, Competitive; D005340:Fibrinogen / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D011486:Protein C / Q000037:antagonists & inhibitors; D011487:Protein Conformation; D011732:Pyridoxal Phosphate / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism* / Q000494:pharmacology; D018179:Receptors, Thrombin; D013917:Thrombin / Q000378:metabolism*
D046911:Macromolecular Substances; D011486:Protein C; D011956:Receptors, Cell Surface; D018179:Receptors, Thrombin; D011732:Pyridoxal Phosphate; D005340:Fibrinogen; D013917:Thrombin; C060096:tetra(pyridoxal phosphate)thrombin; C027648:thrombin Quick