Active Site-labeled Prothrombin Inhibits Prothrombinase in Vitro and Thrombosis in Vivo

Heather K. Kroh, Peter Panizzi, Svetlana Tchaikovski, T. Regan Baird, Nancy Wei, Sriram Krishnaswam, Guido Tans, Jan Rosing, Bruce Furie, Barbara C. Furie & Paul E. Bock
Mouse and human prothrombin (ProT) active site specifically labeled with D-Phe-Pro-Arg-CH2Cl (FPR-ProT) inhibited tissue factor-initiated thrombin generation in platelet-rich and platelet-poor mouse and human plasmas. FPR-prethrombin 1 (Pre 1), fragment 1 (F1), fragment 1.2 (F1.2), and FPR-thrombin produced no significant inhibition, demonstrating the requirement for all three ProT domains. Kinetics of inhibition of ProT activation by the inactive ProTS195A mutant were compatible with competitive inhibition as an alternate nonproductive substrate, although FPR-ProT deviated from this...
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