Kinetic aspects of release of fibrinopeptides AP and AY by human alpha-thrombin

The time-dependent release by human alpha-thrombin of the physiological variants of fibrinopeptide A, i.e. fibrinopeptide A-3-phosphate (FPAP) and des-Ala-fibrinopeptide A (FPAY), has been measured in order to study the kinetic pathway for their hydrolysis. The best-fit kinetic model for the release of FPAP and FPAY is consistent with a simple pseudo first-order reaction, as observed with FPA. These findings indicate that FPAP and FPAY are also released from intact fibrinogen before release of FPB. The values of the specificity constants, i.e. k(at)/Km, for the enzymatic reaction between thrombin and the various alpha-chains showed that AP alpha- and AY alpha-chain are hydrolyzed with a 0.2-and 0.4-fold higher specificity constant respectively than that of A alpha-chain. Such minor differences observed with the various chains suggest that the 1-3 NH2-terminal residues of the chain do not significantly contribute to the catalytic efficiency of thrombin toward the alpha-chains of fibrinogen.