Determination of the kinetic constants of tissue factor/factor VII/factor VIIA and antithrombin/heparin using surface...

Association of factor VIIa (FVIIa) with tissue factor (TF) is generally believed to be a critical step in the initiation of blood coagulation. In this study the rate constants for the complex formation and dissociation between FVII/FVIIa and TF were studied in real time using surface plasmon resonance, the BIAcore technique. Employing this technique ka and kd were determined and yielded a KD of 1 nM for FVII and 0.5 nM for FVII, respectively. The association and dissociation between antithrombin (AT) and the FVIIa/TF complex was also studied. In the presence of heparin, AT was bound to the FVIIa/TF complex in a dose-dependent manner with ka of 2 x 10(3) M-1 s-1. The binding of AT to FVIIa/TF increased the dissociation of FVIIa from TF about 20-fold.