fibrinopeptide-b and Hemophilia-A

The rate of conversion of fibrinogen (Fg) to the insoluble product fibrin (Fn) is a key factor in hemostasis. We have developed methods to quantitate fibrinopeptides (FPs) and soluble and insoluble Fg/Fn products during the tissue factor induced clotting of whole blood. Significant FPA generation (>50%) occurs prior to visible clotting (4 +/- 0.2 min) coincident with factor XIII activation. At this time Fg is mostly in solution along with high molecular weight cross-linked products. Cross-linking of gamma-chains is virtually complete (5 min) prior to the release of FPB, a process that does not occur until after clot formation. FPB is detected still attached to the beta-chain throughout the time course demonstrating release of only low levels of FPB from the clot. After release of FPB a carboxypeptidase-B-like enzyme removes the carboxyl-terminal arginine resulting exclusively in des-Arg FPB by the 20-min time point. This process is inhibited by epsilon-aminocaproic acid. These results demonstrate that transglutaminase and carboxypeptidase enzymes are activated simultaneously with Fn formation. The initial clot is a composite of Fn I and Fg already displaying gamma-gamma cross-linking prior to the formation of Fn II with Bbeta-chain remaining mostly intact followed by the selective degradation of FPB to des-Arg FPB.

Topics: Adolescent; Adult; Amino Acid Sequence; Aminocaproic Acid; Blood Coagulation; Carboxypeptidase B; Carboxypeptidases; Enzyme Activation; Factor XIII; Fibrin; Fibrinogen; Fibrinopeptide A; Fibrinopeptide B; Hemophilia A; Humans; Models, Biological; Molecular Sequence Data; Protease Inhibitors; Protein Conformation; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Thromboplastin; Transglutaminases