Kinetics of the inhibition of tissue factor-factor VIIa by tissue factor pathway inhibitor.
Tissue factor-factor VIIa catalysed activation of factor IX is inhibited by the complex of tissue factor pathway inhibitor (TFPI) and factor Xa. At present, no information is available as to what extent the kinetics of complex formation between TFPI and factor Xa during factor X activation contribute to the overall rate of inactivation of the factor X converting complex. We have determined the kinetic parameters of the individual reactions, i.e. factor X activation, formation of the TFPI-factor Xa complex, and inactivation of tissue factor-factor VIIa by the TFPI-factor Xa complex. We modelled the overall reaction by assuming a two-step reaction: factor Xa generated by tissue factor-factor VIIa forms a reversible complex with TFPI and in the second step this complex forms a reversible quaternary complex with tissue factor-factor VIIa. The validity of the model was demonstrated by analysis of factor Xa generation curves in the presence of TFPI. Independently determined constants for factor X activation (kcat = 12 s-1, Km = 70 nM) and inhibition of tissue factor-factor VIIa by TFPI-factor Xa complex (rate constant of inhibition of 1.1 x 10(8) M-1S-1) were used. The association rate constant of the formation of the TFPI-factor Xa complex was estimated by fitting the model to the data.(ABSTRACT TRUNCATED AT 250 WORDS)
Lindhout
T
T
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Franssen
J
J
Willems
G
G
D000595:Amino Acid Sequence; D000925:Anticoagulants / Q000494:pharmacology*; D003198:Computer Simulation; D015942:Factor VIIa / Q000037:antagonists & inhibitors* / Q000737:chemistry; D065427:Factor Xa Inhibitors; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008074:Lipoproteins / Q000494:pharmacology*; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D013925:Thromboplastin / Q000037:antagonists & inhibitors* / Q000737:chemistry
D000925:Anticoagulants; D065427:Factor Xa Inhibitors; D005343:Fibrinolytic Agents; D008074:Lipoproteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C051928:lipoprotein-associated coagulation inhibitor; C081249:tissue factor pathway inhibitor (1-161); D013925:Thromboplastin; D015942:Factor VIIa