Extrinsic activation of human blood coagulation factors IX and X.
We studied activation of human coagulation factors IX and X by factor VIIa in the presence of calcium ions, phospholipid (phosphatidylserine/phosphatidylcholine, 50/50, mol/mol) and purified tissue factor apoprotein. Activation of factor IX and factor X was found to occur without a measurable lag-phase and hence initial rates of factor IXa and factor Xa formation could be determined. Like previously observed for the activation of factor X, the activation of factor IX was saturable with respect to factor VIIa, tissue factor apoprotein and phospholipid. The results suggested that in the presence of a Ca2+ ions the same ternary complex of factor VIIa-tissue factor apoprotein-phospholipid is responsible for the activation of factor IX and factor X. Both the apparent Km of 22 nM-factor IX and the apparent Kcat of 28 min-1 were about 3-fold lower than the corresponding parameters of factor X activation by this complex. Hence, the catalytic efficiency (Kcat/Km) of factor IX and factor X activation was about equal. However, the two substrates inhibited the activation of each other by competition for the same catalytic sites. The apparent Kinh of factor IX for inhibition of extrinsic factor X activation is 30 nM. The apparent Kinh of factor X for inhibition of extrinsic factor IX activation is 116 nM. From these kinetic data it was calculated that at plasma concentration of factors IX and X, the rate of extrinsic factor IX activation would be half the rate of factor X activation.(ABSTRACT TRUNCATED AT 250 WORDS)
Bom
V J
VJ
Division of Haemostasis and Thrombosis, University Hospital Groningen, The Netherlands.
Reinalda-Poot
J H
JH
Cupers
R
R
Bertina
R M
RM
D001059:Apoproteins / Q000378:metabolism; D001667:Binding, Competitive; D001779:Blood Coagulation Factors / Q000302:isolation & purification; D015949:Factor IXa / Q000096:biosynthesis*; D015942:Factor VIIa / Q000378:metabolism; D015951:Factor Xa / Q000096:biosynthesis*; D006801:Humans; D015592:Immunoradiometric Assay; D007457:Iodine Radioisotopes; D007700:Kinetics; D010713:Phosphatidylcholines / Q000097:blood; D010718:Phosphatidylserines / Q000097:blood; D013053:Spectrophotometry
D001059:Apoproteins; D001779:Blood Coagulation Factors; D007457:Iodine Radioisotopes; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D015942:Factor VIIa; D015949:Factor IXa; D015951:Factor Xa