SummaryTissue thromboplastin apoprotein was partially purified from human brain. The apoprotein was recombined with mixed phospholipids to yield active thromboplastin. The recombined thromboplastin induced proteolytic activation of isolated human factor IX in the presence of factor VII and Ca2+. The clotting times of various deficient plasmas were determined as a function of apoprotein concentration, keeping the phospholipid concentration constant. The clotting times of a factor XII-deficient plasma were the same as those of a factor XII/factor IX-deficient plasma, except at very low apoprotein concentrations. However, under those conditions the difference in clotting times was independent of the presence of anti-factor VII serum. Similar observations were made for factor XI-deficient plasma in comparison with factor XI/factor IX-deficient plasma. These results indicate that activation of factor IX by factor VII/tissue thromboplastin does not significantly contribute to plasma coagulation.
This study is concerned with the question whether activation of factor IX by factor VII - tissue thromboplastin contributes to the rate of plasma coagulation. The protein component of tissue factor was partially purified from human brain. Its molecular weight as deduced from SDS - polyacrylamide gel electrophoresis was about 48,000. Reconstitution of thromboplastin activity was obtained by mixing apoprotein and phospholipids in the presence of Triton X-100 and subsequent removal of Triton by adsorption to Biobeads SM-2. Reconstituted tissue factor greatly accelerated the activation of factor IX by isolated factor VII in the presence of calcium ions. In a contact free system (plasma from a patient with congenital factor XII deficiency; factor XII<0.001 Unit/ml) plasma coagulation times (tc) were determined as a function of apoprotein concentration (at constant phospholipid) both in the presence and absence of factor IX. At high apoprotein concentration tc showed to be independent of factor IX, whereas at low apoprotein concentration the removal of factor IX resulted in a 2 - 3 fold increase of tc. The involvement of the tissue factor - factor VII complex in this phenomenon was evaluated using a specific anti-factor VII serum. The results indicate that activation of factor IX by factor VII - tissue thromboplastin does not significantly contribute to the rate of plasma coagulation.
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