Purification and Properties of an Abnormal Blood Coagulation Factor IX (Factor IX_Bm)/Kinetics of Its Inhibition of Factor X Activation by Factor VII and Bovine Tissue Factor

Summary

An abnormal blood coagulation factor IX has been isolated from the blood of a hemophilia B patient with a variant of the disease (hemophilia B_m) characterized by a normal concentration of factor IX antigen, negligible factor IX coagulant activity, and a prolonged prothrombin time with bovine tissue factor. The isolated protein (factor IX_Bm) had the same apparent molecular weight as normal factor IX (55,000) and the same mobility on two dimensional immunoelectrophoresis as normal factor IX. Factor IX_Bm underwent limited proteolysis induced by activated factor XI, in the presence of Ca²⁺ ions, or induced by the reaction product of tissue factor, factor VII and Ca²⁺ ions. A timecourse study showed that activated factor XI cleaved factor IX_Bm and factor IX at similar rates. However, in contrast to normal factor IX, the limited protelysis of factor IX_Bm did not generate procoagulant activity.

In kinetic experiments purified factor IX_Bm behaved like a competitive inhibitor (K_i of 0.017 μM) of the activation of factor X by bovine tissue factor and factor VII. Normal factor IX was also found to inhibit the reaction but required a four-fold higher concentration to achieve the same inhibitory effects as factor IX_Bm.

Keywords

Factor IX_Bm - Activation of factor IX_Bm - Inhibition kinetics

• References

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