Giddings J C, Bloom A L
Br J Haematol. 1975 Feb;29(2):349-64. doi: 10.1111/j.1365-2141.1975.tb01829.x.
Purified coagulation factors and specific antibodies to factor V and factor X were used to investigate the action of thrombin on factor V and the mechanism by which thrombin-treated factor V influences prothrombin activation. The formation of a complex or complexes between phospholipid, factor V, factor Xa and calcium was demonstrated by column chromatography on Sephadex gel, and by immunological analysis of the column fractions including the use of solid-phase antibodies. Kinetic experiments demonstrated that generation of thrombin from purified prothrombin was accomplished by this complex. Pre-treatment of factor V with trace quantities of thrombin resulted in increased yield and rate of thrombin generation. It was shown that phospholipid became saturated when incubated with increasing concentrations of factor V and that the initial saturating concentration of the latter was reduced by pre-treatment with thrombin. The findings confirm that optimum conversion of prothrombin to thrombin is accomplished by a complex or complexes of phospholipid, factor V, factor Xa and calcium and it is suggested that thrombin plays an autocatalytic role in these reactions.
使用纯化的凝血因子以及针对因子V和因子X的特异性抗体，来研究凝血酶对因子V的作用，以及凝血酶处理后的因子V影响凝血酶原激活的机制。通过在葡聚糖凝胶上进行柱色谱分析，以及对包括使用固相抗体在内的柱级分进行免疫分析，证实了磷脂、因子V、因子Xa和钙之间形成了一种或多种复合物。动力学实验表明，由纯化的凝血酶原生成凝血酶是通过这种复合物实现的。用微量凝血酶预处理因子V可提高凝血酶生成的产量和速率。结果表明，当与浓度不断增加的因子V一起孵育时，磷脂会饱和，并且通过凝血酶预处理可降低后者的初始饱和浓度。这些发现证实，凝血酶原向凝血酶的最佳转化是通过磷脂、因子V、因子Xa和钙的一种或多种复合物来完成的，并且表明凝血酶在这些反应中起自催化作用。
