Effects of Neutral Proteases from Human Leukocytes on Structure and Biological Properties of Human Factor VIII
Effects of Neutral Proteases from Human Leukocytes on Structure and Biological Properties of Human Factor VIII
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人白细胞中性蛋白酶对人因子 VIII 结构和生物学特性的影响
DOI：
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发表时间：
1980
影响因子：
6.7
通讯作者：
E. Wojtecka
中科院分区：
文献类型：
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作者：
M. Kopeć;K. Bykowska;S. Lopaciuk;M. Jeleńska;J. Kaczanowska;I. Sopata;E. Wojtecka
人因子VIII从低温沉淀中纯化，与四种人血液白细胞中性蛋白酶，即弹性酶样蛋白酶（ELP）、凝乳胰蛋白酶样蛋白酶（CLP）、胶原酶和明胶酶孵育24小时。电泳模式显示了ELP和CLP降解因子VIII及其23万分子量亚基的可重复序列。中间产物相似，但ELP和CLP穷举蛋白水解产物差异明显。在检测到明显的电泳变化之前，因子VIII的促凝活性被ELP和CLP迅速而完全地破坏。在销毁之前，没有观察到这种活动增加。冯氏血友病因子（立斯托霉素辅助因子）活性对ELP和CLP的抗性更强，与高度聚集形式的因子VIII的降解大致相关。ELP和CLP使劳雷尔反应抗原显著进行性升高。正常人血浆对ELP和CLP有较强的抑制作用。大剂量的这些酶（每毫升300微克）在血浆培养基中只产生因子VIII促凝活性的适度下降。胶原酶和明胶酶既不能降解因子VIII，也不能改变其生物学特性。
Summary Human factor VIII was purified from cryoprecipitate and incubated for up to 24 hours with four neutral proteases of human blood leukocytes, namely, with elastase-like protease (ELP), chymotrypsin-like protease (CLP), collagenase and gelatinase. Electrophoretic patterns showed a reproducible sequence of degradation of factor VIII and of its 230,000 molecular weight subunit by ELP and CLP. Intermediate products were similar but those resulting from exhaustive proteolysis by ELP and CLP differed distinctly from each other. Procoagulant activity of factor VIII was rapidly and completely destroyed by ELP and CLP before visible electrophoretic changes would be detected. No increase in this activity was observed prior to its destruction. Von Willebrand factor (ristocetin cofactor) activity was considerably more resistant to ELP and CLP and declined in rough relation to degradation of highly aggregated forms of factor VIII. ELP and CLP produced a pronounced progressive increase in the Laurell reaction antigen. Normal human plasma showed a high potency to inhibit ELP and CLP. Large doses of these enzymes (300 ug per ml) produced in the plasma medium only a moderate fall in factor VIII procoagulant activity. Collagenase and gelatinase did neither degrade factor VIII nor change its biological properties.