摘要
目的 体外组装包含人C4分子的补体经典激活途径C3转化酶 ,并对其转化酶活性及衰变特性进行观察。方法 利用豚鼠血清功能纯C1、C2及溶血中间体EAC4 hu体外组装经典途径C3转化酶 ,观察不同C1、C2用量及孵育温度对C3转化酶形成和自发性衰变的影响 ,以及人红细胞膜抽提蛋白对C3转化酶衰变的影响。结果 高剂量和低剂量的C1均会影响C3转化酶的形成 ,增加C2用量可增加C3转化酶的形成数量 ,C3转化酶的自发性衰变随孵育温度的升高而加速 ,人红细胞膜抽提蛋白可抑制C3转化酶的自发性衰变过程。结论 C1、C2用量及孵育温度是影响C3转化酶形成和自发性衰变的主要因素 ,体外组装的补体经典激活途径C3转化酶可应用于相关补体调控蛋白的活性检测。
Objective To prepare classical pathway C3 convertase of complement system in vitro and to observe its activity. Methods Classical pathway C3 convertase of complement system was generated in vitro by functionally purified guinea pig C1, C2 and intermediate EAC4 hu . Effects of gp C1 or gp C2 concentration and incubation temperature on the formation and decay of prepared C3 convertase were then observed. The assay of decay accelerating (DA) activity of the classical pathway C3 convertase was then established and used to detect DA activity in the extracts of human erythrocyte membranes. Results Both high concentration and low concentration of gp C1 decreased the amount of C3 convertase generated, while higher gp C2 concentration increased the C3 convertase. The decay of C3 convertase was accelerated when it was incubated in high temperature. Extracts of human erythrocyte membranes inhibited the decay of C3 convertase. Conclusions gp C1 or gp C2 concentration and incubation temperature are key factors in the formation and decay of the classical pathway C3 convertase. C3 convertase prepared by this method can be used to detect DA activity of related complement regulatory proteins.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2002年第6期599-602,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(3 0 0 80 0 3 2 )
重庆市科委重点攻关计划项目
