新型重组膜联蛋白B1的特征及其抗血液凝固活性的比较  

Characteristics of A Noval Recombinant Annexin B1 in Pichia Pastoris and its Anti-Coagulation Activity

作　　者：杨湘越[1] 兰小鹏[1] 朱忠勇[1] 颜宏利[2] 张毅[2] 孙树汉[2] 

机构地区：[1]南京军区福州总医院全军医学检验中心(福建医科大学福总临床医学院),福州350025 [2]第二军医大学医学遗传学教研室,上海200433

出　　处：《福建医科大学学报》2005年第1期1-4,共4页Journal of Fujian Medical University

基　　金：国家 8 63计划资助项目 (2 0 0 1AA2 13 111)

摘　　要：目的　研究毕赤酵母重组膜联蛋白 B1的特征变化 ,比较其和原核表达产物对血细胞凝集与血栓形成过程的影响。　方法　对毕赤酵母重组膜联蛋白 B1行 SDS- PAGE,等电聚焦 ,脱磷酸反应 ,蛋白质 N端氨基酸测序 ,白陶土部分凝血活酶时间 (KPTT)测定及大鼠下腔静脉血栓形成观察。　结果　毕赤酵母重组膜联蛋白 B1显示两条电泳带 ,其中一条带的相对分子质量比原核表达产物大 ,已被磷酸化 ,等电点较理论值低 ,两条带的 N端均携带了来自载体的 4个氨基酸 ,毕赤酵母重组膜联蛋白 B1和大肠杆菌谷胱甘肽转移酶融合膜联蛋白 B1具有明显的抗凝及抑制血栓形成作用。　结论　虽然毕赤酵母重组膜联蛋白 B1的分子特征较原核产物有了明显变化 ,但其抗凝及抑制血栓功能基本未受影响。Objective To study characteristics of the recombinant annexin B1 in Pichia pastoris and its effect on anti-coagulation activity. Methods The characteristic of recombinant annexin B1 was assayed with SDS-PAGE, isoelectric focusing(IEF), dephosphorization, sequencing of amino-N-terminal, kaolin partial thromboplastin time(KPTT) and venous thrombosis in inferior vena cava of rats. Results The expression product of the recombinant annexin B1 in Pichia pastoris showed two bands: 40 kD band was larger than that of recombinant annexin B1 in E.coli, pI was lower than the theorial pI value of annexin B1 because of phosphorization. N-terminus of two bands contained 4 amino acids from the plasmid. KPTT and venous thrombosis in inferior vena cava of rats revealed that the recombinant annexin B1 in Pichia pastoris had significant anticoagulatant and antithrombotic effects corrected with glutathione transferase(GST) combinated annexin B1 in E.coli. Conclusion Although characteristics of the recombinant annexin B1 in Pichia pastoris varied from that of in E.coli, its anticoagulation and antithrombosis were sustained.

关 键 词：膜联蛋白 原核表达产物 血液凝固 抗凝 部分凝血活酶时间 KPTT 白陶土 毕赤酵母 等电点 电泳 

分 类 号：R341.3[医药卫生—基础医学]