毕赤酵母重组Annexin32的分子特征及其对血凝与血栓形成的影响  

Characteristics of the Recombinant Annexin 32 in Pichia pastoris and its Effects on Coagulation Time and Thrombosis

作　　者：杨湘越[1] 兰小鹏[1] 颜宏利[2] 张毅[2] 朱忠勇[1] 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,福州350025 [2]第二军医大学医学遗传学教研室,上海200433

出　　处：《中国生物工程杂志》2004年第11期33-37,共5页China Biotechnology

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

摘　　要：采用SDS PAGE、等电聚焦、脱磷酸反应、蛋白质N端氨基酸测序、KPTT及大鼠下腔静脉血栓形成观察 ,发现毕赤酵母重组Annexin32的分子特征较原核产物有了明显变化。有主、次 2条带 ,主带的分子量比原核产物大 ,等电点较理论值低 ,已被磷酸化 ,2条带的N端均携带了来自载体的 4个氨基酸 ,可能对其与活化血小板表面磷脂的结合有所下调 ,导致血凝与血栓形成受到一定的影响 ,但仍具有明显的抗凝及抑制血栓形成作用。The recombinant annexin32 in Pichia pastoris was assayed by SDS-PAGE,IEF,dephosphorization,sequencing of amino-N-terminal,KPTT and venous thrombosis in inferior vena cava of rats.Characteristics of the recombinant annexin32 in Pichia pastoris varied from that of in E.coli. The r-annexin32 showed two bands.MW of the majar band was larger than that of r-annexin32 in E.coli.Its pI was lower than the theorial pI value of annexin32 because of phosphorization.The amino-N-terminus of two bands carried 4 aminoes from the yeast expression plasmid.These changes might decrease affinity with phospholipid-binding to activated pellet and affect coagulation time and thrombosis.However,the r-annexin32 in Pichia pastoris still had strong anticoagulation and antithrombosis.;

关 键 词：血栓形成 血凝 原核 分子特征 Annexin32 KPTT 活化血小板 毕赤酵母 等电点 等电聚焦 

分 类 号：Q786[生物学—分子生物学] R446[医药卫生—诊断学]