乙型肝炎病毒PreS1蛋白与初期多肽相关复合体α亚单位特异性结合的研究  

Interaction between hepatitis B virus PreS1 protein and nascent-polypeptide-associated complex alpha polypeptide

作　　者：李丹[1] 丁健[2] 林纳[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化研究室,福州350001 [2]福建医科大学附属第一医院消化内科,福州350001

出　　处：《第三军医大学学报》2009年第23期2315-2318,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30770979);福建省自然科学基金(C0310018)~~

摘　　要：目的探讨乙型肝炎病毒(HBV)PreS1蛋白与人类初期多肽相关复合体α亚单位(nascent-polypeptide-asso-ciated complex alpha polypeptide,NACA)之间的结合作用。方法采用交合实验验证PreS1蛋白和NACA在酵母细胞内的结合作用,利用离体结合实验证实二者在体外的结合作用,免疫共沉淀实验证实二者在哺乳动物细胞中的特异性结合。结果携带NACA基因的酵母同携带preS1基因的酵母交合后发生反应,LacZ活性检测菌落呈现蓝色,离体结合实验Western印迹中出现特异性结合活性反应条带,提示PreS1蛋白和NACA在体外存在直接相互作用,免疫共沉淀实验在13×103处出现蛋白条带,提示在哺乳动物细胞水平仍能检测到PreS1蛋白和NACA的特异结合。结论NACA与PreS1蛋白在酵母细胞和哺乳动物细胞中均存在特异性结合,推测NACA可能通过与PreS1蛋白的结合影响病毒黏附及反式激活等过程。Objective To explore the interaction between hepatitis B virus PreS1 protein and nascentpolypeptide-associated complex alpha polypeptide （NACA）. Methods Mating experiment and in vitro binding assays were carried out to verify the interaction between hepatitis B virus PreS1 protein and NACA in vivo and in vitro respectively in yeast cells. Coimmunoprecipitation was employed to demonstrate the binding reaction between the 2 proteins in mammalian cells. Results In mating experiment, the resulting diploids containing pAS2-preS1 and pACT2-NACA plasmids turned to blue in the X-gal assay. Western blot analysis displayed a positive reactive strap, which indicating PreS1 protein interacted directly with NACA in vitro binding assays. Moreover, interaction between the 2 proteins in mammalian cells was confirmed by coimmunoprecipitation, with a protein strap about 13 ×10^3 in Western blot. Conclusion Specific interaction exists between hepatitis B virus PreS1 protein and NACA in both prokaryotic and mammalian cells. NACA protein probably takes part in the adhesion and transactivation of HBV through the bindinz with PreSi.

关 键 词：肝炎病毒 乙型 PRES1蛋白 初期多肽相关复合体α亚单位 离体结合实验 免疫共沉淀实验 

分 类 号：R341[医药卫生—基础医学] R373.21