HeLa细胞膜上日本脑炎病毒受体候选分子的初步鉴定  

A preliminary identification of protein candidate of JEV receptor on HeLa cell membrane

作　　者：王媛[1] 时莹[1] 寇甜甜 雷迎峰[1] 丁天兵[1] 

机构地区：[1]第四军医大学基础部微生物学教研室,陕西西安710032

出　　处：《热带医学杂志》2016年第1期18-21,27,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(31100123;30872216)

摘　　要：目的寻找HeLa细胞膜上可与日本脑炎病毒(JEV)结合的分子。方法采用免疫共沉淀(Co-IP)技术,分离HeLa细胞膜上可与JEV特异结合的分子,对SDS-PAGE特异条带进行质谱分析,并经Western blotting和酶联免疫吸附试验(ELISA)验证其与JEV的特异结合。结果 Co-IP复合物经SDS-PAGE分离出多个条带,选择最明显的3条带进行了质谱分析,结果显示分别为真核细胞翻译延长因子2(eEF2)、热休克蛋白90β(HSP90β)和微管蛋白α6。利用兔抗人HSP90β单克隆抗体进行Western blotting和ELISA实验,证实了细胞膜中确有HSP90β且可与JEV特异性结合。结论 HSP90β是HeLa细胞膜JEV受体的候选分子。Objective To identify proteins in HeLa cell membrane that can bind Japanese encephalitis virus （JEV）. Methods Co-immunoprecipitation between JEV and protein extracts of HeLa cell membrane was undertaken, and novel proteins capable of binding JEV suggested by mass spectrometric analysis were subject to Western blotting and ELISA. Results The most conspicuous proteins separated by Co-IP were eukaryotic elongation factor 2 （eEF2）, heat shock protein 90 kDa 1 beta （HSP9013）, and tubulin α6.Further Western blotting and ELISA by monoclonal antibodies verified that HSP9013 in HeLa cell membrane extracts bound JEV specifically. Conclusion HSP90β is a candidate receptor protein for JEV on HeLa cell membrane.

关 键 词：日本脑炎病毒 细胞膜蛋白 热休克蛋白90β 

分 类 号：R512.32[医药卫生—内科学]