乙型肝炎病毒核心抗原在人肝癌细胞株HepG2中的相互作用蛋白研究  

Screening of hepatitis B virus core antigen interactive protein in human HepG2 cells

作　　者：周剑[1] 刘东[2] 

机构地区：[1]解放军第三军医大学基础部,重庆400038 [2]解放军第三军医大学免疫学研究所,重庆400038

出　　处：《免疫学杂志》2014年第6期516-518,共3页Immunological Journal

摘　　要：目的利用串联亲和纯化和液相质谱技术研究HBcAg在人肝癌细胞株HepG2中的相互作用蛋白。方法构建HBcAg重组质粒pMSCV-HBc,采用串联亲和纯化技术富集HepG2细胞中HBcAg结合蛋白,电泳分离免疫沉淀复合物并从胶中切取HBcAg结合蛋白条带,胶内酶解后进行液相质谱技术分析从而鉴定HBcAg相互作用蛋白。结果成功构建HBcAg重组表达载体,在HepG2细胞中筛选到4个HBcAg相互用蛋白:CD59糖蛋白前体、MCM3相关蛋白、LOC100049716蛋白和ANKRD26蛋白。结论人肝癌细胞中HBcAg相互作用蛋白CD59等可能在HBV感染中起到重要作用。To investigate the interactive proteins of HBeAg in HepG2 cells, we constructed pMSCV-HBc expression vector and transformed the plasmids to HepG2 cells. Total proteins were enriched by tandem affinity purification and then separated on 10% SDS-polyaerylamide gel. The HBcAg binding proteins were cut from the gel and LC/MS methods were used to screen the HBcAg interactive proteins. The corresponding peptide sequence were obtained using database. Four interactive proteins were identified： CD59 glycoprotein precursor, MCM3-associated protein, LOC100049716 protein and ANKRD26-1ike family interactive proteins like CD59 may play important roles in HBV C member lB. The results showed that HBcAg infection pathogenesis.

关 键 词：乙肝核心抗原 相互作用蛋白 串联亲和纯化 液相质谱技术 

分 类 号：R735.7[医药卫生—肿瘤]