北京蛋白质组研究中心/蛋白质组学国家重点实验室姜颖副研究员课题组刘坤博士等发现乙型肝炎病毒相关疾病治疗的潜在靶点，相关论文发表在最新一期国际蛋白质组学权威杂志：《分子与细胞蛋白质组学》（Molecular & Cellular Proteomics, MCP）上面，同期杂志还发表了该所朱云平研究员课题组、钱小红研究员课题组的两篇研究论文，创该刊单期同一单位发文数之最。
乙型肝炎病毒（HBV）感染是一种严重危害人类健康的重大疾病，目前治疗手段有限，其重要原因是缺乏有效的治疗靶点。他们用先进的蛋白质复合体分离和鉴定方法，发现热休克蛋白HSP90 和HSP70/HSP60形成的内源性分子伴侣复合体参与了HBV的复制及分泌，并用RNAi和小分子抑制剂证明它们可作为治疗HBV相关疾病的潜在靶点。此研究结果为系统了解HBV的生命周期及研发相关疾病的治疗药物提供了新的思路。（生物谷Bioon.com）
生物谷推荐原始出处：
Molecular & Cellular Proteomics 8:495-505, 2009.doi:10.1074/mcp.M800250-MCP200
Two-dimensional Blue Native/SDS-PAGE Analysis Reveals Heat Shock Protein Chaperone Machinery Involved in Hepatitis B Virus Production in HepG2.2.15 Cells
Kun Liu,, Lu Qian?, Jinglan Wang, Wenrui Li, Xinyu Deng, Xilin Chen, Wei Sun, Handong Wei, Xiaohong Qian, Ying Jiang,|| and Fuchu He,**,
From the State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China, ** Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China, and ? Department of Cellular Immunology, Institute of Basic Medical Science, Beijing 100850, China
Hepatitis B virus (HBV) infection is a major health concern with more than two billion individuals currently infected worldwide. Despite the prevalence of infection, gaining a complete understanding of the molecular mechanisms of HBV infection has been difficult because HBV cannot infect common immortalized cell lines. HepG2.2.15, however, is a well established version of the HepG2 cell line that constitutively expresses HBV. Therefore, comparative proteomics analysis of HepG2.2.15 and HepG2 may provide valuable clues for understanding the HBV virus life cycle. In this study, two-dimensional blue native/SDS-PAGE was utilized to characterize different multiprotein complexes from whole cell lysates between HepG2.2.15 and HepG2. These results demonstrate that two unique protein complexes existed in HepG2.2.15 cells. When these complexes were excised from the gel and subjected to the second dimension separation and the proteins were sequenced by mass spectrometry, 20 non-redundant proteins were identified. Of these proteins, almost 20% corresponded to heat shock proteins, including HSP60, HSP70, and HSP90. Antibody-based supershift assays were used to verify the validity of the distinct protein complexes. Co-immunoprecipitation assays confirmed that HSP60, HSP70, and HSP90 proteins physically interacted in HepG2.2.15 but not HepG2 cells. We further demonstrated that down-regulation of HSP70 or HSP90 by small interfering RNA significantly inhibited HBV viral production but did not influence cellular proliferation or apoptosis. Consistent with these results, a significant reduction in HepG2.2.15 HBV secretion was observed when the HSP90 inhibitor 17-allylamino-17-demethoxygeldanamycin was used to treat HepG2.2.15 cells. Collectively these results suggest that the interaction of HSP90 with HSP70/HSP60 contributes to the HBV life cycle by forming a multichaperone machine that may constitute therapeutic targets for HBV-associated diseases.
