北京蛋白質(zhì)組研究中心/蛋白質(zhì)組學(xué)國家重點實驗室姜穎副研究員課題組劉坤博士等發(fā)現(xiàn)乙型肝炎病毒相關(guān)疾病治療的潛在靶點,，相關(guān)論文發(fā)表在最新一期國際蛋白質(zhì)組學(xué)權(quán)威雜志：《分子與細胞蛋白質(zhì)組學(xué)》（Molecular & Cellular Proteomics, MCP）上面，同期雜志還發(fā)表了該所朱云平研究員課題組、錢小紅研究員課題組的兩篇研究論文,，創(chuàng)該刊單期同一單位發(fā)文數(shù)之最。
乙型肝炎病毒（HBV）感染是一種嚴(yán)重危害人類健康的重大疾病，目前治療手段有限，其重要原因是缺乏有效的治療靶點,。他們用先進的蛋白質(zhì)復(fù)合體分離和鑒定方法，發(fā)現(xiàn)熱休克蛋白HSP90 和HSP70/HSP60形成的內(nèi)源性分子伴侶復(fù)合體參與了HBV的復(fù)制及分泌,，并用RNAi和小分子抑制劑證明它們可作為治療HBV相關(guān)疾病的潛在靶點,。此研究結(jié)果為系統(tǒng)了解HBV的生命周期及研發(fā)相關(guān)疾病的治療藥物提供了新的思路。（生物谷Bioon.com）
生物谷推薦原始出處：
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.
