An integrated proteomics and bioinformatics analyses of hepatitis B virus X interacting proteins and identification of a novel interactor apoA-I
An integrated proteomics and bioinformatics analyses of hepatitis B virus X interacting proteins and identification of a novel interactor apoA-I
复制标题
乙型肝炎病毒 X 相互作用蛋白的综合蛋白质组学和生物信息学分析以及新型相互作用蛋白 apoA-I 的鉴定
DOI：
10.1016/j.jprot.2013.03.028
复制
发表时间：
2013-06-12
影响因子：
3.3
通讯作者：
Wei, Yuquan
中科院分区：
文献类型：
--
作者：
Zhang, Tao;Xie, Na;Wei, Yuquan
HBx是HBV编码的多功能蛋白，其生物学功能主要依赖于多效蛋白-蛋白相互作用（PPIs），但迄今为止尚未建立HBx相互作用组的整体图谱。因此，在本研究中，我们已经确定了127 HBx相互作用蛋白的深刻GST下拉结合质谱分析，并构建了HBx相互作用网络和核心apoA-I通路的一系列生物信息学方法。其中一种已鉴定的HBx结合伴侣是载脂蛋白A-I（apoA-I），其在脂质和胆固醇代谢中具有特定作用。通过GST pull-down和免疫共沉淀证实了HBx-apoA-I蛋白相互作用。apoA-I的异位过表达可导致HBV分泌的显著抑制，并伴随细胞胆固醇水平的降低。此外，HBV还可通过HBx调节apoA-I的功能，HBx可能与apoA-I的44-189位残基相互作用，导致apoA-I的功能障碍，如自结合能力降低、羰基水平升高和脂质结合能力受损。我们的研究结果证实了HBx与宿主蛋白质之间存在完整的物理结合，特别是一种新的相互作用因子apoA-I，它可能影响HBV的分泌，这为探讨HBV操纵宿主细胞功能的复杂机制提供了新的思路。虽然已经鉴定了一系列HBx相互作用蛋白，但尚未报道HBx相互作用组的整体表征。在这项研究中，我们已经确定了总共127 HBx相互作用蛋白的深刻GST下拉分析结合质谱，并构建了一个HBx相互作用网络与一系列的生物信息学方法。我们的研究结果表明，HBx和宿主蛋白质的整合物理协会，这可能有助于我们探索HBV操纵宿主细胞功能的复杂机制。此外，我们还验证了其中一种已鉴定的HBx结合伴侣载脂蛋白A-I（apoA-I），它对HBV分泌具有显著的抑制作用，表明HBx-apoA-I轴在HBV生命周期中起着至关重要的作用。(C)2013 Elsevier B. V.保留所有权利。
HBx is well-known to be a multifunctional protein encoded by HBV and its biological functions are mainly dependent on pleiotropic protein-protein interactions (PPIs); however, the global mapping of HBx-interactome has not been established so far. Thus, in this study, we have identified 127 HBx-interacting proteins by a profound GST pull-down assay coupled with mass spectrometry, and constructed an HBx-interactome network and core apoA-I pathways with a series of bioinformatics approaches. One of the identified HBx-binding partners is apolipoprotein A-I (apoA-I), which has a specific role in lipid and cholesterol metabolism. The HBx-apoA-I protein interaction was confirmed by both GST pull-down and co-immunoprecipitation. The ectopic overexpression of apoA-I can lead to a significant inhibition on HBV secretion concomitant with the reduction of cellular cholesterol level. In addition, HBV can modulate the function of apoA-I through HBx which might interact with the 44-189 residues of apoA-I and result in dysfunction of apoA-I such as decreased self-association ability, increased carbonyl level and impaired lipid-binding ability. Our results demonstrate an integrated physical association of HBx and host proteins, especially a novel interactor apoA-I that may influence the HBV secretion, which would shed new light on exploring the complicated mechanisms of HBV manipulation on host cellular functions.Biological significanceHBx is well-known to be a multifunctional protein encoded by HBV and its biological functions are mainly dependent on pleiotropic protein-protein interactions. Although a series of HBx-interacting proteins have been identified, a global characterization of HBx interactome has not been reported. In this study, we have identified a total of 127 HBx-interacting proteins by a profound GST pull-down assay coupled with mass spectrometry, and constructed an HBx-interactome network with a series of bioinformatics approaches. Our results demonstrate an integrated physical association of HBx and host proteins which may help us explore the complicated mechanisms of HBV manipulation on host cellular functions. In addition, we validated one of the identified HBx-binding partners, apolipoprotein A-I (apoA-I), which played a significant inhibitory effect on HBV secretion, indicating a crucial role of the HBx-apoA-I axis in HBV life cycle. (C) 2013 Elsevier B.V. All rights reserved.