Hepatitis C virus E2 glycoprotein

描述（由申请人提供）：丙型肝炎病毒（HCV）慢性感染全球2-3%的人口，使患者易患慢性肝病和肝癌。HCV感染最有效的治疗方法是使用聚乙二醇化干扰素- α (IFN-1)和利巴韦林的联合治疗，但这种治疗仅部分有效，费用昂贵，需要长期的医疗支持和随访。为了应对这一重大公共卫生威胁，对病毒分子的详细了解将极大地促进疫苗和抗病毒药物的设计。HCV包膜刺突由病毒糖蛋白E1和E2的异源二聚体组成，对于病毒附着和进入宿主细胞以及感染性病毒颗粒的组装非常重要。E1和E2糖蛋白是药理和免疫干预的潜在靶点。本应用的长期目标是确定HCV E2糖蛋白的晶体结构，以帮助合理设计抗病毒药物和疫苗，并在结构水平上了解病毒的进入机制。具体目标是：(i)开发生产高纯度重组E2蛋白的方法，并详细表征其生化特性；（ii）生成重组E2蛋白晶体，作为HCV E2糖蛋白晶体学研究的概念验证。这些目标的成功将为研究E2的高分辨率晶体结构提供坚实的科学基础。公共卫生相关性：测定丙型肝炎病毒（HCV） E2糖蛋白的晶体结构将为设计抗丙型肝炎病毒的药物和疫苗提供极有价值的信息。主要的科学障碍是缺乏高纯度和天然折叠的E2糖蛋白，这些糖蛋白将形成x射线衍射的晶体。本提案的目的是通过开发生产天然折叠E2的方法，并通过确定其结晶的实验条件，作为确定HCV E2糖蛋白高分辨率晶体结构的概念验证研究，来消除这一障碍。

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) chronically infects 2-3% of the global population, predisposing the patients to chronic liver diseases and liver cancer. The most effective treatment for HCV infection employs a combination of pegylated interferon-alpha (IFN-1) and ribavirin which is only partially effective, costly and requires long-term medical support and follow-up. To combat this major public health threat, a detailed understanding of the viral molecules will greatly facilitate vaccine and anti-viral drug design. The HCV envelope spikes, comprised of heterodimers of the E1 and E2 viral glycoproteins, are important for viral attachment and entry into host cells, and the assembly of infectious virus particles. The E1 and E2 glycoproteins are potential targets for pharmacological and immunological intervention. The long-term objective of this application is to determine the crystal structure of HCV E2 glycoprotein to aid the rational design of anti-viral drugs and vaccines, and to understand the viral entry mechanism at the structural level. The specific aims are (i) to develop methods to produce highly pure recombinant E2 proteins and to characterize their biochemical properties in details; and (ii) to generate a crystal of the recombinant E2 protein as a proof-of-concept for the crystallographic study of HCV E2 glycoprotein. Success in these aims will provide a strong scientific foundation for pursuing the high-resolution crystal structure of E2. PUBLIC HEALTH RELEVANCE: Determination of the crystal structure of the E2 glycoprotein of Hepatitis C virus (HCV) will provide extremely valuable information for drug and vaccine design to combat HCV. The major scientific roadblock is a lack of highly pure and natively folded E2 glycoproteins that will form crystals for X-ray diffraction. The aim of this proposal is to remove this roadblock by developing methods to produce natively folded E2 and by identifying the experimental conditions for its crystallization as a proof-of-concept study for the determination of high-resolution crystal structure of HCV E2 glycoprotein.