Hepatitis C virus E2 glycoprotein

描述(申请人提供)：丙型肝炎病毒(丙型肝炎病毒)慢性感染全球人口的2-3%，使患者易患慢性肝病和肝癌。对丙型肝炎病毒感染最有效的治疗方法是聚乙二醇化干扰素-1和利巴韦林的组合，这种疗法只有部分有效，费用昂贵，需要长期的医疗支持和随访。为了对抗这一重大的公共卫生威胁，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。丙型肝炎病毒包膜尖峰由病毒糖蛋白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传染性病毒颗粒的组装是重要的。E1和E2糖蛋白是药物和免疫干预的潜在靶点。这一应用的长期目标是确定丙型肝炎病毒E2糖蛋白的晶体结构，以帮助抗病毒药物和疫苗的合理设计，并从结构水平上了解病毒的侵入机制。其具体目的是(I)建立制备高纯度重组E2蛋白的方法，并对其生化性质进行详细的鉴定；(Ii)制备重组E2蛋白的晶体，作为对HCVE2糖蛋白进行结晶学研究的概念验证。这些目标的成功将为追求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：丙型肝炎病毒(丙型肝炎病毒)E2糖蛋白晶体结构的测定将为抗丙型肝炎病毒的药物和疫苗设计提供极其有价值的信息。主要的科学障碍是缺乏高纯度和天然折叠的E2糖蛋白，这些糖蛋白将形成晶体用于X射线衍射。这项建议的目的是通过开发产生天然折叠的E2的方法并确定其结晶的实验条件来消除这一障碍，作为确定HCVE2糖蛋白高分辨率晶体结构的验证性研究。

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.