TIME-RESOLVED SOLUTION X-RAY SCATTERING STUDIES ON THE HEPATITIS B CAPSID PROTEI

乙型肝炎衣壳蛋白的时间分辨溶液X射线散射研究

• 批准号: 8362056
• 负责人: HIROTSUGU TSURUTA
• 金额: $ 0.14万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362056
• 关键词: Antiviral Agents; Capsid; Capsid Proteins; Complex; Computer Simulation; Drug Compounding; Escherichia coli; Exhibits; Funding; Future; Goals; Grant; Hepatitis B; Hepatitis B Virus; Human; Kinetics; Malignant Neoplasms; Measurement; National Center for Research Resources; Nucleosome Core Particle; Pathway interactions; Pharmaceutical Preparations; Principal Investigator; Process; Proteins; Proteus; Radiation; Reaction; Research; Research Infrastructure; Resources; Roentgen Rays; Shunt Device; Sodium Chloride; Solutions; Source; Therapeutic; Time; Time Study; United States National Institutes of Health; Urea; Vaccines; Viral; Virus; Virus-like particle; combat; cost; interest; pathogen; structural biology; virus core

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. SAXS will be applied to understand the assembly and disassembly of Hepatitis B virus (HBV) cores (non-infectious virus-like particles produced from E. coli expressed protein) and to investigate the mechanism of anti-viral drug-induced core particle disruption. HBV is a major pathogen and one of the most prevalent causative agents of cancer in humans. While effective vaccines are available, it remains of significant interest to add to the battery of antiviral therapeutics that can combat the virus. The goal of the studies is to understand mechanistically how a complex viral assembly is constructed and how capsid-targeted drug compounds can shunt the assembly reactions off-pathway. We recently studied the capsid disassembly process which exhibited a distinct kinetic profile predicted by computational models. We also conducted static x-ray scattering measurements of the capsid protein as a function of urea concentration prior to planned time-resolved measurements. We have been able to determine that the capsid protein can be kept unassembled in the dimeric form in the presence of 50 mM for a few days or longer. In the next step, we plan on conducing time-resolved studies of assembly induced a salt concentration jump via a rapid stopped-flow mixing. We anticipate to observe initial steps of the capsid assembly as a function of time and study the effects of potential drug compounds modifying assembly kinetics in near future.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 SAXS将用于了解乙型肝炎病毒（HBV）核心的组装和拆卸（非感染性病毒样颗粒，由大肠杆菌表达的蛋白质产生），并研究抗病毒药物诱导的核心颗粒粒子破坏的机制。 HBV是主要的病原体，也是人类癌症最普遍的病原体之一。尽管有有效的疫苗，但增加可以抵抗病毒的抗病毒疗法的电池仍然引起了人们的关注。研究的目的是从机械上理解如何构建复杂的病毒装配方式，以及靶向带状靶向的药物化合物如何将组装反应分流到沿途径的情况下。 我们最近研究了Capsid拆卸过程，该过程表现出了由计算模型预测的独特动力学曲线。我们还在计划的时间分辨测量之前，进行了衣壳蛋白的静态X射线散射测量，是尿素浓度的函数。我们已经能够确定可以在50 mm的情况下以二聚体形式将衣壳蛋白纯化几天或更长时间。在下一步中，我们计划通过快速停止流量混合促进组装的时间分辨研究诱导盐浓度跳跃。我们预计将观察带封圈组装的初始步骤是时间的函数，并研究不久的将来修饰组装动力学的潜在药物化合物的影响。