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: 8169937
• 负责人: HIROTSUGU TSURUTA
• 金额: $ 0.24万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169937
• 关键词: Antiviral Agents; Capsid; Capsid Proteins; Complex; Computer Retrieval of Information on Scientific Projects Database; Computer Simulation; Drug Compounding; Escherichia coli; Exhibits; Funding; Future; Goals; Grant; Hepatitis; Hepatitis B Virus; Human; Institution; Kinetics; Malignant Neoplasms; Measurement; Nucleosome Core Particle; Pathway interactions; Pharmaceutical Preparations; Process; Proteins; Proteus; Reaction; Research; Research Personnel; Resources; Shunt Device; Sodium Chloride; Solutions; Source; Therapeutic; Time; Time Study; United States National Institutes of Health; Urea; Vaccines; Viral; Virus; Virus-like particle; combat; interest; pathogen; virus core

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 SAXS 将用于了解乙型肝炎病毒 (HBV) 核心（由大肠杆菌表达的蛋白产生的非传染性病毒样颗粒）的组装和分解，并研究抗病毒药物诱导核心颗粒破坏的机制。乙型肝炎病毒是一种主要病原体，也是人类最常见的癌症病原体之一。尽管已经有了有效的疫苗，但添加可对抗病毒的抗病毒疗法仍然具有重大意义。这些研究的目的是从机制上了解复杂的病毒组装体是如何构建的，以及衣壳靶向药物化合物如何将组装反应分流。 我们最近研究了衣壳分解过程，该过程表现出计算模型预测的独特动力学特征。在计划的时间分辨测量之前，我们还对衣壳蛋白进行了静态 X 射线散射测量，作为尿素浓度的函数。我们已经能够确定衣壳蛋白可以在 50 mM 存在下保持未组装的二聚体形式几天或更长时间。下一步，我们计划对通过快速停流混合引起的盐浓度跳跃进行装配的时间分辨研究。我们预计在不久的将来观察衣壳组装的初始步骤作为时间的函数，并研究潜在的药物化合物改变组装动力学的影响。