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

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 SAXS将用于了解乙肝病毒核心(由大肠杆菌表达的蛋白产生的非传染性病毒样颗粒)的组装和拆解，并研究抗病毒药物诱导的核心颗粒破坏的机制。乙肝病毒是一种主要的病原体，也是人类癌症最普遍的致病因素之一。虽然已经有了有效的疫苗，但增加能够对抗病毒的抗病毒治疗药物仍然是人们非常感兴趣的事情。这些研究的目标是从机械上了解复杂的病毒组件是如何构建的，以及衣壳靶向药物化合物如何使组装反应偏离路径。我们最近研究了衣壳的解体过程，它显示了由计算模型预测的明显的动力学轮廓。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的存在下以二聚体的形式保持几天或更长时间。在下一步，我们计划对通过快速停流混合导致盐浓度跃升的组装进行时间分辨研究。我们期望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对组装动力学的影响。

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.