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

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心赠款提供。次级项目的主要支助 子项目的主要研究者可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 表示子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 SAXS将用于了解B型肝炎病毒（HBV）核心（由大肠杆菌产生的非感染性病毒样颗粒）的组装和拆卸。coli表达蛋白），并探讨抗病毒药物诱导核心颗粒破裂的机制。HBV是一种主要的病原体，也是人类癌症最普遍的致病因子之一。虽然有效的疫苗是可用的，但增加可以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械上了解复杂的病毒组装体是如何构建的，以及captain靶向药物化合物如何使组装反应偏离途径。 最近，我们研究了衣壳拆卸过程中表现出不同的动力学轮廓预测的计算模型。我们还进行了静态X射线散射测量的衣壳蛋白作为尿素浓度的函数之前，计划的时间分辨测量。我们已经能够确定衣壳蛋白可以在50 mM存在下以二聚体形式保持未组装数天或更长时间。在下一步中，我们计划进行时间分辨的研究，通过快速停流混合组装诱导盐浓度跳跃。我们期望观察作为时间函数的衣壳组装的初始步骤，并在不久的将来研究潜在药物化合物修饰组装动力学的影响。

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.