CONFORMATIONAL DYNAMICS OF ICOSAHEDRAL VIRUSES PROBED BY SAXS

通过 SAXS 探测二十面体病毒的构象动力学

• 批准号: 8169936
• 负责人: John Emil Johnson
• 金额: $ 0.34万
• 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/8169936
• 关键词: Address; Antiviral Agents; Bacteriophages; Capsid; Capsid Proteins; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA; Data; Equilibrium; Funding; Future; Goals; Grant; Head; Insect Viruses; Institution; Kinetics; Measures; Monitor; Nucleic Acids; Protein Subunits; RNA; Research; Research Personnel; Resolution; Resources; Signal Transduction; Solutions; Source; Time; United States National Institutes of Health; Virion; Virus; neutralizing antibody; receptor binding; reconstruction

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. The protein capsids of many viruses respond dynamically to environmental signals such as DNA or RNA packaging, receptor binding, and changes in pH or ionic composition. As a result, they undergo conformational changes that involve the concerted reorganization of hundreds of protein subunits and nucleic acid. Small-angle x-ray scattering (SAXS) is ideally suited to monitoring these structural changes in solution. We propose to use SAXS to monitor the dynamic conformational changes observed in two types of icosahedral virus particles, 1) an empty phage head capsid composed of 420 identical subunits (HK97 bacteriophage) and 2) a ssRNA insect virus composed of 240 identical capsid proteins and encapsidated RNA (N-omega-V tetravirus). The data we obtain will be used for generating low-resolution 3D reconstructions as well as for quantitative analysis of time-resolved structural changes using Singular Value Decomposition (SVD). The primary intellectual goals of the proposed studies are to measure the extent of cooperativity among subunits during large-scale conformational changes of these massive and complex assemblies, and to probe for the existence of kinetic and equilibrium intermediate states. By quantitatively addressing these issues we hope to better understand virus function and to enlighten future strategies for targeting capsids with antiviral compounds and neutralizing antibodies.

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。 许多病毒的蛋白质衣壳对环境信号做出动态响应，例如 DNA 或 RNA 包装、受体结合以及 pH 或离子组成的变化。 结果，它们经历构象变化，涉及数百个蛋白质亚基和核酸的协同重组。 小角 X 射线散射 (SAXS) 非常适合监测溶液中的这些结构变化。 我们建议使用SAXS来监测两种二十面体病毒颗粒中观察到的动态构象变化，1）由420个相同亚基（HK97噬菌体）组成的空噬菌体头衣壳和2）由240个相同衣壳蛋白组成的ssRNA昆虫病毒和衣壳化 RNA（N-omega-V 四病毒）。 我们获得的数据将用于生成低分辨率 3D 重建以及使用奇异值分解 (SVD) 对时间分辨结构变化进行定量分析。 拟议研究的主要智力目标是测量这些大规模复杂组装体的大规模构象变化期间亚基之间的协同程度，并探讨动力学和平衡中间态的存在。 通过定量解决这些问题，我们希望更好地了解病毒功能，并启发未来用抗病毒化合物和中和抗体靶向衣壳的策略。