STRUCTURAL BASIS OF CONTROLLING VIRUS CAPSID ASSEMBLY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 了解病毒衣壳的组装对于理解病毒生物学、开发组装导向的抗病毒疗法和发展纳米技术具有重要的应用价值。在这项提案中，我们要求有时间收集有关乙肝病毒衣壳与影响组装的小分子复合的数据。这将定义这些组装效应器的各自结合部位，并将允许我们推导出一种作用机制。我们还将获得组装在纳米颗粒核心上的植物病毒衣壳的结构数据。这种结构将提供对病毒-核心相互作用的洞察，同样重要的是，为确定这种杂化颗粒结构的实验特性提供了一个试验台。 超过3.5亿人患有慢性乙肝，其中100万人在美国。每年有60万人死于乙肝病毒。我们已经确定了乙肝病毒衣壳的组装是抗病毒的靶点。在体内，正确的组装对病毒复制至关重要。最近，我们用小分子组装效应器确定了乙肝病毒衣壳的结构，并能够确定结合位置并阐明了活性的热力学基础。我们已经确定了一种新的效应器家族，可以诱导比正常衣壳更小的衣壳组装。这些复合体的晶体在原始源上的衍射率高达5.5？；通常，我们使用同步辐射将病毒晶体的分辨率提高了1.5到2.5？ 我们还研究了基于病毒的纳米结构的组装。在与Bogdan Dragnea(也是在印第安纳大学)的合作下，我们获得了在体外组装在聚乙二醇化的金纳米颗粒上的雀麦花叶病毒衣壳蛋白的晶体。在这里，我们可以问蛋白质和底物的相互作用有多密切。 这些实验中使用的所有样本都是类似病毒的颗粒，而不是传染性病毒粒子。

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. Understanding assembly of virus capsids has applications for understanding virus biology, development of assembly-directed antiviral therapeutics, and development of nanotechnology. In this proposal we request time to collect data on hepatitis B virus (HBV) capsids complexed with small molecules that affect assembly. This will define the respective binding sites of these assembly effectors and will allow us to deduce a mechanism of action. We will also acquire data for the structure of a plant virus capsid that was assembled on a nanoparticle core. This structure will provide insight into the virus-core interaction and as importantly provide a testbed for defining the experimental peculiarities of determining the structure of such hybrid particles. More than 350 million individuals suffer from chronic HBV, 1 million in the United States. HBV contributes 600,000 deaths each year. We have identified assembly of the HBV capsid as an antiviral target. In vivo, correct assembly is critical to viral replication. Recently we determined the structure of HBV capsids with a small molecule assembly effector and were able to determine the binding site and elucidate the thermodynamic basis of activity. We have identified a new family of effectors that induce assembly of smaller than normal capsids. Crystals of these complexes have diffracted up to 5.5¿ resolution on a home source; typically we have improved resolution of virus crystals by 1.5 to 2.5¿ using synchrotron radiation. We have also investigated assembly of virus-based nanostructures. In collaboration with Bogdan Dragnea (also at Indiana University), we have obtained crystals of Brome Mosaic Virus capsid protein that were assembled in vitro on a PEGylated gold nanoparticle. Here we can ask how intimately the protein and substrate interact. All samples used in these experiments are virus-like particles, not infectious virions.