STRUCTURAL BASIS OF CONTROLLING VIRUS CAPSID ASSEMBLY

控制病毒衣壳组装的结构基础

• 批准号: 8171995
• 负责人: Adam Zlotnick
• 金额: $ 1.09万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171995
• 关键词: Affect; Antiviral Agents; Binding Sites; Biology; Brome mosaic virus; Capsid; Capsid Proteins; Cessation of life; Chronic Hepatitis B; Collaborations; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Development; Family; Funding; Gold; Grant; Hepatitis B Virus; Home environment; Hybrids; In Vitro; Indiana; Individual; Institution; Nanostructures; Nanotechnology; Plant Viruses; Proteins; Request for Proposals; Research; Research Personnel; Resolution; Resources; Sampling; Source; Structure; Thermodynamics; Time; United States; United States National Institutes of Health; Universities; Viral; Virion; Virus; Virus Assembly; Virus-like particle; base; improved; in vivo; insight; nanoparticle; particle; research study; small molecule; synchrotron radiation; therapeutic development; 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. 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.

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。子项目和 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。 了解病毒衣壳的组装可应用于了解病毒生物学、开发组装导向的抗病毒疗法以及开发纳米技术。 在此提案中，我们请求时间收集与影响组装的小分子复合的乙型肝炎病毒 (HBV) 衣壳的数据。 这将定义这些装配效应器各自的结合位点，并使我们能够推断出作用机制。 我们还将获取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深入了解，并且重要的是提供一个测试平台，用于定义确定此类混合颗粒结构的实验特性。 超过 3.5 亿人患有慢性乙型肝炎，其中美国有 100 万人。乙型肝炎每年导致 60 万人死亡。 我们已将 HBV 衣壳的组装确定为抗病毒靶点。在体内，正确的组装对于病毒复制至关重要。 最近，我们用小分子组装效应子确定了 HBV 衣壳的结构，并能够确定结合位点并阐明活性的热力学基础。我们已经确定了一个新的效应子家族，可以诱导比正常衣壳更小的组装。 这些配合物的晶体在家庭光源上的衍射分辨率高达 5.5°；通常，我们使用同步加速器辐射将病毒晶体的分辨率提高了 1.5 至 2.5°。 我们还研究了基于病毒的纳米结构的组装。 我们与 Bogdan Dragnea（同样在印第安纳大学）合作，获得了雀麦花叶病毒衣壳蛋白晶体，该晶体在体外组装在聚乙二醇化金纳米颗粒上。 在这里我们可以问蛋白质和底物相互作用的密切程度。 这些实验中使用的所有样本都是病毒样颗粒，而不是传染性病毒颗粒。