Core 1 - Structural Biology Core

核心 1 - 结构生物学核心

• 批准号: 10506664
• 负责人: Priyamvada Acharya
• 金额: $ 89.03万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-14 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10506664
• 关键词: AIDS/HIV problem; B-Lymphocytes; Cells; Communities; Cryo-electron tomography; Cryoelectron Microscopy; Drug Design; Event; Evolution; Future; Goals; HIV; HIV Envelope Protein gp120; HIV-1; Heterogeneity; Image Analysis; Immune system; Infection; Infrastructure; Kinetics; Light; Maps; Mediating; Methods; Mission; Modeling; Monitor; Persons; Preparation; Process; Protocols documentation; Research; Research Project Grants; Resolution; Roentgen Rays; Sampling; Series; Solid; Sorting - Cell Movement; Specimen; Structure; T-Cell Activation; Techniques; Technology; Temperature; Time; Tomogram; Vaccines; Viral; Virus; Virus Diseases; antiretroviral therapy; biological systems; computerized tools; cryogenics; env Gene Products; expectation; improved; in situ imaging; millisecond; movie; particle; structural biology; technology development; temperature jump; temporal measurement; tool; viral rebound

Abstract – Core 1 – Structural Biology Core Approximately 40 million people worldwide are living with HIV/AIDS; however, a protective vaccine or functional cure remain elusive despite four decades of intense research. HIV-1 evades the immune system through its rapid structural evolution during infection and replication. The Duke Center for HIV Structural Biology will pursue structural studies of the evolution of the HIV-1 Envelope (Env) protein to elucidate structure-function mechanisms for viral entry, B-cell and T-cell activation, and viral rebound after antiretroviral therapy ART. The Structural Biology Core (Core 1) will support the overall mission of the Center by establishing a state-of-the-art pipeline for structural analysis of HIV-1 Env using a wide range of experimental techniques. The Core will provide access to cutting-edge techniques for structure determination and have a strong component of technology development that will ultimately advance our mechanistic understanding of Env. The research projects will benefit from having access to established protocols for structure determination as well as new methods resulting from the technology development efforts of the core. The Specific Aims of the Structural Biology Core are 1) to establish a high- throughput pipeline for routine characterization of the structure and dynamics of soluble HIV-1 trimers using high- resolution single-particle cryo-EM; 2) to develop advanced workflows for structural analysis of native HIV-1 samples imaged in-situ using cryo-electron tomography (ET) at near-atomic resolution; and 3) to establish structural methods for microsecond time resolution structural studies of HIV-1 Env. Completion of the three proposed aims will provide a solid infrastructure in structural biology needed to support the overall goals of the Center and its components. By providing access to state-of-the-art technology for the determination of structures of HIV-1 at the highest possible spatial and temporal resolution, will provide unique opportunities for visualizing key intermediates that could inform our understanding of HIV viral infection. The ability to determine high-resolution structures of soluble or native forms of Env will be critical to improve our understanding of HIV. The technologies developed as part of the Structural Biology Core will have implications beyond the field of HIV and would benefit structural studies of other biological systems. By making our tools available to the structural biology community, the activities of the core will have an even wider impact.

核心1 -结构生物学核心 全世界约有4000万人感染艾滋病毒/艾滋病;然而，保护性疫苗或功能性疫苗 尽管进行了40年的深入研究，但治愈方法仍然难以捉摸。HIV-1通过其 感染和复制期间的快速结构进化。杜克艾滋病毒结构生物学中心将继续研究 HIV-1包膜蛋白进化的结构研究，以阐明结构-功能机制 用于病毒进入、B细胞和T细胞活化以及抗逆转录病毒治疗ART后的病毒反弹。 生物核心（核心1）将通过建立一个最先进的管道， 使用广泛的实验技术进行HIV-1 Env的结构分析。核心将提供访问 尖端技术的结构确定和技术开发的强大组成部分 这将最终推进我们对Env的机械理解。研究项目将受益于 获得已建立的结构测定方案以及该技术产生的新方法 核心的发展努力。结构生物学核心的具体目标是1）建立一个高- 高通量流水线，用于可溶性HIV-1三聚体的结构和动力学的常规表征， 分辨率单粒子冷冻EM; 2）开发用于天然HIV-1结构分析的先进工作流程 使用近原子分辨率的低温电子断层扫描（ET）对样品进行原位成像;以及3）建立 HIV-1包膜蛋白微秒级时间分辨结构研究方法完成三个 拟议的目标将提供一个坚实的基础设施，在结构生物学需要支持的总体目标， 中心及其组件。通过提供最先进的技术来确定结构 在尽可能高的空间和时间分辨率下对HIV-1的研究，将为可视化提供独特的机会， 关键的中间体，可以让我们了解艾滋病毒感染。 确定可溶性或天然形式的Env的高分辨率结构的能力对于改善我们的研究至关重要。 了解艾滋病。作为结构生物学核心的一部分开发的技术将产生影响 这将超越艾滋病毒领域，并将有利于其他生物系统的结构研究。通过制造工具 由于结构生物学界可利用的资源有限，核心小组的活动将产生更广泛的影响。