CHEETAH Center for the Structural Biology of HIV Infection, Restriction, and Viral Dynamics

CHEETAH HIV 感染、限制和病毒动力学结构生物学中心

• 批准号: 10663355
• 负责人: JOSEPH D PUGLISI
• 金额: $ 32.73万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-11 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10663355
• 关键词: Antibodies; Behavior; Binding; Biological; Biological Process; Biology of HIV Infection; Complex; Cryoelectron Microscopy; Electron Microscopy; Electron energy loss spectroscopy; Electrons; Environment; Epitopes; Fluorescence; Fluorescence Spectroscopy; HIV; HIV-1; Hybrids; Image; Investigation; Ions; Label; Link; Macromolecular Complexes; Measures; Methodology; Methods; Microscopy; Molecular; Molecular Conformation; Nanotechnology; Proteins; RNA; RNA I; Reporter; Research Personnel; Resolution; Site; Structure; System; Technology; Time; Viral; Virus; Visualization; antibody mimetics; biological systems; design; empowerment; imaging approach; instrumentation; light microscopy; member; novel; novel strategies; particle; protein complex; scaffold; single molecule; spectroscopic imaging; structural biology; three dimensional structure; tool; trafficking; waveguide

CORE SUMMARY This Core is designed to provide CHEETAH Center members with access to novel approaches for imaging and measuring the behaviors of different HIV-host systems under investigation in our Center. Specifically, our Tools for Structural and Mechanistic Studies Core (Core 2), will develop new methodology and provide CHEETAH Center members with access to state-of-the-art instrumentation and expertise in Fluorescence Spectroscopy and Dynamics, Scaffolds for Structure Determination, and Electron Cryotomography. Component 1 (Fluorescence Spectroscopy and Dynamics) will provide instrumentation and expertise in fluorescence labeling and single-molecule fluorescence approaches to studies of biological structure and dynamics. Novel methods for correlating biological processes in real time and for linking cryoEM structural studies to dynamic analyses are also being developed. Component 2 (Scaffolds for Structure Determination) will generate and provide antibody-like molecules called “Synthetic Antigen Binders” (sABs) as fiducial markers for cryoEM structure determinations. The Component will also optimize and apply new, empowering classes of “universal” molecular fiducials based on: 1) transportable binding epitopes, or 2) secondary antibodies. Universal fiducials for more challenging RNAs and RNA-protein complexes are also being created. Component 3 (Electron Cryotomography) will incorporate the powerful and rapidly advancing technologies of correlative light and electron microscopy (CLEM), focused ion beam (FIB) milling, and electron cryotomography (ECT) to reconstruct native molecular landscapes at unprecedented resolution. The Component will also develop new functional correlative reporters that can be located by fluorescence or by electron energy-loss spectroscopy (EELS) imaging, and apply these reporters to problems in HIV-1 sub-viral organization, virus entry, and viral core replication and trafficking.

核心摘要 该核心旨在为 CHEETAH 中心成员提供新的成像和测量方法 我们中心正在研究不同艾滋病毒宿主系统的行为，特别是我们的结构和工具。 机制研究核心（核心 2），将开发新的方法并为 CHEETAH 中心成员提供访问 荧光光谱和动力学、结构支架方面最先进的仪器和专业知识 测定和电子冷冻断层扫描。 第 1 部分（荧光光谱和动力学）将提供荧光方面的仪器和专业知识 用于研究生物结构和动力学的新方法的标记和单分子荧光方法。 实时关联生物过程以及将冷冻电镜结构研究与动态分析联系起来也正在研究中。 发达。 组件 2（用于结构测定的支架）将生成并提供称为“合成抗体”的抗体样分子。 抗原结合剂”（sAB）作为冷冻电镜结构测定的基准标记，该组件也将得到优化和优化。 应用新的、授权的“通用”分子基准类别，基于：1）可运输的结合表位，或2） 用于更具挑战性的 RNA 和 RNA-蛋白质复合物的通用基准也正在创建中。 第 3 部分（电子冷冻断层扫描）将结合强大且快速发展的相关技术 光学和电子显微镜 (CLEM)、聚焦离子束 (FIB) 铣削和电子冷冻断层扫描 (ECT) 来重建 该组件还将开发新的功能相关物。 可以通过荧光或电子能量损失光谱 (EELS) 成像定位的生产者，并应用这些 在 HIV-1 亚病毒组织、病毒进入以及病毒核心复制和贩运方面产生问题。