Core B: Macromolecular and Cellular Structure Core

核心B：高分子和细胞结构核心

• 批准号: 10304091
• 负责人: DAVID A. AGARD
• 金额: $ 35.87万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-27 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10304091
• 关键词: 2019-nCoV; Affect; Affinity; Air; Binding; Biochemical; Biochemistry; Biological Assay; Cells; Cellular Structures; Cellular biology; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Electron Microscopy; Fixatives; Gene Mutation; Genetic; Goals; Homeostasis; In Vitro; Light; Lysosomes; Mammalian Cell; Metabolism; Methodology; Methods; Molecular; Molecular Chaperones; Molecular Conformation; Molecular Structure; Mutation; Organelles; Oxides; Peptide Hydrolases; Process; Proteins; Resolution; Sampling; Seeds; Structure; Technology; Therapeutic Intervention; Thinness; Tissues; Water; age related; cell behavior; extracellular; genetic variant; graphene; innovation; insight; interest; magnetic beads; novel; novel strategies; preservation; protein complex; protein expression; protein structure; reconstitution; structural biology; tau Proteins

PROJECT SUMMARY The primary objective of the Macromolecular and Cellular Structure Core is to provide cutting edge, innovative platforms for structural characterization responsive to needs and discoveries of the U54 FTD Center without Walls. The long-term goal of this FTD Center without Walls is to understand the overall metabolism of tau, how it is stabilized by chaperones and cochaperones and how it is channeled for degradation by the lysosome. This core will use the latest methodologies in cryoEM to determine atomic resolution cryoEM structures of relevant proteins and protein complexes as defined in Projects 1, 2 and to provide a cellular context for tau lysosomal degradation via cryoEM-Tomography. The high-resolution studies will be enabled via the expression and in vitro reconstitution of critical protein-protein complexes and novel cryoEM grid technologies developed by the Agard lab. Additionally, the core will use its biochemical expertise to quantify the existence of tau seeds for the other parts of the Center via RTQuiC assays. Together, the contributions from this Core will be significant as they will reveal fundamental mechanisms dictating tau turnover and provide novel targets for potential therapeutic intervention.

项目概要 高分子和细胞结构核心的主要目标是提供尖端、 响应 U54 FTD 中心的需求和发现的结构表征创新平台 没有墙。这个无墙 FTD 中心的长期目标是了解 tau，它如何被伴侣蛋白和共伴侣蛋白稳定，以及它如何被引导降解 溶酶体。该核心将使用冷冻电镜的最新方法来确定冷冻电镜的原子分辨率 项目 1、2 中定义的相关蛋白质和蛋白质复合物的结构，并提供细胞 通过冷冻电镜断层扫描了解 tau 蛋白溶酶体降解的背景。高分辨率研究将通过 关键蛋白质-蛋白质复合物和新型冷冻电镜网格的表达和体外重建 Agard 实验室开发的技术。此外，核心将利用其生化专业知识来量化 通过 RTQuiC 检测发现该中心其他部分存在 tau 种子。共同做出贡献 这个核心的成果将具有重要意义，因为它们将揭示决定 tau 更新的基本机制并提供 潜在治疗干预的新目标。