Project 3: Structural basis of amyloid formation and chaperone-mediated turnover

项目 3：淀粉样蛋白形成和分子伴侣介导的周转的结构基础

基本信息

批准号：
10377430
负责人：
Daniel Ryland Southworth
金额：
$ 35.53万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10377430
关键词：
Address Adopted Affinity Aging Alzheimer&apos s Disease Amyloid Amyloid Fibrils Amyloid beta-Protein Animal Model Animals Antibodies Binding Biological Models Brain Cell Culture Techniques Cells Characteristics Complex Cryoelectron Microscopy Data Dementia Disease Disease Progression Disease model Filament Film Future Goals Heat-Shock Proteins 70 Human In Vitro K-18 conjugate Length Mediating Methods Modeling Modification Molecular Molecular Chaperones Molecular Conformation Molecular Structure Mus Mutation Neurodegenerative Disorders Neurofibrillary Tangles Nucleotides Oxides Pathologic Peptides Pharmacology Population Prions Process Protein Isoforms Proteins Quality Control Regulation Resolution Senile Plaques Source Structure Surface System Technology Therapeutic Tissues Toxic effect Ubiquitination Variant Work advanced disease amyloid formation amyloid structure base beta pleated sheet brain tissue chaperone machinery crosslink disease-causing mutation experience functional group genetic regulatory protein graphene human tissue in vivo interest mouse model mutant new therapeutic target novel therapeutic intervention prevent response small molecule tau Proteins tau aggregation ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY Currently there are no known treatments that slow or prevent Alzheimer’s and other neurodegenerative diseases, and the molecular mechanisms that underlie the progression of these diseases are poorly understood. Our broad goal is to address critical problems in macromolecular structure determination of disease-relevant conformations of Aβ and tau prions and associated protein regulatory complexes in order to advance mechanistic understanding of prion propagation and guide novel therapeutic approaches. We will achieve this goal using high-resolution cryo-electron microscopy (cryo-EM) methods and: (1) Develop affinity capture methods on cryo-EM grids for extracting specific filaments from tissue for structure determination; (2) Determine cryo-EM structures of Aβ and tau fibril conformations derived from in vitro assembly and mouse models and compare with those determined from human tissue; and (3) Determine mechanisms and interactions by the Hsp70 molecular chaperone machinery that regulate prion propagation, ubiquitination, and clearance. With these goals we will identify the structural basis for amyloid fibrils that develop during disease and uncover key chaperone regulatory processes critical for quality control and clearance of proteins that form toxic amyloids.

项目概要目前尚无已知的治疗方法可以减缓或预防阿尔茨海默病和其他神经退行性疾病疾病，以及这些疾病进展的分子机制尚不清楚我们的总体目标是解决大分子结构测定中的关键问题。 Aβ 和 tau 朊病毒以及相关蛋白质调节复合物的疾病相关构象，以便我们将推进对朊病毒传播机制的理解并指导新的治疗方法。使用高分辨率冷冻电子显微镜 (cryo-EM) 方法实现这一目标，并且：(1) 开发亲和力 (2) 冷冻电镜网格上的捕获方法，用于从组织中提取特定丝以进行结构测定；确定来自体外组装和小鼠的 Aβ 和 tau 原纤维构象的冷冻电镜结构模型并与从人体组织中确定的结果进行比较；以及 (3) 确定机制和 Hsp70 分子伴侣机制的相互作用，调节朊病毒的传播、泛素化和通过这些目标，我们将确定疾病期间形成的淀粉样原纤维的结构基础。并揭示对形成的蛋白质的质量控制和清除至关重要的关键伴侣监管过程有毒的淀粉样蛋白。