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.

项目摘要目前，没有已知的治疗方法可以缓慢或预防阿尔茨海默氏症和其他神经退行性疗法疾病以及这些疾病进展的基础的分子机制很差理解。我们的广泛目标是解决大分子结构确定的关键问题 Aβ和TAU prions及相关蛋白质调节复合物的疾病相关构象提前对prion传播的机理理解，并指导新颖的治疗方法。我们将使用高分辨率的低分子 - 电子显微镜（Cryo-EM）方法实现此目标，并且：（1）发展亲和力捕获用于从组织中提取特定细丝以进行结构测定的冷冻EM网格的方法；（2）确定源自体外组装和小鼠的Aβ和Tau原纤维组成的冷冻EM结构模型并与由人体组织确定的模型进行比较；（3）确定机制和 HSP70分子伴侣机械的相互作用，可调节prion传播，泛素化和清除。有了这些目标，我们将确定疾病期间淀粉样蛋白纤维的结构基础并发现关键的伴侣调节过程对于质量控制和清除形成的蛋白质至关重要有毒淀粉样蛋白。