Structures and biological activity of alpha-synuclein aggregation

α-突触核蛋白聚集的结构和生物活性

• 批准号: 10390302
• 负责人: Lin Jiang
• 金额: $ 31.1万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390302
• 关键词: Address; Affect; Amyloid; Amyloid Fibrils; Amyloid Proteins; Biological; Brain; Cells; Cellular Assay; Computer software; Cost Measures; Cryoelectron Microscopy; Data; Disease; Economic Burden; Electrons; Elements; Ensure; Familial disease; Fiber; Future; Genetic Polymorphism; Goals; In Vitro; Length; Lewy Body Disease; Methods; Modeling; Molecular; Molecular Conformation; Monitor; Multiple System Atrophy; Mutation; Neurodegenerative Disorders; Parkinson Disease; Pathologic; Patients; Polymorph; Preparation; Procedures; Process; Proteins; Public Health; Recombinants; Research; Resolution; Sampling; Seeds; Source; Structural Models; Structure; Structure-Activity Relationship; Technology; Toxic effect; Work; alpha synuclein; cytotoxicity; drug development; effective therapy; experimental study; in vivo; mutant; new therapeutic target; preservation; success; synucleinopathy; therapeutic target

Summary Synucleinopathies are a group of neurodegenerative disorders that have been associated to the misfolded amyloid protein α-synuclein in brain. The misfolded α-synuclein can aggregate into polymorphic fibrils, displaying distinct biological activities and contributing differently to the diseases. However, the underlying mechanisms remain unclear. To address this, we will integrate both structural and functional approaches to study recombinant wild type, disease-related mutants, and brain-derived α-synuclein fibrils. First, we will use cryo-Electron Microscopy (cryo-EM), combined with other structural methods, to achieve near-atomic structures of recombinant full-length fibrils of wild type and disease mutant α-synuclein (Aims 1-2). We will then go beyond structure determination and utilize cellular assays of seeding and toxicity to explore the biological activities of both wild type and disease mutant α-synuclein fibrils. By comparing the structures of these fibrils, we will determine structure elements responsible for their biological activity, and correlate structural differences to seeding and toxicity for a better understanding of their structure-activity relationship (Aim 2). Finally, we will determine cryo-EM structures of in vivo α-synuclein fibrils derived from the patient brains of synucleinopathies. Thus we can assess the pathological relevance of the determined recombinant fibril structures and relate structural features observed in α-synuclein fibrils to different disease states (Aim 3). Our integrated approach, connecting structure and biological activity of α-synuclein fibrils of different forms or sources, will reveal atomic understanding of the underlying mechanisms and provide therapeutic targets suitable for future drug development that precisely targets α-synuclein aggregation to stop synucleinopathies.

总结 突触核蛋白病是一组神经退行性疾病， 淀粉样蛋白α-突触核蛋白。错误折叠的α-突触核蛋白可以聚集成多形原纤维， 表现出不同的生物活性，并对疾病有不同的贡献。但是，底层 其机制仍不清楚。为了解决这个问题，我们将整合结构和功能方法， 研究重组野生型、疾病相关突变体和脑源性α-突触核蛋白原纤维。首先，我们将使用 冷冻电子显微镜（cryo-EM），结合其他结构方法，以实现近原子 野生型和疾病突变体α-突触核蛋白的重组全长原纤维的结构（目的1-2）。我们将 然后超越结构测定，利用接种和毒性的细胞测定来探索 野生型和疾病突变体α-突触核蛋白原纤维的生物活性。通过比较 这些纤维，我们将确定负责其生物活性的结构元素，并将其与 接种和毒性的结构差异，以便更好地了解它们的结构-活性关系 (Aim 2）的情况。最后，我们将确定来自患者的体内α-突触核蛋白原纤维的冷冻电镜结构 突触核蛋白病的大脑因此，我们可以评估确定的重组的病理相关性， 纤维结构和相关的结构特征在α-突触核蛋白纤维中观察到不同的疾病状态（目的3）。 我们的综合方法，连接结构和生物活性的α-突触核蛋白纤维的不同形式或 将揭示对潜在机制的原子理解，并提供治疗靶点。 适用于未来精确靶向α-突触核蛋白聚集以阻止突触核蛋白病的药物开发。