Towards the understanding of how chaperones function and prevent amyloidogenic diseases

了解伴侣如何发挥作用并预防淀粉样蛋白形成疾病

• 批准号: 10734397
• 负责人: Marielle Aulikki Hegetschweiler
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10734397
• 关键词: Adopted; Affect; Alzheimer' s Disease; American; Amyloid; Amyloid Fibrils; Amyloid beta-42; Amyloid beta-Protein; Attention; Bacteria; Binding; Binding Sites; Career Mobility; Cell Line; Cells; Communication; Cryoelectron Microscopy; Cytoprotection; Deposition; Development; Disease; Disease Progression; Encapsulated; Equilibrium; Evolution; Funding; Heat shock proteins; Human; Individual; Knowledge; Mammals; Measurement; Methods; Mitochondria; Modeling; Molecular Chaperones; Molecular Conformation; Monitor; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Organism; Peptides; Phase; Play; Prevention; Principal Investigator; Process; Proteins; Reaction; Reactive Oxygen Species; Relaxation; Research; Resolution; Role; Sampling; Site; Specificity; Stress; Structure; System; Therapeutic; Toxic effect; Training; Yeasts; abeta accumulation; beta pleated sheet; experimental study; extracellular; flexibility; insight; mitochondrial dysfunction; monomer; novel; novel therapeutic intervention; overexpression; pressure; prevent; protein aggregation; protein folding; protein misfolding; proteostasis; stressor

Project Summary/Abstract Alzheimer's disease (AD) is a fatal neurodegenerative disease affecting 5.5 million Americans. Despite many decades of research there is still no known cure. AD is a protein misfolding disease, where the Alzheimer's protein, Aβ, aggregates from a random coil entity into fibrils, which are highly organized aggregates containing a cross-β sheet structure. However, the nature of the toxic species in Alzheimer's disease remains unknown. More and more attention has been given to the possibility that Aβ aggregates within mitochondria, rather than extracellular deposits of Aβ, may be responsible for the onset and progression of the disease. Nature has developed mechanisms to prevent disease-associated protein aggregation, e.g. by the introduction of heat shock proteins (Hsp's), which are overexpressed when cells undergo stress. The most important Hsp's in mitochondria are Hsp60 and Hsp70, whereas Hsp60 is the only essential chaperone in bacteria, yeast, and mammals. It is known that Hsp60 is cytoprotective against many stressors in cells and is proposed to be directly protective against AD. However nothing is known about the mechanism of how this is achieved. Since almost nothing is known about the mitochondrial Hsp60 system, including its co-chaperone Hsp10, I will use aims 1 and 2 of this proposal to explore this chaperone in absence of substrate protein. I seek to solve the structure of different states of the chaperone during its protein folding cycle by cryo-electron microscopy (cryo-EM). Further, using novel solution-state NMR methods to study sparsely-populated states, I will investigate how the co-chaperone is involved in the reaction cycle. This will provide important information on the role of Hsp10 in substrate encapsulation. Further, during my independent phase I will use the fundament knowledge obtained through aims 1 and 2 to move on to aim 3, where I will study the interaction of Hsp60 with Aβ. I will not only elucidate the structure of long-lived major states of Aβ bound to Hsp60 after substrate encapsulation by cryo-EM, but also study the transient interaction of Aβ with Hsp60 during the initial phase of the substrate folding cycle, prior to encapsulation, by solution-state NMR. These studies will provide information about which Aβ species (monomer, oligomer, or fibril) interact with Hsp60 and elucidate their structural features. Aims 1 and 2 will provide insights into fundamental questions about the mechanisms chaperones use to efficiently fold proteins into their functional forms. Further the results from aim 3 will unveil details about how Hsp60 inhibits Aβ aggregation and prevents neurodegenerative diseases, and may open up novel therapeutic strategies against Alzheimer's disease.

项目摘要/摘要 阿尔茨海默氏病（AD）是一种致命的神经退行性疾病，影响了550万美国人。尽管 数十年的研究仍然没有已知的治愈方法。 AD是一种蛋白质错误折叠疾病，其中 阿尔茨海默氏症的蛋白质Aβ，从随机线圈实体聚集到原纤维，这些纤维是高度组织的 包含跨β结构的聚集体。但是，阿尔茨海默氏症中有毒物种的性质 疾病仍然未知。越来越关注Aβ聚集物的可能性 在线粒体内，而不是Aβ的细胞外沉积物，可能是造成发作和 疾病的进展。大自然已经开发了预防疾病相关蛋白的机制 聚集，例如通过引入热激蛋白（HSP），当细胞时会过表达 承受压力。线粒体中最重要的HSP是HSP60和HSP70，而HSP60是 仅细菌，酵母和哺乳动物中的必需伴侣。众所周知，HSP60对 细胞中的许多压力源，并被提议直接保护AD。但是什么都不知道 关于如何实现的机制。 由于线粒体HSP60系统几乎一无所知，包括其副酮HSP10，我 将使用该提案的目标1和2在没有底物蛋白质的情况下探索该伴侣。我寻求 通过冷冻电子蛋白质折叠循环解决伴侣蛋白不同状态的结构 显微镜（冷冻EM）。此外，使用新型的溶液状态NMR方法研究稀少 国家，我将研究辅助酮如何参与反应周期。这将提供重要的 有关HSP10在底物封装中的作用的信息。此外，在我的独立阶段，我将 利用通过目标1和2获得的基础知识继续前进到AIM 3，我将在其中研究 Hsp60与Aβ的相互作用。我不仅会阐明Aβ长期寿命的主要状态的结构 HSP60在用冷冻EM封装后的HSP60，但也研究Aβ与Hsp60的瞬时相互作用 在封装之前的基材折叠周期的初始阶段，通过溶液状态NMR。这些 研究将提供有关哪种Aβ物种（单体，低聚物或原纤维）与HSP60相互作用的信息 并阐明其结构特征。目标1和2将提供有关有关的基本问题的见解 伴侣使用的机制可以有效地将蛋白质折叠成其功能形式。进一步的结果 从AIM 3提出有关HSP60如何抑制Aβ聚集并防止神经退行性的细节 疾病，并可能开辟针对阿尔茨海默氏病的新型治疗策略。