Engineering Ubiquitin Ligases to Investigate Protein Aggregation and Neurodegener

工程泛素连接来研究蛋白质聚集和神经变性

• 批准号: 7847875
• 负责人: Shu-Bing Qian
• 金额: $ 231万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7847875
• 关键词: Affect; Age; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Binding; Biological Process; Brain; Cell Cycle Regulation; Cells; Dementia; Disease; Engineering; Functional disorder; Goals; Health; Heart; Huntington Disease; Knock-out; Ligase; Modeling; Molecular; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Parkinson Disease; Patients; Population; Prevalence; Proteins; Proteomics; Public Health; Quality Control; Recombinants; Research; Role; Signal Transduction; Somatic Cell; Specificity; System; Therapeutic; Ubiquitin; Ubiquitination; abstracting; human Huntingtin protein; human disease; improved; innovation; insight; method development; multicatalytic endopeptidase complex; novel; novel strategies; protein aggregate; protein aggregation; protein degradation; protein function; protein misfolding; prototype; public health relevance; tool; ubiquitin ligase

DESCRIPTION (Provided by the applicant) Abstract: Protein degradation lies at the heart of biological processes from signal transduction to cell cycle regulation. Compromised clearance of misfolded proteins from cells is the leading cause of human diseases such as neurodegenerative disorders. A common theme manifested in neurodegeneration is the accumulation of insoluble protein aggregates in the brain. However, the role of protein aggregation in the pathophysiology of neurodegeneration remains controversial. It remains a formidable task to remove protein aggregates from the affected neurons. In this proposal, we are taking a bold and innovative approach to attacking this exceedingly difficult problem: harnessing the ubiquitin/proteasome system to investigate protein aggregation and neurodegeneration. This proposal builds upon our previous development of methods to engineer single chain ubiquitin ligase CHIP. We successfully established a novel strategy that enables us to alter the substrate binding specificity of CHIP without affecting its ligase activity. Using Huntington's disease (HD) as a model, we propose herein to create recombinant ubiquitin ligases targeting the disease protein huntingtin (Htt) for ubiquitination. Our long-term goal is to define the structural features that determine the ligase activity of engineered ubiquitin ligases, elucidate the molecular mechanisms underlying protein aggregation and neurodegeneration, and evaluate the therapeutic potential of Htt-specific ubiquitin ligases. If successful, the engineered ubiquitin ligases will represent an unprecedented level of control over protein function in somatic cells, which would have direct impact on proteomic research by introducing novel "protein knockout" tools. In addition, the results of this project will provide unique insights into the fundamental cellular and molecular mechanisms underlying protein quality control and the pathophysiology of neurodegeneration. Application of these findings may help to delay or reverse the detrimental effects of neurodegeneration. Ultimately, it will serve as a prototype for the treatment of other neurodegenerative disorders, as well as non-neuronal human diseases. Public Health Relevance: As our population continues to age, neurodegenerative diseases will increase in prevalence and thus pose a daunting challenge to public health worldwide. These truly disastrous disorders include Alzheimer's, Huntington's, Parkinson's, amyotrophic lateral sclerosis and the frontal temporal dementias. This proposal aims to develop novel tools to target the culprits of neurodegeneration, elucidate underlying mechanisms, and provide therapeutic potential to improve the health of these patients.

描述（由申请人提供） 翻译后摘要：蛋白质降解的核心是从信号转导到细胞周期调控的生物过程。错误折叠蛋白质从细胞中的受损清除是人类疾病如神经退行性疾病的主要原因。在神经变性中表现的一个共同主题是不溶性蛋白质聚集体在脑中的积累。然而，蛋白质聚集在神经变性的病理生理学中的作用仍然存在争议。从受影响的神经元中去除蛋白质聚集体仍然是一项艰巨的任务。在这项提案中，我们采取了一种大胆而创新的方法来解决这个极其困难的问题：利用泛素/蛋白酶体系统来研究蛋白质聚集和神经变性。这个建议建立在我们以前开发的方法工程单链泛素连接酶CHIP。我们成功地建立了一种新的策略，使我们能够改变CHIP的底物结合特异性，而不影响其连接酶活性。使用亨廷顿氏病（HD）作为模型，我们在此提出创建靶向疾病蛋白亨廷顿蛋白（Htt）的重组泛素连接酶用于泛素化。我们的长期目标是确定的结构特征，确定工程泛素连接酶的连接酶活性，阐明蛋白质聚集和神经变性的分子机制，并评估的治疗潜力的Htt特异性泛素连接酶。如果成功，工程泛素连接酶将代表对体细胞中蛋白质功能的前所未有的控制水平，这将通过引入新的“蛋白质敲除”工具对蛋白质组学研究产生直接影响。此外，该项目的结果将为蛋白质质量控制和神经变性的病理生理学基础的基本细胞和分子机制提供独特的见解。这些发现的应用可能有助于延缓或逆转神经退行性变的有害影响。最终，它将成为治疗其他神经退行性疾病以及非神经元人类疾病的原型。 公共卫生相关性：随着人口的不断老龄化，神经退行性疾病的患病率将增加，从而对全球公共卫生构成严峻挑战。这些真正灾难性的疾病包括阿尔茨海默氏症、亨廷顿氏症、帕金森氏症、肌萎缩侧索硬化症和额颞痴呆症。该提案旨在开发新的工具来靶向神经退行性变的罪魁祸首，阐明潜在的机制，并提供改善这些患者健康的治疗潜力。