Understanding the Effect of N-Terminal Acetylation on alpha-Synuclein Toxicity

了解 N 末端乙酰化对 α-突触核蛋白毒性的影响

• 批准号: 8455155
• 负责人: Jennifer Megan Beierlein
• 金额: $ 4.92万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-16 至 2014-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8455155
• 关键词: Acetylation; Alzheimer' s Disease; Amyloid; Biochemical; Biological Assay; Biology; Biomolecular Nuclear Magnetic Resonance; Categories; Cells; Circular Dichroism; Clinical Markers; Color; Congo Red; Disease; Drug Delivery Systems; Drug Design; Electron Microscopy; Escherichia coli; Genes; Health; Human; In Vitro; Kinetics; Label; Microscopy; Modification; Movement Disorders; Mutation; N-terminal; NMR Spectroscopy; Nerve Degeneration; Neurodegenerative Disorders; Parkin gene; Parkinson Disease; Pathogenesis; Play; Post-Translational Protein Processing; Proteins; Role; Saccharomyces cerevisiae; Site; Staining method; Stains; Structure; Substantia nigra structure; System; Thioflavin T; Toxic effect; Yeast Model System; Yeasts; alpha synuclein; amyloid formation; base; design; improved; in vivo; insight; interest; monomer; mutant; overexpression; parkin gene/protein; protein function; research study; synuclein; synucleinopathy; yeast genetics

DESCRIPTION (provided by applicant): Parkinson's Disease (PD) is a debilitating neurodegenerative movement disorder, whose clinical markers include the formation of alpha-Synuclein (alpha-Syn) rich aggregates in the substantia nigra . Despite the identification of alpha-Syn, a 14 kDa protein, as a target of interest in the treatment of PD, much remains unknown about this protein. Factors behind its aggregation and toxicity are unclear, however studies have indicated that posttranslational modifications play an important role. N-terminal acetylation has been seen as a potential factor in the disease state, yet this modification remains largely unstudied. This project is designed to clarify the role of N- terminal acetylation and the effects of this modification on alpha-Syn. The stability and toxicity of the modified and unmodified proteins will be determined through in vivo assays in yeast studying the aggregation and subcellular localization of alpha-Syn. Structural studies utilizing circular dichroism and NMR spectroscopy will examine any structural changes resulting from the posttranslational modifications. The effect of these posttranslational modifications on the PD-associated mutants, A30P, E46K, and A53T, will also be studied. This project will provide insight into the cause of alpha-Syn toxicity, aiding structure-based drug design efforts. PUBLIC HEALTH RELEVANCE: Parkinson's disease, a debilitating movement disorder, is the second most prevalent neurodegenerative disease after Alzheimer's disease(1). Aggregation of a 14 kDa protein, alpha-Synuclein, has been identified as a histopathological hallmark of the disease, however many questions remain that hinder drug design efforts. This project is designed to further probe the structure, stability and toxicity of alpha-Synuclein, improving understanding of this important drug target.

描述（由申请人提供）：帕金森病（PD）是一种使人衰弱的神经退行性运动障碍，其临床标志包括黑质中富含α-突触核蛋白（α-Syn）的聚集体的形成。尽管 α-Syn（一种 14 kDa 蛋白质）已被鉴定为帕金森病治疗的靶标，但对该蛋白质仍有许多未知之处。其聚集和毒性背后的因素尚不清楚，但研究表明翻译后修饰发挥着重要作用。 N 末端乙酰化已被视为疾病状态的潜在因素，但这种修饰在很大程度上仍未得到研究。该项目旨在阐明 N 末端乙酰化的作用以及这种修饰对 α-Syn 的影响。修饰和未修饰蛋白质的稳定性和毒性将通过研究 α-Syn 聚集和亚细胞定位的酵母体内测定来确定。利用圆二色性和核磁共振波谱的结构研究将检查翻译后修饰引起的任何结构变化。还将研究这些翻译后修饰对 PD 相关突变体 A30P、E46K 和 A53T 的影响。该项目将深入了解 α-Syn 毒性的原因，有助于基于结构的药物设计工作。 公众健康相关性：帕金森病是一种使人衰弱的运动障碍，是仅次于阿尔茨海默病的第二大常见神经退行性疾病(1)。 14 kDa 蛋白质 α-突触核蛋白的聚集已被确定为该疾病的组织病理学标志，但仍有许多问题阻碍了药物设计工作。该项目旨在进一步探讨α-突触核蛋白的结构、稳定性和毒性，增进对这一重要药物靶点的了解。