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: 8545599
• 负责人: Jennifer Megan Beierlein
• 金额: $ 5.02万
• 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 2014-09-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8545599
• 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 Design; Drug Targeting; 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.

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，其临床标志物包括黑质中富含α-突触核蛋白（α-Syn）的聚集体的形成。尽管鉴定了α-Syn（一种14 kDa蛋白质）作为治疗PD的目标靶标，但关于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，翻译后修饰起着重要作用。N-末端乙酰化已被视为疾病状态中的潜在因素，但这种修饰在很大程度上尚未研究。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。修饰和未修饰的蛋白质的稳定性和毒性将通过在酵母中研究α-Syn的聚集和亚细胞定位的体内测定来确定。利用圆二色性和NMR光谱的结构研究将检查由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A30 P、E46 K和A53 T的影响。该项目将深入了解alpha-Syn毒性的原因，帮助基于结构的药物设计工作。