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

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，其临床标志包括在黑质中形成富含α -突触核蛋白（α - syn）的聚集体。尽管已经鉴定出α - syn（一种14kda的蛋白）作为PD治疗的靶标，但对该蛋白仍有许多未知之处。其聚集和毒性背后的因素尚不清楚，但研究表明，翻译后修饰起重要作用。n端乙酰化被认为是疾病状态的一个潜在因素，但这种修饰在很大程度上仍未被研究。本项目旨在阐明N端乙酰化的作用和这种修饰对α - syn的影响。经过修饰和未修饰的蛋白的稳定性和毒性将通过在酵母体内研究α - syn的聚集和亚细胞定位来确定。利用圆二色性和核磁共振光谱的结构研究将检查翻译后修饰引起的任何结构变化。这些翻译后修饰对pd相关突变体A30P、E46K和A53T的影响也将被研究。该项目将深入了解α - syn毒性的原因，帮助基于结构的药物设计工作。

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.