THE MOLECULAR BASIS OF ALPHA-SYNUCLEIN AGGREGATION

描述(摘自申请人的摘要)：最近，α-突触核蛋白已经 被鉴定为路易小体的主要成分，细胞内包裹体 这些都是帕金森氏病(PD)的病理特征。我们在这方面的目标 是为了检验帕金森氏症的关键一步 是α-突触核蛋白的聚集，这导致了路易的形成 身体和随后的神经元死亡。具体来说，我们将确定 α-突触核蛋白聚集的分子基础和研究潜力 α-粘连蛋白聚集的抑制剂。 我们的初步结果揭示了一些因素，导致 中性pH条件下α-突触核蛋白的构象变化和聚集 和原纤维形成。我们计划对生物物理学进行系统的描述 确定α-突触核蛋白的性质是否与 它的构象和聚集倾向，既有野生型，也有野生型 突变型α-突触核蛋白。我们将调查各种因素是否与 对于钯，例如金属离子和杀虫剂，增强了聚集性 α-突触核蛋白。将研究汇总过程的细节，以 阐明了聚集和原纤维形成的分子机制。我们会 筛选一系列多肽和小分子对小鼠胸腺细胞的抑制作用 α-突触核蛋白聚集。 这些实验代表着为阐明其作用的关键步骤。 帕金森氏病中的α-突触核蛋白。我们希望了解到潜在的角色 各种因素，从环境污染物到 触发纤维形成的α-突触核蛋白的浓度。此外，我们 预期Fnding抑制剂可能为潜在的 治疗方法。要使用的技术包括各种 生物物理/生化方法，如衰减全反射傅里叶变换红外光谱 分析聚集的α-突触核蛋白、原子力的构象状态 和电子显微镜成像聚集体，和动力学方法监测 纤维形成的速度。

DESCRIPTION (From the applicant's abstract): Recently,alpha-synuclein has been identified as a major component of Lewy bodies, the intracellular inclusions that are a pathological hallmark of Parkinson's disease (PD). Our goals in this proposal are to test the hypothesis that a critical step in Parkinson's disease is the aggregation of alpha-synuclein, which leads to the formation of Lewy Bodies and subsequently to neuronal death. Specifically we will determine the molecular basis for alpha-synuclein aggregation and investigate potential inhibitors of alpha-syouclein aggregation. Our preliminary results have revealed a number of factors that lead to a confonnational change in alpha synuclein at neutral pH, and also to aggregation and fibril formation. We plan a systematic characterization of the biophysical properties of alpha-synuclein to determine if there is a correlation between its conformation and its propensity to aggregate, with both wild type and mutant alpha-synucleins. We will investigate whether various factors associated with PD, for example, metal ions and pesticides, enhance the aggregation of alpha-synuclein. Details of the aggregation process will be studied to elucidate the molecular mechanism of aggregation and fibril formation. We will screen a series of peptides and small molecules for inhibitory effects on alpha-synuclein aggregation. These experiments represent critical steps towards elucidating the role of alpha-synuclein in Parkinson's disease. We expect to learn the potential role of various factors, ranging from environmental contaminants to the concentration of alpha-synuclein, in triggering fibril formation. Further, we anticipate fnding inhibitors which may lay the groundwork for potential therapeutic approaches. Techniques to be used include various biophysical/biochemical methods, such as attenuated total reflectance FTIR to analyze the conformaffonal state of aggregated alpha-synuclein, atomic force and electron microscopy to image the aggregates, and kinetic methods to monitor the rate of formation of fibrils.