Self-association and membrane-binding of alpha-synuclein

α-突触核蛋白的自缔合和膜结合

• 批准号: 8506418
• 负责人: Elizabeth Rhoades
• 金额: $ 35.3万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8506418
• 关键词: Acetylcysteine; Acidic Region; Affect; Binding; C-terminal; Cartoons; Cell Death; Cells; Cellular Membrane; Cereals; Characteristics; Complex; Complication; Consensus; Defect; Deposition; Development; Disease; Dopaminergic Cell; Drosophila genus; Energy Transfer; Environment; Fluorescence; Fluorescence Resonance Energy Transfer; Functional disorder; Genes; Goals; Investigation; Label; Lewy Bodies; Life; Link; Lipid Bilayers; Lipids; Measurement; Measures; Membrane; Methods; Modeling; Molecular; Molecular Target; Motor; Movement Disorders; Mutation; N-terminal; Nerve Degeneration; Neurons; Parkinson Disease; Pathology; Point Mutation; Population; Positioning Attribute; Process; Property; Proteins; Publications; Research; Rodent; Role; Signal Transduction; Solutions; Spectrum Analysis; Structure; Synaptic Vesicles; Time; Toxic effect; Transgenic Organisms; Work; aged; alpha synuclein; insight; membrane model; monomer; mutant; novel; overexpression; prevent; protein aggregate; public health relevance; research study; single molecule; synuclein; therapeutic development

DESCRIPTION (provided by applicant): Parkinson's disease is the most common neurodegenerative movement disorder, affecting an estimated 1% of the population aged 65 and older. The protein ¿-synuclein is the primary component of Lewy bodies, the neuronal cytoplasmic deposits of aggregated proteins that are the hallmark of Parkinson's disease. There is compelling evidence that the process of ¿-synuclein aggregation is directly related to Parkinson's disease pathology. However, the aggregation of ¿-synuclein is a complex, heterogeneous process during which multiple oligomeric protein species are populated at various timepoints and persist over a range of timescales, making its complete characterization extremely challenging. Moreover, the relationship between the various molecular species formed in the aggregation process and disease pathology is still an open question, although the direct interaction between oligomeric ¿-synuclein and cellular membranes has been implicated. An additional complication comes from two recent publications that provide evidence that the native state of ¿- synuclein, long believed to be a disordered monomer, may actually be a partially structured tetramer. Our hypothesis is that both the functional and dysfunctional interactions of AS are modulated by changes in the average conformational or oligomeric ensemble and the dynamic interchange between states within these ensembles. Our research will characterize these ensembles in solution (Aim 1), bound to model membranes (Aim 2), and in living cells (Aim 3), with the goal of determining physical features that are associated with th transition to toxic states. As a consequence of these experiments, we will also determine the relationship between the putative tetramer and monomer forms of AS. To do this, we will use single molecule and time- resolved fluorescence methods. Our experimental approaches have the advantage of allowing for the characterization of ¿-synuclein under conditions that favor oligomerization or aggregation without the complication of signal interpretation that accompanies actual self-association of the protein. The results of our investigation will be a comprehensive view of what properties of the solution, membrane, and cellular environment favor self-association of ¿-synuclein into toxic structures. Such understanding is critical to the long term goal of our research group to identify potential molecular targets for the development of therapeutics to treat or prevent Parkinson's disease.

描述（由申请人提供）：帕金森病是最常见的神经退行性运动障碍，估计影响 1% 的 65 岁及以上人口。 -突触核蛋白是路易体的主要成分，路易体是聚集蛋白的神经元细胞质沉积物，是帕金森氏病的标志。 -突触核蛋白聚集与帕金森病病理学直接相关，然而，¿突触核蛋白是一个复杂的异质过程，在此过程中多种寡聚蛋白种类在不同的时间点聚集并在一系列时间尺度上持续存在，这使得其完整表征极具挑战性。此外，在聚集过程中形成的各种分子种类与疾病之间的关系极具挑战性。尽管寡聚体之间存在直接相互作用，但病理学仍然是一个悬而未决的问题。 -突触核蛋白和细胞膜也被牵连，最近的两篇出版物提供了 ¿ 天然状态的证据。 - 突触核蛋白，长期以来被认为是无序单体，实际上可能是部分结构化的四聚体。我们的假设是，AS 的功能和功能障碍相互作用都是通过平均构象或寡聚体整体的变化以及这些状态之间的动态交换来调节的。我们的研究将表征这些集合在溶液中（目标 1）、与模型膜（目标 2）和活细胞（目标 3）的结合，目的是确定与向细胞膜过渡相关的物理特征。作为这些实验的结果，我们还将确定 AS 的假定四聚体和单体形式之间的关系。为此，我们将使用单分子和时间分辨荧光方法。用于表征 ¿ -在有利于寡聚或聚集的条件下，没有伴随蛋白质实际自缔合的信号解释的复杂性，我们的研究结果将全面了解溶液、膜和细胞环境的哪些特性有利于自缔合。的协会这种理解对于我们研究小组的长期目标至关重要，即确定开发治疗或预防帕金森病的疗法的潜在分子靶标。