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: 8606521
• 负责人: Elizabeth Rhoades
• 金额: $ 34.95万
• 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/8606521
• 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，影响约1%的65岁及以上人口。这种蛋白-突触核蛋白是路易小体的主要成分，路易小体是神经元细胞质中聚集蛋白的沉积物，是帕金森病的标志。有令人信服的证据表明，¿-synuclein聚集过程与帕金森病的病理直接相关。然而，¿-synuclein的聚集是一个复杂的、异质的过程，在这个过程中，多个低聚蛋白物种在不同的时间点聚集，并在一定的时间尺度上持续存在，这使得它的完整表征极具挑战性。此外，在聚集过程中形成的各种分子物种与疾病病理之间的关系仍然是一个悬而未决的问题，尽管寡聚-突触核蛋白与细胞膜之间的直接相互作用已被牵连。另一个复杂的问题来自最近的两篇论文，它们提供了证据，证明长期以来被认为是无序单体的¿- synuclein的天然状态实际上可能是部分结构的四聚体。我们的假设是，AS的功能和功能失调相互作用都是由平均构象或寡聚系综的变化以及这些系综内状态之间的动态交换来调节的。我们的研究将在溶液中（目标1）、与模型膜结合（目标2）和活细胞中（目标3）表征这些集合体，目的是确定与向毒性状态过渡相关的物理特征。作为这些实验的结果，我们还将确定假定的As的四聚体和单体形式之间的关系。为了做到这一点，我们将使用单分子和时间分辨荧光方法。我们的实验方法的优点是允许在有利于寡聚化或聚集的条件下表征¿-synuclein，而不需要伴随蛋白质实际自结合的信号解释的复杂性。我们的研究结果将是一个全面的观点，什么性质的溶液，膜，和细胞环境有利于¿-突触核蛋白自结合成毒性结构。这种理解对于我们研究小组确定治疗或预防帕金森病的潜在分子靶点的长期目标至关重要。