NMR Based Studies of Alpha-Synuclein Aggregation and Inhibition

基于 NMR 的 α-突触核蛋白聚集和抑制研究

• 批准号: 9527263
• 负责人: JEAN S BAUM
• 金额: $ 4.7万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9527263
• 关键词: Address; Affect; Amyloid; Amyloid beta-Protein; Architecture; Binding; Biological Assay; Brain; Cell model; Cells; Characteristics; Chimera organism; Complement; Complex; Cysteine; Data; Dependence; Development; Dimerization; Disease; Dissociation; Disulfides; Drug Design; Goals; Grant; Homo; Incubated; Kinetics; Laboratories; Light; Link; Location; Maps; Modeling; Molecular; Molecular Conformation; Monitor; N-terminal; Nature; Neurodegenerative Disorders; Neurons; Outcome; Parkinson Disease; Pathologic; Patients; Pharmacotherapy; Play; Process; Property; Protein Precursors; Proteins; Recruitment Activity; Role; Seeds; Site; Specificity; Structure; Testing; Therapeutic Intervention; To specify; Toxic effect; Work; aging population; alpha synuclein; base; biophysical properties; biophysical techniques; crosslink; cytotoxic; design; dimer; driving force; experimental study; fighting; global health; in vivo; inhibitor/antagonist; insight; intermolecular interaction; monomer; mutant; new therapeutic target; novel; preference; prevent; public health relevance; synuclein; tau Proteins

 DESCRIPTION (provided by applicant): The mechanism of aggregation in neurodegenerative disease has been investigated by numerous laboratories and although the process is still elusive, significant progress has been made in defining different stages of the process. To fight these debilitating illnesses it is not only critical to characterize aggregation processes but alsoto understand the mechanisms of aggregation inhibition. N-terminally acetylated a-synuclein (Ac-aS) is a small neuronal intrinsically disordered protein (IDP) that self-associates in pathological ways to oligomers and fibrils in the brains of patients with Parkinson's disease. In contrast, a highly homologous IDP protein, acetylated ß- synuclein (Ac-ßS) that co-localizes with Ac-aS, does not self-associate into fibrils but can act as a neuro-protector of Ac-aS toxicity in vivo to inhibit pathological Ac-aS fibrillation. In light of the incredible importance of neurodegenerative diseases and their threat to global health in the ageing population, a detailed understanding of the molecular mechanism of inhibition is critical. At this stage very little known about these processes and about the specific inter-molecular interactions that guide inhibition versus aggregation. The goal of this proposal is to use NMR and other biophysical techniques, in conjunction with cell toxicity studies, to provide the first molecular description of the complexes that exist at the different stages of inhibition. In this grant we will: (1) characterize the influnce of different aS/ßS sequences, or structural units, on the potential to accelerate/inhibit fibril formation by the natural inhibitor ßS and its toxic mutants; (2) map interactive regions that define the earliest stages of inhibition; and (3) undertake the molecular characterization and toxicity studies of aS, ßS and hetero aS/ßS oligomers. The outcome of the proposed work will be to advance our fundamental understanding of the 'structure-toxicity' relationship of aS/ßS inhibition and to address more global questions about the location, nature and specificity of inhibitory interactions. These studies will provide a framework for identifying specific interactios that are important for disrupting toxic oligomers and for proposing novel targets for therapeutic intervention in Parkinson's disease. The approaches developed here can be more widely applied to other cross amyloid interactions that have been shown to play a critical role in neurodegenerative disease, such as the interaction between aS and amyloid-ß-protein, or aS and tau.

 描述（由申请人提供）：许多实验室已经研究了神经退行性疾病中的聚集机制，尽管该过程仍然难以捉摸，但在定义该过程的不同阶段方面已经取得了重大进展。为了对抗这些使人衰弱的疾病，不仅要描述聚集过程的特征，而且要了解聚集抑制的机制。 N 末端乙酰化 a-突触核蛋白 (Ac-aS) 是一种小型神经元本质紊乱蛋白 (IDP)，在病理状态下自关联 帕金森病患者大脑中寡聚体和原纤维的方法。相比之下，高度同源的 IDP 蛋白乙酰化 ß-突触核蛋白 (Ac-ßS) 与 Ac-aS 共定位，不会自行结合成原纤维，但可以作为体内 Ac-aS 毒性的神经保护剂，抑制病理性 Ac-aS 纤维颤动。鉴于神经退行性疾病的重要性 在老龄化人口中，疾病及其对全球健康的威胁，详细了解抑制的分子机制至关重要。在这个阶段，人们对这些过程以及指导抑制与聚集的特定分子间相互作用知之甚少。该提案的目标是使用核磁共振和其他生物物理技术，结合细胞毒性研究，提供复合物的第一个分子描述 存在于抑制的不同阶段。在这笔资助中，我们将：（1）表征不同的 aS/ßS 序列或结构单元对天然抑制剂 ßS 及其有毒突变体加速/抑制原纤维形成的潜力的影响； (2) 绘制定义抑制最早阶段的交互区域； (3) 进行 aS、ßS 和异质 aS/ßS 寡聚物的分子表征和毒性研究。拟议工作的结果将是增进我们对 aS/ßS 抑制的“结构-毒性”关系的基本理解，并解决有关抑制相互作用的位置、性质和特异性的更多全球性问题。这些研究将为识别特定的相互作用提供一个框架，这些相互作用对于破坏有毒寡聚体和提出帕金森病治疗干预的新靶标非常重要。这里开发的方法可以更广泛地应用于其他跨淀粉样蛋白相互作用，这些相互作用已被证明在神经退行性疾病中发挥着关键作用，例如 aS 和淀粉样蛋白-β-蛋白之间的相互作用，或 aS 和 tau 之间的相互作用。