PROBING ALPHA-SYNUCLEIN AGGREGATION

探测 α-突触核蛋白聚集

• 批准号: 8364109
• 负责人: ELKA R GEORGIEVA
• 金额: $ 0.99万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364109
• 关键词: Amyloid Fibrils; Behavior; Binding; Cysteine; Cytosol; Development; Disease; Dopamine; Environment; Funding; Goals; Grant; Homeostasis; Learning; Lewy Bodies; Link; Maintenance; Membrane; Membrane Proteins; Micelles; Molecular Conformation; National Center for Research Resources; Parkinson Disease; Phospholipids; Physiologic pulse; Point Mutation; Principal Investigator; Property; Proteins; Regulation; Research; Research Infrastructure; Resources; Source; Spin Labels; Structure; Synapses; Technology; United States National Institutes of Health; Variant; alpha synuclein; alpha synuclein gene; cost; mimetics; mutant; presynaptic; protein aggregation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Alpha-synuclein (aS) is a highly conserved presynaptic protein that participates in synaptic strength maintenance and dopamine homeostasis. It is also involved in regulation of intracellular dopamine levels at several points of control. However, accumulation of aS amyloid fibrils was implicated as the hallmark in the development of Parkinson's disease. The three single-point mutations in a-synuclein, namely, A30P, A53T, and E46K, (as well as a triplication of the aS gene) have been linked to a rare familial form of Parkinson's disease (PD). On the other hand the vast majority of Lewy body-related disease cases is sporadic and involves the wild type of aS. Normal functions of ¿S involve protein-membrane interactions upon which the protein undergoes transformations from a disordered structure in cytosol to the highly helical one in membrane-bound state. In several completed subprojects we have successfully applied Pulsed dipolar ESR to characterize the behavior of wild type aS bound to SDS micelles and phospholipid membranes [1, 2]. In our detailed study on the structural properties of WT and PD-linked mutants A30P, E46K and A53, we showed that all variants of aS posses the propensity to switch between broken and extended helix conformations, depending on aS environment [1-3]. Yet, there is insufficient information about the factors that trigger and govern the aggregation of this protein. Therefore, we undertook a study that has as its goal to learn whether and how the interaction with the common membrane mimetics, such as SDS, could initiate the aggregation of aS. So far, we have studied the interaction with SDS of single spin-labeled cysteine mutants of aS. [1] Borbat, P.; Ramlall, T. F.; Freed, J. H.; Eliezer, D. J. Am. Chem. Soc., 2006, 128, 10004-10005. [2] Elka R. Georgieva, Trudy F. Ramlall, Peter P. Borbat, Jack H. Freed, David Eliezer, J. Am. Chem. Soc. 2008, 130, 12856-12857. [3] E.R. Georgieva, T.F. Ramlall, P.P. Borbat, J.H. Freed, D. Eliezer, J Biol Chem, 285 (2010) 28261-28274.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供 该子项目的主要支持。 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源的子项目可能列出的总成本。 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 α-突触核蛋白 (aS) 是一种高度保守的突触前蛋白，参与突触强度维持和多巴胺稳态，它还参与多个控制点的细胞内多巴胺水平的调节，但 aS 淀粉样原纤维的积累被认为是其标志。 α-突触核蛋白的三个单点突变，即 A30P、A53T 和E46K（以及 aS 基因的三倍体）与一种罕见的家族性帕金森病 (PD) 有关，另一方面，绝大多数路易体相关疾病病例是散发性的，且涉及野生型帕金森病。 aS 的正常功能。 S 蛋白-膜相互作用，蛋白质从胞质中的无序结构转变为膜结合状态的高度螺旋结构。在几个已完成的子项目中，我们已成功应用脉冲偶极 ESR 来表征与 SDS 结合的野生型 aS 的行为。胶束和磷脂膜 [1, 2]。 在我们对 WT 和 PD 连锁突变体 A30P、E46K 和 A53 结构特性的详细研究中，我们发现 aS 的所有变体都具有在断裂和延伸螺旋构象之间转换的倾向，具体取决于 aS 环境 [1-3]。然而，关于触发和控制这种蛋白质聚集的因素的信息还不够，因此，我们进行了一项研究，其目标是了解是否以及如何与普通膜相互作用。模拟物，例如 SDS，可以启动 aS 的聚集。 到目前为止，我们已经研究了 aS 的单个自旋标记半胱氨酸突变体与 SDS 的相互作用。 ；Eliezer，D.J.化学，2006，128，10004-10005。 R. Georgieva，Trudy F. Ramlall，Peter P. Borbat，Jack H. Freed，J. Am Chem，2008，130，12856-12857。 J.H.弗里德，D.埃利泽，生物化学杂志，285 (2010)28261-28274。