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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 α-突触核蛋白（AS）是一种高度组成的突触前蛋白，参与突触强度维持和多巴胺稳态。它还参与了几个控制点的细胞内多巴胺水平的调节。但是，淀粉样蛋白原纤维的积累被作为帕金森氏病发展的标志。 A-突触核蛋白的三个单点突变，即A30p，A53T和E46K（以及AS基因的三重）与罕见的农场形式的帕金森氏病（PD）有关。另一方面，绝大多数与Lewy身体相关的疾病病例都是零星的，涉及AS的野生类型。 S的正常功能涉及蛋白质 - 膜相互作用，蛋白质从细胞质中的无序结构转变为膜结合状态的高度螺旋形的结构。在几个完整的子项目中，我们成功地应用了脉冲偶极ESR，以表征与SDS胶束和磷脂膜结合的野生类型的行为[1，2]。 在我们对WT和PD连接突变体A30P，E46K和A53的结构特性的详细研究中，我们表明，所有AS的变体都构成了在损坏和扩展的螺旋组成之间切换的希望，这取决于AS环境[1-3]。然而，关于触发和控制该蛋白质聚集的因素的信息不足。因此，我们进行了一项研究，该研究的目标是了解与常见膜模拟物（例如SDS）的相互作用如何以及如何启动AS的聚集。到目前为止，我们已经研究了与AS的单个自旋标记半胱氨酸突变体SD的相互作用。 [1] Borbat，P。; Ramlall，T。F。; Freed，J。H。; Eliezer，D。J. Am。化学Soc。，2006，128，10004-10005。 [2] Elka R. Georgieva，Trudy F. Ramlall，Peter P. Borbat，Jack H. Freed，David Eliezer，J。Am。化学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。