COPPER BINDING TO ALPHA-SYNUCLEIN, THE PARKINSON'S PROTEIN

铜与帕金森病蛋白 α-突触核蛋白的结合

• 批准号: 8170119
• 负责人: SERENA D DE BEER GEORGE
• 金额: $ 0.44万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170119
• 关键词: Affinity; Binding; Binding Sites; Chemistry; Computer Retrieval of Information on Scientific Projects Database; Copper; Environmental Risk Factor; Fluorescence; Funding; Grant; In Vitro; Institution; Ions; Laboratories; Ligands; Link; Metals; N-terminal; Neurodegenerative Disorders; Oxidation-Reduction; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathology; Proteins; Research; Research Personnel; Resources; Role; Source; Structure; United States National Institutes of Health; Work; alpha synuclein; base; insight; metal metabolism; mutant; protein complex

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. alpha-Synuclein (alpha-syn) has been implicated in the pathogenesis of a number of neurodegenerative diseases, the best known of which is Parkinson?s disease (PD). The sporadic form of PD has been linked to environmental factors that promote oxidative stress and aberrant redox-active metal metabolism, and metal-enhanced oligomerization of alpha-syn has been observed in vitro. Recent work in our laboratory has focused on the understanding the role of copper ions in the aggregation of alpha-syn. Our initial studies have shown that Cu(II) ions accelerate aggregation and that the copper ions are incorporated into alpha-syn, with the tightest affinity binding near the N-terminus (based on ICP-MS and fluorescence quenching). We propose Cu K-edge and EXAFS studies to examine the local structure of the Cu binding site in the WT protein and in related mutants. We will compliment these studies with XAS on synthetic N-terminal fragments in order to identify the Cu binding site ligands. Our studies will examine both Cu(II) and Cu(I) binding to the protein, as these results could potentially provide insights into the role of redox chemistry in alpha-syn pathology. Once the soluble Cu-protein complexes have been characterized, we are poised to examine alpha-syn fibrils generated in the presence of copper ions.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 α-突触核蛋白 (alpha-syn) 与许多神经退行性疾病的发病机制有关，其中最著名的是帕金森病 (PD)。散发性 PD 与促进氧化应激和异常氧化还原活性金属代谢的环境因素有关，并且在体外观察到金属增强的 α-syn 寡聚化。我们实验室最近的工作重点是了解铜离子在 α-syn 聚集中的作用。我们的初步研究表明，Cu(II) 离子会加速聚集，并且铜离子会并入 α-syn，在 N 末端附近具有最紧密的亲和力结合（基于 ICP-MS 和荧光猝灭）。我们提出 Cu K-edge 和 EXAFS 研究来检查 WT 蛋白和相关突变体中 Cu 结合位点的局部结构。我们将利用 XAS 对合成 N 末端片段进行这些研究，以鉴定 Cu 结合位点配体。我们的研究将检查 Cu(II) 和 Cu(I) 与蛋白质的结合，因为这些结果可能会深入了解氧化还原化学在 α-syn 病理学中的作用。一旦可溶性铜蛋白复合物得到表征，我们就准备检查铜离子存在下产生的 α-syn 原纤维。