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.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 α-突触核蛋白（α-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。s病（PD）。散在形式的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，并且已经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集中在理解铜离子在α-syn聚集中的作用。我们的初步研究表明，Cu（II）离子加速聚集，并且铜离子被掺入α-syn中，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）。我们建议铜K-边缘和EXAFS研究，以检查本地结构的铜结合位点的WT蛋白和相关的突变体。我们将补充这些研究与XAS合成的N-末端片段，以确定铜结合位点配体。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为氧化还原化学在alpha-syn病理学中的作用提供见解。一旦可溶性铜蛋白复合物的特点，我们准备检查在铜离子的存在下产生的α-syn原纤维。