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

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 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。

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 (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.