CHAPERONE PROTEINS FOR COPPER/ZINC SUPEROXIDE DISMUTASE

鉴于最近有许多关于 各种疾病(肌萎缩侧索硬化症、关节炎、 缺血/再灌注损伤，癌症)，阐明其机制 哪些细胞管理已知的产生氧化的催化金属 损害已成为一个关键问题。这项研究描述在 拟议的项目审查了两个潜在的铜输送功能 胞浆蛋白、HAH1和CCS1。假设是这两个人 蛋白质将铜离子从膜转运蛋白转运到 抗氧化酶铜锌超氧化物歧化酶(SOD)。的表达 大肠杆菌细胞中的蛋白质将提供足够的量来 金属结合的结构表征及评价 亲和力和金属转移化学。汞-199核磁共振和X射线 吸收将用于探测金属结合的结构 地点。这些蛋白质促进铜插入载脂蛋白超氧化物歧化酶 将通过测定(细胞色素c，NBT)的超氧化物歧化酶活性在 铜转移事件。在金属中作为补体的其他蛋白质 如果HAH1和CCS1不直接促进转移，将寻求转移 铜插入物。

In light of the many recent theories regarding the involvement of reactive oxygen species in various disease states (ALS, arthritis, ischemia/reperfusion damage, cancer), elucidation of the mechanisms by which cells manage the catalytic metals known to produce oxidative damage has become a critical issue. The research described in the proposed project examines the potential copper-delivery function of two cytosolic proteins, HAH1 and CCS1. The hypothesis is that these two proteins transport copper ions from the membrane transport protein to the antioxidant enzyme Cu,Zn superoxide dismutase (SOD). Expression of the proteins in E. coli cells will provide sufficient quantities for structural characterization as well as evaluation of metal binding affinities and metal transfer chemistry. Mercury-199 NMR and X-ray absorption will be used to probe the structure of the metal binding site. Facilitation of copper insertion into apo-SOD by these proteins will be evaluated by assays (cytochrome c, NBT) of SOD activity during the copper-transfer event. Other proteins acting as complements in metal transfer will be sought if HAH1 and CCS1 do not directly facilitate copper insertion.