CHAPERONE PROTEINS FOR COPPER/ZINC SUPEROXIDE DISMUTASE

铜/锌超氧化物歧化酶的伴侣蛋白

• 批准号: 2640278
• 负责人: TRACEY D RAE
• 金额: $ 2.62万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2640278
• 关键词: copper; metalloenzyme; molecular chaperones; nuclear magnetic resonance spectroscopy; protein engineering; protein structure function; recombinant proteins; superoxide dismutase; zinc

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.

根据最近的许多理论， 在各种疾病状态（ALS，关节炎， 缺血/再灌注损伤，癌症），阐明机制， 所述细胞管理已知产生氧化的催化金属 损害已成为一个关键问题。 研究中描述的 拟议的项目审查了两个潜在的铜输送功能 细胞溶质蛋白，HAH 1和CCS 1。假设这两个人 蛋白质将铜离子从膜转运蛋白转运到 抗氧化酶Cu、Zn超氧化物歧化酶（SOD）。 表达 E.大肠杆菌细胞将提供足够的数量， 结构表征以及金属结合的评价 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 绝佳的价钱 这些蛋白对铜插入apo-SOD的促进作用 将通过SOD活性测定（细胞色素c，NBT）进行评价， 铜转移事件。其他蛋白质作为金属的补充 如果HAH 1和CCS 1不直接促进， 铜插入。