XAS STUDIES OF HEME-COPPER ASSEMBLY PROTEINS

血红素-铜组装蛋白的 XAS 研究

• 批准号: 7721908
• 负责人: Ninian J Blackburn
• 金额: $ 0.36万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721908
• 关键词: ATP Synthesis Pathway; Area; Bioenergetics; Biological Assay; Complement; Computer Retrieval of Information on Scientific Projects Database; Copper; Coupled; Data; Defect; Electron Transport; Funding; Genetic; Grant; Health; Heme; Homologous Gene; Human; Institution; Link; Lung diseases; Mammals; Metals; Mutation; Organism; Oxidases; Oxidation-Reduction; Oxygen; Pathway interactions; Play; Proteins; Research; Research Personnel; Resources; Role; Selenomethionine; Site; Source; Structure; United States National Institutes of Health; copper oxidase; cytochrome c oxidase; frontier; in vivo; programs

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. Heme-copper oxidases are the terminal oxidases of the electron transport chains of mammals and a wide variety of eukaryotic and prokaryotic organisms. They play a pivotal role in cellular bioenergetics and human health by converting the redox energy of oxygen reduction into a protonmotive force which is ultimately used to drive the endogonic synthesis of ATP. As the structure and catalytic mechanism have become better understood, the pathways leading to the assembly and metallation of the critical redox centers are emerging as the next frontier. This proposal aims will focus on the proteins involved in the assembly and metallation of the CuA center. Our experimental program is divided into two areas. (i) We will probe the function of the Sco homologue of B. subtilis via XAS structural studies coupled to in vivo functional assays of native proteins and Cu(I) and Cu(II)-site specific mutations; (ii) We will probe the mechanism of metal transfer between Sco and its partner protein, the CuA-containing CCO subunit II, using selenomethionine substitution to provide a site specific XAS probe of metal transfer reactivity. These studies will provide much-needed biophysical data on the proteins involved in cytochrome oxidase assembly, and will complement the extensive genetic evidence linking defects in CCO assembly with respiratory disease.

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。 血红素铜氧化酶是哺乳动物和多种真核和原核生物电子传递链的末端氧化酶。它们通过将氧还原的氧化还原能转化为质子动力，最终用于驱动 ATP 的内源合成，在细胞生物能学和人类健康中发挥着关键作用。随着结构和催化机制得到更好的理解，导致关键氧化还原中心组装和金属化的途径正在成为下一个前沿领域。该提案的目标将重点关注参与 CuA 中心组装和金属化的蛋白质。我们的实验计划分为两个区域。 (i) 我们将通过 XAS 结构研究与天然蛋白质以及 Cu(I) 和 Cu(II) 位点特异性突变的体内功能测定相结合，探索枯草芽孢杆菌的 Sco 同源物的功能； (ii) 我们将利用硒代蛋氨酸取代来探究 Sco 与其伴侣蛋白（含 CuA 的 CCO 亚基 II）之间的金属转移机制，以提供金属转移反应性的位点特异性 XAS 探针。这些研究将提供有关细胞色素氧化酶组装所涉及的蛋白质的急需的生物物理数据，并将补充将 CCO 组装缺陷与呼吸系统疾病联系起来的广泛遗传证据。