CHARACTERIZATION OF RESPIRATORY PROTEIN COMPLEXES

呼吸蛋白复合物的表征

• 批准号: 3305480
• 负责人: James D. Satterlee
• 金额: $ 12.97万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1996-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3305480
• 关键词: covalent bond; cytochrome c; enzyme activity; enzyme complex; enzyme mechanism; iron compounds; isozymes; nuclear magnetic resonance spectroscopy; peroxidases; respiratory protein; yeasts

Using the results from our previous work on yeast cytochrome c peroxidase (CcP) complexes, this proposal reflects the comments of the previous NIH Study Section which recommended a proposal tightly focused on NMR studies of noncovalent CcP/cytochrome c complexes. In this proposal we intend to focus on comparative studies of a nonphysiological type of CcP complex: between CcP and horse cytochrome c; and a physiological type of CcP complex: between CcP and its natural redox partner yeast iso- 1 cytochrome c. This discrimination of complex type is based upon results from our work during the past three years, which segregate themselves along species-specific lines. One of our most interesting results during this time has been the resolution of free and bound ferricytochrome c resonances in NMR spectra of CcP/ferricytochrome c noncovalent complexes. We propose to further characterize these complexes by NMR spectroscopy, elucidate the complex dissociation rate constants and determine the effects on cytochrome c NMR spectra of forming complexes with native, wild-type CcP (a ferric, high-spin heme protein), and as well, with the oxidized intermediates, CcP-I (compound-I) and CcP-II (compound-II), and cyanide-inhibited CcP (CcPCN). NMR detection of effects of complex formation upon CcP will be attempted using CcPCN in combination with ferro- and ferricytochromes c.

利用我们以前对酵母细胞色素c过氧化物酶的研究结果， (CcP)复杂，该提案反映了前NIH的意见 研究部分，建议密切关注NMR研究 非共价CcP/细胞色素c复合物。 在本提案中，我们打算 专注于非生理类型的CcP复合物的比较研究： CcP和马细胞色素c之间的关系;以及CcP的生理类型 复合物：CcP和它的天然氧化还原伙伴酵母iso- 1之间 细胞色素c。这种复杂类型的区分是基于结果的 在过去的三年里，我们的工作， 沿着物种特异性线。 我们最有趣的结果之一， 这一次是游离和结合的铁细胞色素C的分辨率 CcP/铁细胞色素c非共价复合物的NMR光谱中的共振。 我们建议通过NMR光谱进一步表征这些配合物， 阐明复合物的解离速率常数，并确定 与天然的， 野生型CcP（一种铁，高自旋血红素蛋白），以及，与 氧化的中间体，CcP-I（化合物-I）和CcP-II（化合物-II），和 氰化物抑制的CcP（CcPCN）。 配合物效应的NMR检测 将尝试使用CcPCN结合 铁和铁细胞色素c.