RESPIRATORY PROTEIN COMPLEXES

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)复合体，这一建议反映了前NIH的评论 研究部分，建议将重点放在核磁共振研究上 非共价CCP/细胞色素c复合体。在这项提案中，我们打算 重点是对一种非生理性CCP复合体的比较研究： 在CCP和马细胞色素c之间；以及一种生理类型的CCP 复合体：在CCP和其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 从我们过去三年的工作中脱颖而出 沿着物种特有的路线。我们最有趣的结果之一就是 这一次是游离和结合的铁细胞色素c的拆分。 Ccp/细胞色素c非共价配合物的核磁共振波谱研究。 我们建议用核磁共振波谱进一步表征这些化合物， 阐明了络合解离速率常数，并测定了络合解离速率常数 与天然药物形成络合物对细胞色素c-核磁共振波谱的影响 野生型CCP(一种铁的、高自旋的血红素蛋白)，以及 氧化中间体，CCP-I(化合物-I)和CCP-II(化合物-II)，以及 氰化物抑制的CCP(CcPCN)。核磁共振检测络合物的作用 CCP时将尝试结合使用CcPCN和 亚铁和铁细胞色素c.

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.