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Electron Transfer in the Cytochrome c - Cytochrome c Peroxidase Complex

细胞色素 c 中的电子转移 - 细胞色素 c 过氧化物酶复合物

基本信息

批准号：
8716459
负责人：
James Erman
金额：
$ 24.83万
依托单位：
Northern Illinois University
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
1988
资助国家：
美国
起止时间：
1988-04-01 至 1991-09-30
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=8716459&HistoricalAwards=false
关键词：
Electron Transfer Cytochrome Peroxidase Complex

项目摘要

The cytochrome c - cytochrome c peroxidase system provides an outstanding opportunity to uncover the intimate details of the structural requirements for peroxidase activity, as well as those for long range electron transfer between physiologically related heme proteins. The objective of this research proposal is to elucidate the detailed mechanism of cytochrome c peroxidase catalysis with special emphasis on characterizing the interaction between cytochrome c and cytochrome c peroxidase. Experiments designed to clarify the stoichiometry of binding between cytochrome c and cytochrome c peroxidase will be performed to determine if, and under what conditions, more than one cytochrome c may bind to cytochrome c peroxidase. The specificity of binding at the high affinity binding site will be explored by utilizing a variety of cytochrome cs isolated from different sources and through site-directed mutagenesis of cytochrome c peroxidase. Transient state kinetic techniques will be used to investigate the rate of electron transfer between ferrocytochrome c and the oxidized intermediates of cytochrome c preoxidase. The specificity of the reaction dynamics will be evaluated by using cytochrome c from various sources and the role of specific amino acid residues in the enzyme will be explored by site-directed mutagenesis.

细胞色素c -细胞色素c过氧化物酶系统提供了一种这是一个绝佳的机会来揭露过氧化物酶活性的结构要求，与那些用于长距离电子在生理学相关的血红素蛋白。的目的研究建议是阐明详细的机制，细胞色素c过氧化物酶催化，特别强调表征细胞色素c和细胞色素c过氧化物酶实验旨在阐明细胞色素c和细胞色素c之间结合的化学计量 c过氧化物酶将进行，以确定是否，什么样的条件下，一个以上的细胞色素c可以结合到细胞色素c过氧化物酶结合的特异性，高亲和力结合位点将通过利用从不同来源分离的各种细胞色素c，通过细胞色素c过氧化物酶的定点突变。瞬态动力学技术将用于研究电子转移的速率，铁细胞色素c和氧化的中间体细胞色素c前氧化酶反应的特异性动力学将通过使用来自各种细胞色素c进行评价。来源和特定氨基酸残基的作用，将通过定点诱变来探索酶。