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个出色的机会，可以揭示过氧化物酶活性的结构要求，以及在生理相关的血红素蛋白之间远程电子转移的结构需求。这项研究建议的目的是阐明细胞色素C过氧化物酶催化的详细机制，特别强调了细胞色素C和细胞色素C过氧化物酶之间的相互作用。旨在阐明细胞色素C和细胞色素C过氧化物酶之间结合的化学计量的实验将进行，以确定是否以及在什么条件下，一个以上的细胞色素C可能与细胞色素C过氧化物酶结合。通过利用从不同来源分离的各种细胞色素Cs和通过位于细胞色素C过氧化物酶的位置诱变，将探索在高亲和力结合位点上结合的特异性。瞬态状态动力学技术将用于研究亚凝胶体C与细胞色素C前氧化酶的氧化中间体之间的电子转移速率。将通过使用来自各种来源的细胞色素C来评估反应动力学的特异性，特异性氨基酸残基在酶中的作用将通过定位的诱变来探索。