ENGINEERING OF NOVEL SUBSTRATE OXIDATION IN HEME ENZYMES

血红素酶中新型底物氧化的工程

• 批准号: 2391801
• 负责人: RABI A MUSAH
• 金额: $ 2.86万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2391801
• 关键词: active sites; cytochrome c peroxidase; enzyme substrate complex; protein engineering; protein structure function; synthetic enzyme

The goal of this project is to utilize engineered forms of the enzyme cytochrome c peroxidase to demonstrate the feasibility of rational protein design towards the accomplishment of oxidative catalysis of specific organic reactions. The discovery that the active site Trp- 191 in the oxidized "ES" state of CCP is reversibly oxidized to a stable cation free radical has prompted an interest in exploiting the chemistry inherent at this site for the possible binding and/or oxidation of organic molecules. To this end, construction of the W191G mutant, in which the active site tryptophan was replaced with glycine, was recently reported, and the feasibility of small molecule binding to this site was clearly demonstrated. Part I of this proposal involves the systematic determination of the factors that control substrate specificity and binding within the cavity. Part II of this proposal involves an investigation of the oxidative chemistry of organic substrates, such as the aminothiazole anti-thyroid drugs, that are known to react with heme enzymes. Part III of this proposed study utilizes the information gleaned from Parts I and II of this proposal, and focuses on the feasibility of utilizing the W191G cavity mutant for the purposes of catalyzing general oxidation of organic substrates from within the cavity.

该项目的目标是利用酶的工程形式 细胞色素c过氧化物酶证明合理的可行性 蛋白质设计以实现氧化催化 特定的有机反应。发现活性位点Trp- CCP氧化“ES”态的191被可逆地氧化成 稳定的阳离子自由基引起了人们对开发 该位点固有的化学性质，用于可能的结合和/或 有机分子的氧化。为此，建设 W191G 突变体，其中活性位点色氨酸被替换为 最近报道了甘氨酸，以及小分子的可行性 清楚地证明了与该网站的绑定。本部分第一部分 提案涉及系统确定影响因素 控制腔内的底物特异性和结合。第二部分 该提案涉及氧化化学的研究 有机底物，例如氨基噻唑抗甲状腺药物， 已知与血红素酶发生反应。本提议的第三部分 研究利用了从本报告第一部分和第二部分收集的信息 提案，重点讨论利用W191G的可行性 空腔突变体用于催化一般氧化 腔内的有机基质。