CATALYTIC ROLE OF NI F430 IN METHYL-COM REDUCTASE

NI F430 在甲基-COM 还原酶中的催化作用

• 批准号: 2608706
• 负责人: Donald F Becker
• 金额: $ 2.99万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-11-27 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2608706
• 关键词: catalyst; cofactor; enzyme activity; enzyme mechanism; methylation; microorganism metabolism; oxidoreductase

Methyl-CoM reductase (MCR) catalyzes the final key step in methanogenesis. As a result, the determination of the MCR molecular mechanism is critical to the understanding of the unique metabolic pathway of methanogenesis. The objective of this proposal is to determine the catalytic role of the MCR cofactor, Ni F43O, during MCR catalysis. The specific aims to achieve this are the following: (a) to generate a fully active and stable form of MCR; (b) to characterize the catalytically competent Ni redox states of F430 and the interaction of methyl-SCoM with F430 during MCR catalysis; (c) to determine the overall catalytic mechanism of MCR and any rate limiting steps. The stopped-flow kinetic studies will include rapid freeze-quench techniques along with a variety of spectroscopic methods such a Uv- visible, EPR, and resonance Raman. The insights gained from the completion of these goals will contribute significantly to the understanding of the NCR molecular mechanism. It should be noted that I have recently isolated an active form of NCR which will allow me to begin to characterize the catalytic role of F430 while continuing to investigate the activation of MCR.

甲基CoM还原酶（MCR）催化最后的关键步骤 甲烷生成 因此，MCR分子的测定 这一机制对于理解这种独特的代谢机制至关重要。 甲烷生成途径。 这项建议的目的是 确定MCR辅因子Ni F43 O在MCR过程中的催化作用 催化作用 实现这一目标的具体目标如下： MCR的完全活性和稳定形式;（B）表征 F430的催化活性Ni氧化还原态及其相互作用 MCR催化期间甲基-SCoM与F430的反应;（c）确定 MCR的总体催化机理和任何速率限制步骤。 的 停流动力学研究将包括快速冷冻淬火 技术沿着与各种光谱方法，如UV- 可见光、EPR和共振拉曼。 从《经济学人》中获得的见解 这些目标的实现将大大有助于 了解NCR分子机制。 应当注意 我最近分离出了一种活跃的NCR 我开始描述F430的催化作用， 继续研究MCR的激活。