METHANE MONOOXYGENASE STRUCTURE/FUNCTION

甲烷单加氧酶结构/功能

• 批准号: 6018736
• 负责人: JOHN D LIPSCOMB
• 金额: $ 27.07万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6018736
• 关键词: Methanobacteriaceae; Mossbauer spectrometry; Raman spectrometry; active sites; chemical kinetics; circular dichroism; cofactor; crosslink; crystallization; electron nuclear double resonance spectroscopy; electron spin resonance spectroscopy; enzyme complex; enzyme mechanism; enzyme reconstitution; enzyme structure; enzyme substrate; iron; metalloenzyme; methane; methane monooxygenase; microorganism metabolism; nuclear magnetic resonance spectroscopy; oxidation reduction reaction; oxygenases; protein purification

DESCRIPTION: The PI proposes to investigate the 3D structure, active site architecture, catalytic mechanism and mechanism of activation of soluble methane monooxygenase from the Type II methanotroph Methylosinus trichosporum OB3b. This enzyme catalyzes the first step in the oxidation of methane to CO2 by methanogenic bacteria. In this way, the atmospheric egress of nearly all the enormous quantity of biogenic methane (a potent greenhouse gas) generated by anaerobic bacteria is prevented. MMO also adventitiously catalyzes the oxidation of many other hydrocarbons leading to applications in synthesis and biodegradation. Work to date suggests that the reaction is catalyzed by a cofactor not found in other oxygenases, implying a new strategy for oxygen activation. The proposed studies build on the very significant progress that has been made in the previous funding period, and focus on: elucidation of the physical and electronic structure of dinuclear iron cluster of the active site as well as the mechanism of oxygen activation catalyzed by this center; spectroscopic and kinetic characterization of the several reaction intermediates that have been identified in the catalytic sequence; elucidation of the protein-protein interactions between the three polypeptides of the enzyme that modulate its catalytic activity.

描述：PI提议研究3D结构，主动位点 结构，催化机制和可溶性激活机制 来自II型甲烷营养甲基素的甲烷单加氧酶 Trichosporum ob3b。 这种酶催化了氧化的第一步 甲烷通过甲烷基细菌。 这样，大气 几乎所有大量生物甲烷的出口（有效的 防止厌氧菌产生的温室气）。 也是MMO 异议地催化许多其他碳氢化合物的氧化导致 在合成和生物降解中的应用。 迄今为止的工作表明 该反应是由其他氧合酶中未发现的辅因子催化的， 意味着一种新的氧气激活策略。 拟议的研究建立 关于以前的资金取得的非常重大的进展 时期，专注于：阐明物理和电子结构 活性部位的双核铁簇以及 该中心催化的氧气激活；光谱和动力学 表征几个反应中间体 在催化序列中鉴定；阐明蛋白质蛋白 调节酶的三个多肽之间的相互作用 催化活性。