BIOLOGICAL OXIDATION MECHANICS

生物氧化机制

• 批准号: 2168536
• 负责人: VINCENT MASSEY
• 金额: $ 46.57万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2168536
• 关键词: Escherichia coli; NAD(P)H dehydrogenase; X ray crystallography; aminoacid oxidase; charge transfer complex; chemical reaction; cofactor; electron transport; enzyme mechanism; enzyme model; enzyme structure; enzyme substrate complex; flavins; flavoproteins; fluorescence spectrometry; hydroxybenzoate; hydroxylation; lactate dehydrogenases; oxidation; oxidoreductase; oxygenases; protonation; radiotracer; spectrometry

The main aim of the research project is an understanding of the chemical reactivity of flavin coenzymes, and how this reactivity is used and controlled in biological systems. The methods of approach will be the study of specific enzymes representative of the various classes of flavoproteins and studies of model systems. Considerable use will be made of steady state kinetics coupled with rapid reactior kinetics studies, and of the technique of replacing the natural coenzymes of flavoproteins with synthetic flavins, in order to test possible mechanisms, and to probe the nature of the environment immediately around the protein-bound flavin. The latter is particularly important in providing information about the active site region in proteins where the structure is not available from X-ray crystallography. It also offers a powerful way for investigating dynamic aspects of protein structure in the case of enzymes where the crystal structure is known. We also plan to clone the gene for L-lactate oxidase in order to determine the amino acid sequence of the protein, to aid in the crystallographic determination of its three-dimensional structure, and to use site- directed mutagenesis in study of the enzyme mechanism.

研究项目的主要目的是了解 黄素辅酶的化学反应性，以及这种反应性如何 在生物系统中使用和控制。 方法 方法将是对代表的特定酶的研究 各种类黄酮蛋白和模型系统的研究。 大量使用将与稳态动力学相结合 快速反应动力学研究和更换的技术 黄素蛋白与合成黄素的天然辅酶在 为了测试可能的机制，并探究 立即在蛋白质结合黄素周围的环境。 这 后者在提供有关有关的信息时尤其重要 蛋白质中的活跃位点区域，该蛋白质不可用 来自X射线晶体学。 它还为 研究蛋白质结构的动态方面 已知晶体结构的酶。 我们还计划 克隆L乳酸氧化酶的基因，以确定 蛋白质的氨基酸序列，有助于晶体学 确定其三维结构，并使用站点 - 在研究酶机制的研究中定向诱变。