BIOLOGICAL OXIDATION MECHANICS

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。研究方法： 方法将是研究具有代表性的特定酶 黄素蛋白的各种类别和模型系统的研究。 将在相当大程度上利用稳态动力学与 快速反应动力学研究和置换技术 黄素蛋白与合成黄素的天然辅酶，在 以测试可能的机制，并探索 周围环境中紧邻蛋白质结合的黄素。这个 后者在提供有关 蛋白质中结构不可用的活性部位区域 来自X射线结晶学。它还提供了一种强大的方式 在这种情况下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 L乳酸氧化酶基因的克隆及其活性测定 蛋白质的氨基酸序列，以帮助结晶学 确定其三维结构，并使用现场- 定向诱变在酶机制研究中的应用。

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.