Studies of environmentally relevant molybdenum enzymes

环境相关钼酶的研究

• 批准号: 7117987
• 负责人: Russ Hille
• 金额: $ 31.2万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7117987
• 关键词: Arabidopsis; Raman spectrometry; active sites; chemical kinetics; circular magnetic dichroism; electron spin resonance spectroscopy; enzyme activity; enzyme mechanism; enzyme structure; iron sulfur protein; molybdenum; oxidoreductase; plant proteins; protein protein interaction; site directed mutagenesis

DESCRIPTION (provided by applicant): The proposed work focuses on three molybdenum-containing enzymes of environmental relevance: DMSO reductase, arsenite oxidase and sulfite oxidase. The first of these catalyzes the reduction of DMSO to the anti-greenhouse gas DIMS, and as such plays an important role not simply in the global sulfur cycle but in modulating climate as well. The second enzyme catalyzes the oxidation of arsenite to arsenate, an important step in the biotransformation of arsenic in the environment that represents a detoxification mechanism for those microorganisms in which it is found. It is a member of the same family of molybdenum-containing enzymes as DMSO reductase, but has an active site structure that represents a variation on that seen in DMSO reductase. Sulfite oxidase from higher eukaryotes (both vertebrates and plants) catalyzes the final step in sulfur catabolism, the oxidation of sulfite to sulfate, and prevents the deleterioius accumulation of the highly reactive sulfite in vivo. The overall goal of the proposed work is to gain a more complete understanding of the mechanism of action of these enzymes in the context of their structures, comparing and contrasting their behavior. The guiding hypothesis behind the approach is that enzyme function and catalytic power are dictated by the physical and electronic structure of the active site. The Specific Aims include rapid kinetic studies as well as spectroscopic and computational work aimed at determining the electronic structures of the enzyme active sites. In the cases of DMSO reductase and sulfite oxidase, site-directed mutants targeting specific active site amino acid residues will also be examined to evaluate their roles in catalysis.

描述（由申请人提供）：拟议的工作重点是三种与环境相关的含氘酶：DMSO还原酶、亚砷酸盐氧化酶和亚硫酸盐氧化酶。其中第一个催化DMSO还原为反温室气体DIMS，因此不仅在全球硫循环中发挥重要作用，而且在调节气候方面也发挥重要作用。第二种酶催化亚砷酸盐氧化为砷酸盐，这是环境中砷生物转化的重要步骤，代表了发现它的微生物的解毒机制。它与DMSO还原酶属于含砷酶家族，但其活性位点结构与DMSO还原酶不同。来自高等真核生物（脊椎动物和植物）的亚硫酸盐氧化酶催化硫催化的最后一步，将亚硫酸盐氧化成硫酸盐，并防止高活性亚硫酸盐在体内的累积。拟议工作的总体目标是在其结构的背景下更全面地了解这些酶的作用机制，比较和对比它们的行为。这种方法背后的指导假设是，酶的功能和催化能力是由活性位点的物理和电子结构决定的。具体目标包括快速动力学研究以及旨在确定酶活性位点的电子结构的光谱和计算工作。在DMSO还原酶和亚硫酸盐氧化酶的情况下，还将检查靶向特定活性位点氨基酸残基的定点突变体，以评估它们在催化中的作用。