Studies of environmentally relevant molybdenum enzymes

环境相关钼酶的研究

基本信息

批准号：
7491594
负责人：
Russ Hille
金额：
$ 28.2万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
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/7491594
关键词：
Active Sites Amino Acids Arsenates Arsenic Arsenites Behavior Biological Models Catabolism Catalysis Chemicals Chemistry Climate Depth Dimethyl Sulfoxide Drug Metabolic Detoxication Electronics Environment Enzymatic Biochemistry Enzymes Eukaryota Eukaryotic Cell Family Goals Hand Health Human Inorganic Sulfates Kinetics Label Metabolic Biotransformation Metals Methodology Molybdenum Nature Oxidases Oxygen Plants Play Process Property Protocols documentation Raman Spectrum Analysis Range Reaction Research Personnel Role Site Site-Directed Mutagenesis Sleep Initiation and Maintenance Disorders Source Structure Sulfites Sulfur Unspecified or Sulfate Ion Sulfates Variant Vertebrates Work arsenite oxidase base circular magnetic dichroism computer studies density design dimethyl sulfoxide reductase enzyme structure greenhouse gases in vivo member microorganism mutant oxidation prevent programs theories

项目摘要

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 constrasting 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 targetting specific active site amino acid residues will also be examined to evaluate their roles in catalysis.

拟议的工作重点是三种与环境相关的含钼酶：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 还原为抗温室气体 DIMS，因此不仅在全球硫循环中发挥着重要作用，而且在还可以调节气候。第二种酶催化亚砷酸盐氧化成砷酸盐，砷在环境中生物转化的重要一步，代表着解毒发现它的微生物的机制。它是同一个家族的成员含钼酶作为 DMSO 还原酶，但具有代表 DMSO 还原酶中观察到的变化。来自高等真核生物（脊椎动物和植物）催化硫分解代谢的最后一步，即亚硫酸盐氧化为硫酸盐，并防止高活性亚硫酸盐在体内的有害积累。拟议工作的总体目标是更全面地了解这些酶在其作用背景下的作用机制结构，比较和对比他们的行为。该方法背后的指导假设是酶的功能和催化能力由活性位点的物理和电子结构决定。具体目标包括快速动力学研究以及光谱和计算工作确定酶活性位点的电子结构。对于 DMSO 还原酶和亚硫酸盐氧化酶，针对特定活性位点氨基酸残基的定点突变体也将被检查以评估它们在催化中的作用。