STUDIES OF TRIMETHYLAMINE DEHYDROGENASE

三甲胺脱氢酶的研究

基本信息

批准号：
6525483
负责人：
Russ Hille
金额：
$ 16.5万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-09-01 至 2004-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6525483
关键词：
X ray crystallography aminoacid electron spin resonance spectroscopy electron transport enzyme activity enzyme mechanism flavins intermolecular interaction metalloenzyme methylation oxidizing agents oxidoreductase protein structure function site directed mutagenesis tertiary amine

项目摘要

DESCRIPTION: (adapted from applicant's abstract) The overall goal of the proposed research is to gain deeper insight into the mechanism of action of trimethylamine dehydrogenase (TMADH), a member of an important group of enzymes that contain multiple redox-active centers and which have overall catalytic mechanisms involving electron transfer between these sites. In addition to their physiological significance, these proteins serve as extremely useful systems in which to examine the factors which govern the magnetic and electron-transfer interactions between such centers. The reaction catalyzed by TMADH, the oxidative demethylation of trimethylamine to dimethylamine and formaldehyde, is of considerable intrinsic interest in understanding the chemistry available to the isoalloxazine ring in biological systems. This chemistry is relevant to a number of other enzymes, including such clinically important enzymes as monoamine oxidase. The proposed work builds on previous work by the PI and has the following principal objectives: Examination of the role of specific amino acid residues in the reaction catalyzed by TMADH; characterization of the reaction of TMADH using time-resolved crystallography; characterization of the unusually strong magnetic interaction between the flavin and Fe4S4 semiquinone centers of TMADH; examination of the electrochemical and electron-transferring properties of TMADH; and characterization of the interaction between trimethylamine dehydrogenase and its physiological electron acceptor, and electron-transferring flavoprotein. Guided by the three-dimensional structure of trimethylamine dehydrogenase, the roles of specific amino acid residues in the active site of the enzyme with regard to each of these areas will be addressed by site-directed mutagenesis. The intention of this work is to construct a comprehensive picture of the relationship of structure to function in trimethylamine dehydrogenase that furthers our basic understanding of the mechanism of this and related enzymes.

描述：（改编自申请人的摘要）拟议的研究是为了深入了解三甲胺脱氢酶（TMADH），一组重要酶的成员其中包含多个氧化还原活性中心，并且具有总体催化这些位点之间涉及电子转移的机制。此外它们的生理意义，这些蛋白质非常有用检查控制磁性因素的系统这些中心之间的电子转移相互作用。反应由 tmadh，三甲胺对二甲胺和二甲胺的氧化脱甲基化和甲醛，在理解在生物系统中可用于异丙沙嗪环的化学。这化学与许多其他酶有关，包括这种临床重要酶作为单胺氧化酶。拟议的工作是基于PI以前的工作，并具有以下内容主要目标：检查特定氨基酸残基的作用在tmadh催化的反应中； tmadh反应的表征使用时间分辨的晶体学；异常强壮的表征黄素和Fe4s4 semiquinone中心之间的磁相互作用；检查电化学和电子转移特性 tmadh;和三甲胺之间的相互作用的表征脱氢酶及其生理电子受体，以及电子转移黄蛋蛋白。在三维结构的指导下三甲胺脱氢酶，特异性氨基酸残基在关于这些区域的每个区域的酶的活跃部位将是通过定向诱变解决。这项工作的目的是构建结构与功能关系的全面图片在三甲胺脱氢酶中，我们对该酶的机制和相关酶。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Spectroscopic and functional properties of novel 2[4Fe-4S] cluster-containing ferredoxins from the green sulfur bacterium Chlorobium tepidum.

来自绿色硫细菌温热绿菌的新型 2[4Fe-4S] 簇铁氧还蛋白的光谱和功能特性。

DOI：
10.1074/jbc.m107852200
发表时间：
2001
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Yoon,KS;Bobst,C;Hemann,CF;Hille,R;Tabita,FR
通讯作者：
Tabita,FR

A mechanism for substrate-Induced formation of 6-hydroxyflavin mononucleotide catalyzed by C30A trimethylamine dehydrogenase.

C30A 三甲胺脱氢酶催化底物诱导形成 6-羟基黄素单核苷酸的机制。

DOI：
10.1016/j.bmcl.2003.07.032
发表时间：
2003
期刊：
Bioorganic & medicinal chemistry letters
影响因子：
2.7
作者：
Lu,Xingliang;Nikolic,Dejan;Mitchell,DeannaJ;vanBreemen,RichardB;Mersfelder,JohnA;Hille,Russ;Silverman,RichardB
通讯作者：
Silverman,RichardB

Inactivation of C30A trimethylamine dehydrogenase by N-cyclopropyl-alpha-methylbenzylamine, 1-phenylcyclopropylamine, and phenylhydrazine.

N-环丙基-α-甲基苄胺、1-苯基环丙胺和苯肼使 C30A 三甲胺脱氢酶失活。