Mechanistic Studies of Type II IPP Isomerase

II型IPP异构酶的机理研究

• 批准号: 6789763
• 负责人: Steven C. Rothman
• 金额: $ 4.3万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6789763
• 关键词: NAD(H) phosphate; X ray crystallography; active sites; bacteria; bacterial proteins; chemical structure function; deuterium; enzyme activity; enzyme mechanism; flavin mononucleotide; gene mutation; isomerase; oxidoreductase; postdoctoral investigator; site directed mutagenesis; spectrometry; stereochemistry

DESCRIPTION (provided by applicant): A recent report describes the cloning and preliminary characterization of an unusual flavin mononucleotide (FMN) dependent enzyme, the type II isopentenyl diphosphate (IPP) isomerase. IPP isomerases interconvert isopentenyl diphosphate and dimethylallyl diphosphate, the metabolic building blocks for a wide array of biological isoprenoid compounds. The newly identified type II enzyme represents an attractive antimicrobial drug target as sequence analyses suggest that it is essential in the pathogen Staphylococcus aureus and is not present in humans. The initial study with the type II IPP isomerase indicated that catalysis requires both FMN and NADPH. The role is these two cofactors is unclear, given that isomerization does not entail a net oxidation/reduction and that the type l isomerase catalyzes a proton addition-proton elimination reaction that does not include a transient oxidation/reduction. This proposed work intends to elucidate the functions of the two cofactors, the chemical mechanism, and the roles of active site residues in the reaction catalyzed by type II isopentenyl diphosphate isomerase from Synechocystis sp. PCC 6803.

描述(由申请人提供)： 最近的一份报告描述了一种不常见的黄素单核苷酸(FMN)依赖酶--II型异戊烯基二磷酸(IPP)异构酶的克隆和初步鉴定。IPP异构酶相互转化异戊烯基二磷酸和二甲基烯丙基二磷酸，这是一系列生物异戊二烯类化合物的代谢构件。新发现的II型酶是一个有吸引力的抗菌药物靶点，因为序列分析表明，它在病原体金黄色葡萄球菌中是必不可少的，在人类中不存在。对II型IPP异构酶的初步研究表明，催化既需要FMN，也需要NADPH。鉴于异构化不需要净氧化/还原，并且L异构酶催化不包括瞬时氧化/还原的质子加成-质子消除反应，这两个辅助因素的作用尚不清楚。本工作旨在阐明这两个辅因子的功能、化学机制以及活性中心残基在聚球藻II型异戊烯基二磷酸异构酶催化反应中的作用。PCC 6803。