Structural investigations, at atomic level, of three enzymes comprising steps three, four and five of the mevalonate-independant pathway for isoprenoid biosynthesis

在原子水平上对三种酶进行结构研究，包括类异戊二烯生物合成的甲羟戊酸独立途径的第三步、第四步和第五步

• 批准号: 5422756
• 负责人: Dr. Florence Pojer
• 金额: --
• 依托单位: Structural Biology Laboratory (aufgelöst)
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2003
• 资助国家: 德国
• 起止时间: 2002-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5422756?language=en
• 关键词: Structural; investigations; atomic; level; three

The objective of this proposal is to understand the structural and mechanistic features governing the biosynthesis of isoprenoids and their chemical precursors in bacteria. Isoprenoids are the most structurally diverse family of compounds found in nature. More than 23.000 isoprenoid molecules are know to date [1]. Many isoprenoids have biotechnological applications as drugs, flavours, pigments, perfumes or agrochemicals. In living beings, isoprenoids have important roles in processes as diverse electron transport, reproduction, growth regulation, signal transduction and defence. I propose to use protein x-ray crystallography in an integrated fashion with functional experiments including site-directed mutagenesis and steady state kinetic analysis to address the stereochemical and kinetic basis for isoprenoid biosynthesis through the so-called mevalonate-independent or methyl-D-erythritol (MEP) pathway at atomic resolution [2]. This research proposal will focus on the structural elucidation of substrates complexed with kinetically impaired mutants of three enzymes comprising steps three, four, and five of the MEP pathway for isoprenoid biosynthesis. Since the MEP pathway is absent in humans and has been shown to be essential in bacteria, its enzymes are excellent candidates for new drug targets, for the treatment of both bacterial and parasitic infections.

本提案的目的是了解细菌中类异戊二烯及其化学前体生物合成的结构和机制特征。类异戊二烯是自然界中发现的结构最多样化的化合物家族。迄今已知超过23，000种类异戊二烯分子[1]。许多类异戊二烯具有作为药物、调味剂、颜料、香料或农用化学品的生物技术应用。在生物体内，类异戊二烯在多种电子传递、生殖、生长调节、信号转导和防御等过程中发挥重要作用。我建议使用蛋白质X射线晶体学与功能实验，包括定点诱变和稳态动力学分析，以解决通过所谓的甲羟戊酸盐独立或甲基-D-异戊烯（MEP）途径在原子分辨率下的类异戊二烯生物合成的立体化学和动力学基础[2]。这项研究计划将集中在与三种酶的动力学受损突变体复合的底物的结构阐明上，这些酶包括类异戊二烯生物合成的MEP途径的第三、四和五步。由于MEP途径在人类中不存在，并且已被证明在细菌中是必不可少的，因此其酶是用于治疗细菌和寄生虫感染的新药物靶点的优秀候选者。