Divergence of Carboxylate-Bridged Diiron Enzymes for Natural Product Biosynthesis

天然产物生物合成中羧酸桥二铁酶的分歧

基本信息

批准号：
10283095
负责人：
Thomas M Makris
金额：
$ 30.17万
依托单位：
NORTH CAROLINA STATE UNIVERSITY RALEIGH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-20 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10283095
关键词：
Acyl Carrier Protein Address Affect Alkane 1-monooxygenase Alkenes Anabolism Antibiotics Architecture Biochemical Biochemical Reaction Biology Carbon Chemicals Chemistry Complex Critical Pathways Cytochrome P450 Cytochrome a Decarboxylation Dioxygen Enzymes Evolution Exhibits Fatty Acids Folic Acid Funding Generations Genetic Geometry Grant Halogens Heme Hemerythrin Histidine Hydrogen Hydroxylation Iron Kinetics Laboratories Lead Ligands Ligation Link Membrane Metabolism Methane hydroxylase Molecular Mononuclear Natural Products Outcome Oxides Oxygen Pathogenicity Pathway interactions Proteins Reaction Role Scaffolding Protein Side Site Structure Work carboxylate catalyst chemical reaction cofactor desaturase flexibility halogenation interest lens metalloenzyme microbial oxidation pathogenic bacteria pharmacophore programs protein structure scaffold tool

项目摘要

Dinuclear iron enzymes (DIs) utilize a carboxylate- and histidine-coordinated cofactor to affect synthetically challenging biochemical reactions. The recognized roles for DIs have recently expanded to include several important natural product biosynthetic pathways, including the generation of folate in pathogenic bacteria, the modulation of antibiotic potency via halogen installation, and the synthesis of essential secondary metabolites through carbon-carbon bond scission. To understand the molecular basis for how a very similar cofactor and structural core can perform such diverse functions, we propose to study three newly discovered DIs that contain nearly identical coordination motifs and overall structural scaffolds, but orchestrate chemistry in ways that fundamentally differ from both each other and from well-studied DIs that instead perform the oxygenation of substrates. The proposed work will utilize an array of spectroscopic, kinetic, and genetic tools to identify key sites of the protein, substrate, and cofactor that enable such disparate activities. An elucidation of the structural basis for DI functional reprogramming will provide a biochemical template for the synthesis of new pharmacophores and the molecular basis for pathways that are critical for microbial proliferation and pathogenicity.

双核铁酶（DIS）利用羧酸盐和组氨酸协调的辅因子来影响合成具有挑战性的生化反应。最近公认的角色最近有扩展到包括几种重要的天然产物生物合成途径，包括致病细菌的叶酸产生，通过卤素调节抗生素效力安装，以及通过碳碳键合成必需的次级代谢物分裂。了解非常相似的辅因子和结构核的分子基础可以执行如此多样化的功能，我们建议研究包含的三个新发现的DIS 几乎相同的协调基序和整体结构支架，但在从根本上彼此不同的方式以及与经过充分研究的方式相反执行底物的氧合。拟议的工作将利用一系列光谱学，动力学和遗传工具，以识别蛋白质，底物和辅因子的关键位点这种不同的活动。阐明DI功能重编程的结构基础将提供一个生化模板，用于合成新的药算术和分子对于微生物增殖和致病性至关重要的途径的基础。