Structural Studies of Alternating-site Reactivity in Nitrogenase-like Oxidoreductases

固氮酶样氧化还原酶的交替位点反应性的结构研究

基本信息

批准号：
10592277
负责人：
Amanda Byer
金额：
$ 7.38万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-15 至 2024-04-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10592277
关键词：
ATP phosphohydrolase Active Sites Address Affect Allosteric Regulation Anabolism Binding Binding Sites Biochemical Biochemical Pathway Biochemistry Bioinformatics Biological Models Biological Process Biophysics Catalysis Chemistry Chlorophyll Complex Coupled Coupling Cryoelectron Microscopy Darkness Drug Targeting Electron Microscopy Electron Transport Electrons Enzymes Goals Hydrogenase Ions Iron Knowledge Length Life Metals Methods Mission Molecular Molecular Conformation Montana Motion Movement National Institute of General Medical Sciences Nature Nitrogenase Organism Outcome Oxidation-Reduction Oxidoreductase Oxygen Pharmaceutical Preparations Pharmacology Physiology Positioning Attribute Postdoctoral Fellow Proliferating Property Protein Conformation Protein Subunits Proteins Reaction Regulation Research Roentgen Rays Role S-Adenosylhomocysteine S-Adenosylmethionine Sampling Shapes Side Site Source Structure Sulfur Techniques Testing Tetrapyrroles Therapeutic Training Transition Elements United States National Institutes of Health Universities Variant chemical reaction design dimer evidence base improved interfacial metalloenzyme oxidation pi bond prevent response single bond structural biology technique development tool

项目摘要

Project Summary / Abstract Nature uses protein allostery to control long-range electron flow in chemical reactions essential for life. Metals and metal-clusters are often key components facilitating this electron transfer. A known allosteric method for gating electron flow in metalloenzymes is alternating-site reactivity, wherein reactivity switches from one half of the enzyme to the other upon conformational changes. Oxidoreductases, with a core α2β2 heterotetrameric structure, are iron-sulfur cluster metalloenzymes that employ alternating-site reactivity and can serve as an invaluable model system to define the structural motifs and mechanisms required for metalloenzyme allosteric regulation. In line with the NIH NIGMS Pharmacology, Physiology, and Biochemical Chemistry mission of “improving molecular level understanding of fundamental biological processes and discovering approaches to their control”, we propose to study two such oxidoreductases, the nitrogenase-like dark-operative chlorophyllide oxidoreductase (DPOR) and chlorophyllide oxidoreductase (COR), to identify how long-ranged correlated motions contribute to alternating- site reactivity using structural biology tools. We will employ electron microscopy and small-angle X-ray scattering techniques to examine enzyme conformational intermediates with asymmetrical conformations.

项目摘要 /摘要大自然使用蛋白质变构来控制化学反应中的远程电子流。生命。金属和金属簇通常是支持该电子传输的关键组件。一个已知的变构方法用于金属酶中的门控电子流是改变位点反应性的，其中，反应性在构象时从酶的一半转换为另一个酶更改。氧化酶，具有核心α2β2杂元结构，是铁硫簇雇员交替反应性的金属酶可以作为宝贵的模型定义金属酶变构所需的结构基序和机制的系统规定。与NIH NIGMS药理学，生理学和生化化学一致 “提高分子水平了解基本生物学过程的使命和发现他们控制的方法”，我们建议研究两个这样的氧化还原酶，即氮酶样暗术叶绿素氧化还原酶（DPOR）和叶绿素叶绿素氧化还原酶（COR），以确定长期相关运动有助于替代运动使用结构生物学工具的站点反应性。我们将采用电子显微镜和小角度 X射线散射技术检查酶会议中间体与不对称的中间体构象。