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

• 批准号: 10157289
• 负责人: Amanda Byer
• 金额: $ 6.64万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 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/10157289
• 关键词: ATP phosphohydrolase; Active Sites; Address; Affect; Allosteric Regulation; Anabolism; Binding Sites; Biochemical; Biochemical Pathway; Biochemistry; Bioinformatics; Biological Models; Biological Process; Biophysics; Catalysis; Chemistry; Chlorophyll; Complex; Coupled; Coupling; Cryoelectron Microscopy; 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; Postdoctoral Fellow; Property; Protein Conformation; Protein Subunits; Proteins; Reaction; Regulation; Research; Roentgen Rays; Role; S-Adenosylmethionine; Sampling; Shapes; Side; Site; Source; Structure; Sulfur; Techniques; Testing; Tetrapyrroles; Therapeutic; Training; Transition Elements; United States National Institutes of Health; Universities; Variant; active control; chemical reaction; design; dimer; evidence base; improved; interfacial; metalloenzyme; oxidation; pi bond; prevent; response; single bond; structural biology; technique development; tool

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.

项目概要/摘要 大自然利用蛋白质变构来控制化学反应中的长程电子流 对于生命来说，金属和金属团簇通常是促进电子转移的关键成分。 已知用于门控金属酶中电子流的变构方法是交替位点反应性， 因此，根据构象，反应活性从酶的一半切换到另一半 氧化还原酶具有核心α2β2异四聚体结构，是铁硫簇。 采用交替位点反应性的金属酶，可以作为宝贵的模型 系统定义金属酶变构所需的结构基序和机制 符合 NIH NIGMS 药理学、生理学和生化化学的规定。 “提高对基本生物过程的分子水平理解和 发现它们的控制方法”，我们建议研究两种这样的氧化还原酶， 固氮酶样暗操作脱植基叶绿素氧化还原酶 (DPOR) 和叶绿素 氧化还原酶（COR），以确定长程相关运动如何促成交替- 我们将使用电子显微镜和小角度工具进行位点反应。 X 射线散射技术检查具有不对称构象的酶中间体 构象。