Radical SAM Enzymes: Molecular Mechanisms of Radical Initiation

自由基 SAM 酶：自由基引发的分子机制

• 批准号: 10389995
• 负责人: Joan B Broderick
• 金额: $ 9.23万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10389995
• 关键词: Administrative Supplement; Antibiotics; Biological; Chemicals; Complex; DNA Repair; Detection; Disease; Electron Nuclear Double Resonance; Electron Spin Resonance Spectroscopy; Enzymes; Freezing; Goals; Human; Infection; Life; Metals; Microbe; Molecular; Pharmacologic Substance; Process; Property; Reaction; Research; Structure; X-Ray Crystallography; beneficial microorganism; catalyst; chemical reaction; cofactor; experimental study; member; pathogenic microbe; photolysis

Broderick, Joan B. 5R35GM131889-02 Administrative Supplement for Purchase of a Rapid Freeze Quench / Stopped Flow Abstract/Summary of Project Radical SAM is one of the largest enzyme superfamilies known, with its members catalyzing a remarkable diversity of reactions in all domains of life. Radical SAM enzymes are involved in the synthesis of essential cofactors and antibiotics, repair of DNA damage, and the assembly of complex biological metal clusters, among many other reactions. The presence of radical SAM enzymes in humans as well as in both beneficial and pathogenic microbes lends high significance to understanding their fundamental properties and mechanisms. The proposed research will develop a critical understanding of radical SAM mechanisms, in particular the radical initiation process common to all enzymes in this large and diverse superfamily. Research efforts will focus on trapping radical intermediates using rapid freeze-quench, cryoreduction, annealing, and photolysis approaches. These intermediates will then be characterized by using spectroscopic approaches such as electron paramagnetic resonance and electron-nuclear double resonance, and structural approaches such as X-ray crystallography. The proposed supplement will allow the purchase of needed stopped-flow mixing/ rapid-freeze-quench and UV-visible detection capability to carry out these experiments..

Broderick，Joan B. 5R35GM131889-02 购买快速冷冻 /停止流量的行政补充 摘要/项目摘要 激进的SAM是已知的最大酶超家族之一，其成员催化A 生命的所有领域中反应的显着多样性。自由基SAM酶参与合成 必需辅助因子和抗生素，DNA损伤的修复以及复杂生物金属的组装 集群，以及许多其他反应。人类以及两者中存在激进的SAM酶的存在 有益和致病性微生物对了解其基本特性具有很高的意义 和机制。拟议的研究将对激进的SAM机制产生批判性理解， 特别是在这个大而多样化的超家族中，所有酶共有的激进启动过程。 研究工作将着重于使用快速冻结，冷冻度，捕获自由基中间体 退火和光解方法。然后，这些中间体将通过使用 光谱方法（例如电子顺磁共振和电子核双重） 共振和结构方法，例如X射线晶体学。拟议的补充剂将允许 购买所需的停止流量混合/快速冻结 - Quench和紫外可见的检测能力 进行这些实验。