Spectroscopic Investigations of Metalloenzyme Mechanisms

金属酶机制的光谱研究

• 批准号: 10552244
• 负责人: R David Britt
• 金额: $ 43.87万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552244
• 关键词: Active Sites; Anabolism; Antibiotics; Area; Binding; Binding Proteins; Biochemical; Biochemistry; Catalysis; Catalytic DNA; Cells; Collaborations; Complex; Dangerousness; Disease; Electron Spin Resonance Spectroscopy; Electrons; Enzymes; Frequencies; Goals; Health; Heart; Homeostasis; Human; Investigation; Ions; Knowledge; Laboratories; Lanthanoid Series Elements; Metals; Methods; Nature; Nitrogenase; Nuclear; Oxygen; Peptides; Play; Protein Engineering; Proteins; Protons; Reaction; Regulation; Ribosomes; Role; Site; Spin Labels; Techniques; Water; Work; circadian pacemaker; experimental study; fluopsin; iron hydrogenase; metalloenzyme; oxidation; photosystem; tool

Enzymes using metal centers and/or organic radicals play many crucial roles in the fundamental biochemistry of human health, with deficiencies in their bioassembly or enzymatic functions associated with various diseases. The R. David Britt laboratory is using advanced spectroscopic techniques, such as multifrequency electron paramagnetic resonance (EPR), to understand the assembly and catalytic mechanism of a number of such metal and radical centers. Many important enzymes involved in multielectron oxidation or reduction reactions employ metal clusters in their catalysis. The Britt laboratory is studying how such clusters are assembled by identifying and interrogating assembly intermediates with their spectroscopic methods. For example, the [Fe-Fe] hydrogenase enzyme uses a complex multinuclear Fe-S “H-cluster”, containing organometallic Fe-CO and Fe-CN components, to catalyze reversible interconversion of H2 with protons and electrons. How does nature safely assemble such a center involving potentially dangerous CO and CN- species? Other experiments are unraveling the biosynthesis of the complex Fe-S “M-cluster” at the heart of the nitrogenase enzyme, which can incorporate Mo or V or an additional Fe in its active site. We are studying the biosynthesis of an interesting Cu(II) containing antibiotic, Fluopsin C. Radical S-adenosylmethione (rSAM) enzymes carry out many interesting reactions. In one interesting area, we are studying how they transform peptides into ribosomally-synthesized and post-translationally modified peptide (RiPP) products. Regulation of metal composition in cells is crucial, and we are examining a number of metal- binding proteins involved in metal ion sequestration and homeostasis, including a new project examining lanthanide binding in proteins such as lanmodulin. We are targeting a number of de novo designed proteins for detailed characterization, and we are starting new collaborations examining the reactivity of artificial metalloenzymes and DNAzymes. We continue to collaborate and provide advanced EPR support in a number of interesting metalloenzyme and radical enzyme arenas, including work to cryotrap, and characterize with high field EPR, the transient oxygen- generating S4 state of the photosystem II water oxidizing enzyme. And using site directed spin labeling as a tool, we are probing the dynamics of an all-protein biochemical oscillator that serves as nature’s simplest circadian clock.

使用金属中心和/或有机自由基的酶在基本原理中发挥着许多关键作用。 人类健康的生物化学，其生物组装或酶功能存在缺陷 与各种疾病有关。 R. David Britt 实验室正在使用先进的光谱仪 技术，例如多频电子顺磁共振（EPR），以了解 许多此类金属和自由基中心的组装和催化机制。许多 参与多电子氧化或还原反应的重要酶采用金属 在其催化作用下形成团簇。布里特实验室正在研究这些簇是如何组装的 用光谱方法识别和询问组装中间体。为了 例如，[Fe-Fe]氢化酶使用复杂的多核 Fe-S“H 簇”， 含有有机金属 Fe-CO 和 Fe-CN 组分，催化可逆相互转化 H2 与质子和电子。大自然如何安全地组建这样一个中心？ 有潜在危险的 CO 和 CN- 物质吗？其他实验正在解开生物合成的谜团 位于固氮酶核心的复杂 Fe-S“M 簇”，它可以结合 Mo 或 V 或在其活性位点添加 Fe。我们正在研究一种有趣的生物合成 含有抗生素 Fluopsin C 的 Cu(II)。自由基 S-腺苷甲硫酮 (rSAM) 酶进行 许多有趣的反应。在一个有趣的领域，我们正在研究它们如何转化肽 转化为核糖体合成和翻译后修饰肽 (RiPP) 产品。 细胞中金属成分的调节至关重要，我们正在研究许多金属成分 参与金属离子螯合和稳态的结合蛋白，包括一个新项目 检查兰莫杜林等蛋白质中的镧系元素结合。我们的目标是一些德 novo 设计了用于详细表征的蛋白质，我们正在开始新的合作 检查人工金属酶和脱氧核糖核酸酶的反应性。我们继续合作 并在一些有趣的金属酶和自由基酶方面提供先进的EPR支持 领域，包括冷冻阱工作，并用高场 EPR 表征，瞬态氧- 产生光系统II水氧化酶的S4状态。并使用站点定向旋转 标记作为一种工具，我们正在探索全蛋白质生化振荡器的动力学，该振荡器服务于 作为自然界最简单的生物钟。

项目成果

Structural Properties and Catalytic Implications of the SPASM Domain Iron-Sulfur Clusters in Methylorubrum extorquens PqqE.

• DOI: 10.1021/jacs.0c02044
• 发表时间: 2020-07
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Wen Zhu;Lindsey M. Walker;Lizhi Tao;A. Iavarone;Xuetong Wei;R. Britt;S. Elliott;J. Klinman;J. Klinman

Elucidation of a Copper Binding Site in Proinsulin C-peptide and Its Implications for Metal-Modulated Activity.

• DOI: 10.1021/acs.inorgchem.0c01212
• 发表时间: 2020-07-06
• 期刊: Inorganic chemistry
• 影响因子: 4.6
• 作者: Stevenson MJ;Janisse SE;Tao L;Neil RL;Pham QD;Britt RD;Heffern MC
• 通讯作者: Heffern MC

Structure and Reactivity of a High-Spin, Nonheme Iron(III)- Superoxo Complex Supported by Phosphinimide Ligands.

磷酰亚胺配体支持的高自旋非血红素铁 (III)-超氧复合物的结构和反应性。

• DOI: 10.1021/jacs.1c05276
• 发表时间: 2021
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Winslow,Charles;Lee,HeuiBeom;Field,MackenzieJ;Teat,SimonJ;Rittle,Jonathan
• 通讯作者: Rittle,Jonathan

Chemical structure and bonding in a thorium(iii)-aluminum heterobimetallic complex.

• DOI: 10.1039/c8sc01260a
• 发表时间: 2018-05-14
• 期刊: Chemical science
• 影响因子: 8.4
• 作者: Altman AB;Brown AC;Rao G;Lohrey TD;Britt RD;Maron L;Minasian SG;Shuh DK;Arnold J
• 通讯作者: Arnold J

Proposed Mechanism for the Biosynthesis of the [FeFe] Hydrogenase H-Cluster: Central Roles for the Radical SAM Enzymes HydG and HydE.

• DOI: 10.1021/acsbiomedchemau.1c00035
• 发表时间: 2022-02-16
• 期刊: ACS bio & med chem Au
• 作者: Britt RD;Tao L;Rao G;Chen N;Wang LP
• 通讯作者: Wang LP