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中起许多至关重要的作用 人类健康的生物化学，其生物组装或酶促功能缺陷 与各种疾病有关。 R. David Britt实验室正在使用高级光谱学 诸如多频电子顺磁共振（EPR）之类的技术，以了解 许多此类金属和激进中心的组装和催化机制。许多 参与多电体氧化或还原反应的重要酶员工金属 集群在催化中。布里特实验室正在研究如何通过 识别和审问组装中间体的光谱方法。为了 例如，[Fe-Fe]氢化酶使用复杂的多核Fe-S“ H-Cluster”， 包含有机Fe-CO和Fe-CN组件，以催化可逆互转换 带有质子和电子的H2。自然如何安全地组装这样的中心 潜在的危险CO和CN种类？其他实验正在阐明生物合成 氮酶心脏中心的复合fe-s“ m-cluster” MO或V或在其活跃部位的额外FE。我们正在研究一个有趣的生物合成 含有抗生素，荧光素C.自由基S-腺苷甲硫代（RSAM）酶进行的Cu（II）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改变肽 进入核糖体合成和翻译后修饰的肽（RIPP）产物。 细胞中金属成分的调节至关重要，我们正在检查许多金属 涉及金属离子隔离和稳态的结合蛋白，包括一个新项目 检查诸如lanmodulin之类的蛋白质中的灯笼结合。我们针对多个DE Novo设计了用于详细表征的蛋白质，我们正在开始新的合作 检查人工冶金和dnazymes的反应性。我们继续合作 并在许多有趣的金属酶和自由基酶中提供高级EPR支持 竞技场，包括冻结的工作，并以高场EPR为特征，瞬态氧气 生成光系统II水氧化酶的S4状态。并使用网站定向旋转 标记为工具，我们正在探测服务的全蛋白生化振荡器的动力学 作为大自然最简单的昼夜节律。

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

Incorporation of Ni2+, Co2+, and Selenocysteine into the Auxiliary Fe-S Cluster of the Radical SAM Enzyme HydG.

将 Ni2 、Co2 和硒代半胱氨酸掺入自由基 SAM 酶 HydG 的辅助 Fe-S 簇中。

• DOI: 10.1021/acs.inorgchem.9b01293
• 发表时间: 2019
• 期刊: Inorganic chemistry
• 影响因子: 4.6
• 作者: Rao,Guodong;Alwan,KatherineB;Blackburn,NinianJ;Britt,RDavid
• 通讯作者: Britt,RDavid

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

Bioassembly of complex iron-sulfur enzymes: hydrogenases and nitrogenases.

• DOI: 10.1016/s1874-6047(08)60224-3
• 发表时间: 2020-01-01
• 期刊: Nat. Rev. Chem.
• 作者: Britt, R. D.;Rao, G.;Tao, L.
• 通讯作者: Tao, L.