CAREER: Metalloenzyme mechanisms probed by resonance Raman spectroscopy

职业：通过共振拉曼光谱探测金属酶机制

• 批准号: 1454289
• 负责人: Hannah Shafaat
• 金额: $ 65万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2021-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1454289&HistoricalAwards=false
• 关键词: CAREER; Metalloenzyme; mechanisms; probed; resonance

Metals are an essential component of approximately 50% of all proteins, with functions ranging from energy storage and metabolism to molecular transport and cellular signaling. However, due to the complexity of these metalloprotein systems, many of which contain multiple, distinct metal centers, the mechanisms of action often remain elusive. This research will develop a novel approach towards studying these reactions by combining laboratory experiments with computational chemistry to better understand how these metalloproteins function. This methodology will be applied to the study of hydrogen production and oxidation in natural and engineered enzymes, with future potential applications for alternative energy, sustainable fuels, carbon dioxide fixation, and nitrogen cycling, among others. Furthermore, the research objectives of this proposal are integrated with education and outreach activities, with far-reaching impact for both science and society. The interdisciplinary nature of the work, involving aspects of biology, chemistry, and physics, renders the subject accessible to a wide variety of audiences. The educational plan will expose a broad range of students from underrepresented racial/ethnic groups and with disabilities to modern chemistry research in an immersive fashion, with the objective of building a diverse, career-oriented pipeline of STEM talent. General-interest seminars for adult audiences will be presented with aims of improving scientific literacy, increasing awareness, and better engaging the community in contemporary research being carried out at OSU and across the world.With this award from the Chemistry of Life Processes Program in the Division of Chemistry, Dr. Hannah Shafaat from The Ohio State University will study enzymatic mechanisms in metalloproteins using computationally-guided resonance Raman spectroscopy (CGRRS). By harnessing the selectivity and sensitivity of resonance Raman spectroscopy performed on a highly tunable, state-of-the-art instrument, important information on the structures of catalytic intermediates, including protonation state, bond order, and coordination mode, will be obtained. Site-selective and state-specific excitation in conjunction with theoretical predictions of spectroscopic properties will be used to characterize intermediates and transient species that are inaccessible by conventional approaches. Small molecule activation reactions at metalloenzyme active sites, including proton reduction, hydrogen oxidation, and oxygen reduction in hydrogenases and models, will be investigated to demonstrate the utility of this experimental-computational approach and identify key structural motifs during catalysis. This synergistic methodology will be broadly applicable to the study of many classes of enzymes, particularly those containing multiple metallocofactors, such as nitrogenase and carbon monoxide dehydrogenase. Using CGRRS, it will be possible to gain molecular-level insight into complex reaction mechanisms and begin to understand how metalloproteins are able to function as some of the most efficient catalysts known.

金属是大约 50% 的蛋白质的重要​​组成部分，其功能范围从能量储存和代谢到分子运输和细胞信号传导。 然而，由于这些金属蛋白系统的复杂性，其中许多包含多个不同的金属中心，其作用机制通常仍然难以捉摸。 这项研究将通过将实验室实验与计算化学相结合来开发一种研究这些反应的新方法，以更好地了解这些金属蛋白的功能。 该方法将应用于天然酶和工程酶中氢的产生和氧化的研究，以及未来在替代能源、可持续燃料、二氧化碳固定和氮循环等方面的潜在应用。 此外，该提案的研究目标与教育和推广活动相结合，对科学和社会产生了深远的影响。 这项工作的跨学科性质，涉及生物学、化学和物理学的各个方面，使该主题能够为广泛的受众所接受。 该教育计划将以身临其境的方式让来自代表性不足的种族/族裔群体和残疾的广大学生接触现代化学研究，目标是建立多元化、以职业为导向的 STEM 人才管道。 将为成人观众举办普遍感兴趣的研讨会，旨在提高科学素养、提高认识，并更好地让社区参与俄勒冈州立大学和世界各地正在进行的当代研究。俄亥俄州立大学化学系的 Hannah Shafaat 博士将利用计算引导的共振拉曼光谱 (CGRRS) 研究金属蛋白的酶促机制。 通过利用在高度可调、最先进的仪器上进行的共振拉曼光谱的选择性和灵敏度，将获得有关催化中间体结构的重要信息，包括质子化状态、键序和配位模式。 位点选择性和特定状态激发与光谱特性的理论预测相结合，将用于表征传统方法无法获得的中间体和瞬态物质。 将研究金属酶活性位点的小分子活化反应，包括氢化酶和模型中的质子还原、氢氧化和氧还原，以证明这种实验计算方法的实用性并确定催化过程中的关键结构基序。这种协同方法将广泛适用于许多类别的酶的研究，特别是那些含有多种金属辅因子的酶，例如固氮酶和一氧化碳脱氢酶。 使用 CGRRS，将有可能在分子水平上深入了解复杂的反应机制，并开始了解金属蛋白如何能够充当已知的最有效的催化剂。

项目成果

Robust Carbon-Based Electrodes for Hydrogen Evolution through Site-Selective Covalent Attachment of an Artificial Metalloenzyme

通过人工金属酶的位点选择性共价连接析氢的鲁棒碳基电极

• DOI: 10.1021/acsaem.0c02069
• 发表时间: 2020-11
• 期刊: ACS Applied Energy Materials
• 影响因子: 6.4
• 作者: Treviño, Regina E.;Slater, Jeffrey W.;Shafaat, Hannah S.
• 通讯作者: Shafaat, Hannah S.

Power of the Secondary Sphere: Modulating Hydrogenase Activity in Nickel-Substituted Rubredoxin

第二球体的力量：调节镍取代红红还蛋白中的氢化酶活性

• DOI: 10.1021/acscatal.9b01720
• 发表时间: 2019-08-02
• 期刊: ACS Catalysis
• 影响因子: 12.9
• 作者: J. W. Slater;S. Marguet;M. Gray;H. Monaco;M. Sotomayor;H. Shafaat
• 通讯作者: H. Shafaat

Protein-based models offer mechanistic insight into complex nickel metalloenzymes

基于蛋白质的模型提供了对复杂镍金属酶的机制洞察

• DOI: 10.1016/j.cbpa.2021.102110
• 发表时间: 2022-04
• 期刊: Current Opinion in Chemical Biology
• 影响因子: 7.8
• 作者: Treviño, Regina E.;Shafaat, Hannah S.
• 通讯作者: Shafaat, Hannah S.

In Vivo Assembly of a Genetically Encoded Artificial Metalloenzyme for Hydrogen Production

用于制氢的基因编码人工金属酶的体内组装

• DOI: 10.1021/acssynbio.1c00177
• 发表时间: 2021-08
• 期刊: ACS Synthetic Biology
• 影响因子: 4.7
• 作者: Naughton, Kassandra J.;Treviño, Regina E.;Moore, Peter J.;Wertz, Ashlee E.;Dickson, J. Ale;Shafaat, Hannah S.
• 通讯作者: Shafaat, Hannah S.

Uncovering Redox Non-innocent Hydrogen-Bonding in Cu(I)-Diazene Complexes

揭示 Cu(I)-二氮烯配合物中的氧化还原非无害氢键

• DOI: 10.1021/jacs.1c04108
• 发表时间: 2021-10
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Gardner, Evan J.;Marguet, Sean C.;Cobb, Caitlyn R.;Pham, Dominic M.;Beringer, Josalyne A.;Bertke, Jeffery A.;Shafaat, Hannah S.;Warren, Timothy H.
• 通讯作者: Warren, Timothy H.