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，作用机制往往仍然难以捉摸。 这项研究将通过将实验室实验与计算化学相结合来研究这些反应，以更好地了解这些金属蛋白的功能。 这种方法将被应用于天然和工程酶的氢生产和氧化的研究，未来可能应用于替代能源，可持续燃料，二氧化碳固定和氮循环等。 此外，该建议的研究目标与教育和外联活动相结合，对科学和社会都有深远的影响。 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，使该主题能够为各种各样的观众所接受。 该教育计划将使来自代表性不足的种族/民族群体和残疾学生以身临其境的方式接触现代化学研究，其目标是建立一个多元化，以职业为导向的STEM人才管道。 将为成人观众举办公益研讨会，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。凭借生命过程化学计划的这一奖项，在化学系，来自俄亥俄州州立大学的Hannah Shafaat博士将使用计算引导共振拉曼光谱（CGRRS）研究金属蛋白的酶机制。. 通过利用共振拉曼光谱的选择性和灵敏度，在一个高度可调的，国家的最先进的仪器上进行，催化中间体的结构，包括质子化状态，键级，和配位模式的重要信息，将获得。 结合光谱特性的理论预测，站点选择性和特定状态的激发将用于表征传统方法无法访问的中间体和瞬态物种。 在金属酶活性位点的小分子活化反应，包括质子还原，氢氧化，和氢酶和模型中的氧还原，将被研究，以证明这种实验-计算方法的实用性，并确定在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，特别是那些含有多种金属辅因子，如固氮酶和一氧化碳脱氢酶的研究。 使用CGRRS，将有可能获得对复杂反应机制的分子水平的洞察，并开始了解金属蛋白质如何能够作为已知的一些最有效的催化剂发挥作用。

项目成果

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
• 期刊: ACS Catalysis
• 影响因子: 12.9
• 作者: Slater, Jeffrey W.;Marguet, Sean C.;Gray, Michelle E.;Monaco, Haleigh A.;Sotomayor, Marcos;Shafaat, Hannah S.
• 通讯作者: Shafaat, Hannah S.

Heterobimetallic Models of the [NiFe] Hydrogenases: A Structural and Spectroscopic Comparison

[NiFe] 氢化酶的异双金属模型：结构和光谱比较

• DOI: 10.1080/02603594.2015.1108914
• 发表时间: 2015
• 期刊: Comments on Inorganic Chemistry
• 影响因子: 5.4
• 作者: Behnke, Shelby L.;Shafaat, Hannah S.
• 通讯作者: Shafaat, Hannah S.

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
• 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.

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
• 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. Alex;Shafaat, Hannah S.
• 通讯作者: Shafaat, Hannah S.