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.

金属是所有蛋白质的大约50％的重要组成部分，其功能从储能和代谢到分子转运和细胞信号传导不等。 但是，由于这些金属蛋白系统的复杂性（其中许多都包含多个不同的金属中心）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，以更好地了解这些金属蛋白如何发挥作用，从而开发一种研究这些反应的新方法。 该方法将应用于自然和工程酶中氢的生产和氧化的研究，并在替代能源，可持续燃料，二氧化碳固定和氮气循环等人等中使用未来的潜在应用。 此外，该提案的研究目标与教育和外展活动融合在一起，对科学和社会都有深远的影响。 涉及生物学，化学和物理学方面的工作的跨学科性质，使受众群体可以接受。 该教育计划将以沉浸式的方式揭示从代表性不足的种族/族裔群体，残疾人到现代化学研究的广泛学生，目的是建立一种以职业为导向的STEM人才管道。 将向成人受众的一般利益研讨会提出，以提高科学素养，提高意识，更好地吸引社区参与OSU和世界各地的当代研究。与俄亥俄州大学的化学过程中的化学过程中，这一奖项是从俄亥俄州立大学中的化学过程中的，从而在俄亥俄州大学进行了启用，从而在俄亥俄州大学进行了研究。光谱法（CGRR）。 通过利用在高度可调的，最先进的仪器上进行的共振拉曼光谱的选择性和敏感性，将获得有关催化中间体结构的重要信息，包括质子化状态，键盘顺序和协调模式。 与光谱特性的理论预测结合的位点选择性和特定于状态的激发将用于表征传统方法无法访问的中间体和瞬时物种。 将研究金属酶活性位点的小分子活化反应，包括质子还原，氢氧化和氢化酶和模型的氧还原，以证明这种实验计算方法的实用性并鉴定催化过程中的关键结构基序。这种协同方法将广泛地适用于许多类别的酶，尤其是含有多种金属生物活性剂的酶，例如氮气和一氧化碳脱氢酶。 使用CGRR，将有可能获得分子级别的洞察力，并开始了解金属蛋白如何作为已知的一些最有效的催化剂作用。

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.