NMR Crystallography: Linking Chemical Structure and Mechanism in Tryptophan Synthase

NMR 晶体学：连接色氨酸合酶的化学结构和机制

• 批准号: 1710671
• 负责人: Leonard Mueller
• 金额: $ 48万
• 依托单位: University of California-Riverside
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1710671&HistoricalAwards=false
• 关键词: NMR; Crystallography; Linking; Chemical; Structure

This award is funded by the Chemistry Division's Chemistry of Life Processes Program. Professors Leonard Mueller and Li Fan from the University of California at Riverside bring together two of the tools of molecular imaging, nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography, to describe an enzymatic reaction with atomic resolution (the ability to distinguish between atoms of a molecule and between molecules). Enzymes catalyze specific chemical reactions that impact nearly all life processes. Yet the details of how they do this remain elusive. In this study, the location of all atoms including hydrogens, the smallest of all atoms, are described as the reaction takes place. The result is a unique and detailed view into the reaction that shows how atoms move during the process. This new method is applied to one enzyme in particular, but the basic approach should be widely applicable to many enzymes. This research is carried out by graduate and undergraduate students at UC Riverside and by first-generation undergraduate students from neighboring Claremont colleges. The researchers share their results and experience at scientific conferences and in the local elementary and high schools.This project develops of NMR-assisted crystallography, the synergistic combination of solid-state NMR spectroscopy, X-ray crystallography, and computational chemistry as an atomic-resolution probe of structure and dynamics in enzyme active sites. The goal is to understand the molecular basis for reaction specificity in the pyridoxal-5'-phosphate (PLP) dependent enzyme tryptophan synthase (TS). To accomplish this goal, Professors Mueller and Fan describe with atomic resolution, the transformation of substrates into product in the beta-subunit active site of TS. The location of all atoms is specified, including hydrogens, for multiple intermediates along the beta-reaction pathway. The combination of X-ray crystallography, solid-state NMR, and computational methodologies give chemically-rich detail that is not available when these methodologies are applied in isolation. Taken together, these techniques can provide consistent and testable models for structure and function in an enzyme active site: X-ray crystallography provides a coarse framework upon which models of the active site can be developed using computational chemistry. These models can be tested by comparisons of their first-principles predicted chemical shifts and the results of solid-state NMR spectroscopy experiments.

该奖项由化学部的生命过程化学计划资助。来自滨江的加州大学的伦纳德·穆勒教授和李凡教授将两种分子成像工具--核磁共振（NMR）光谱学和X射线晶体学--结合在一起，以原子分辨率（区分分子原子和分子之间的能力）描述酶促反应。酶催化影响几乎所有生命过程的特定化学反应。然而，他们如何做到这一点的细节仍然难以捉摸。在这项研究中，所有原子（包括所有原子中最小的氢原子）的位置都被描述为反应发生的位置。其结果是对反应的独特而详细的看法，显示了原子在过程中如何移动。这种新方法特别适用于一种酶，但基本方法应该广泛适用于许多酶。这项研究是由研究生和本科生在加州大学滨江和第一代本科生从邻近的克莱蒙学院。研究人员在科学会议和当地中小学分享了他们的成果和经验。该项目开发了核磁共振辅助晶体学，即固态核磁共振光谱学、X射线晶体学和计算化学的协同组合，作为结构的原子分辨率探针和酶活性位点的动力学。目的是了解吡哆醛-5 '-磷酸（PLP）依赖性酶色氨酸合酶（TS）反应特异性的分子基础。为了实现这一目标，Mueller和Fan教授以原子分辨率描述了TS的β亚基活性位点中底物转化为产物的过程。所有原子的位置都被指定，包括氢，用于沿着β-反应途径的多个中间体沿着。X射线晶体学、固态NMR和计算方法的结合提供了化学上丰富的细节，这些细节在孤立地应用这些方法时是不可用的。总之，这些技术可以提供一致的和可测试的模型的结构和功能的酶的活性位点：X射线晶体学提供了一个粗略的框架上，模型的活性位点可以使用计算化学开发。 这些模型可以通过比较它们的第一性原理预测的化学位移和固态NMR光谱实验的结果进行测试。

项目成果

Backbone assignments and conformational dynamics in the S. typhimurium tryptophan synthase α-subunit from solution-state NMR

溶液态 NMR 中鼠伤寒沙门氏菌色氨酸合酶 α 亚基的主链分配和构象动力学

• DOI: 10.1007/s10858-020-00320-2
• 发表时间: 2020
• 期刊: Journal of Biomolecular NMR
• 影响因子: 2.7
• 作者: Sakhrani, Varun V.;Hilario, Eduardo;Caulkins, Bethany G.;Hatcher-Skeers, Mary E.;Fan, Li;Dunn, Michael F.;Mueller, Leonard J.
• 通讯作者: Mueller, Leonard J.

Cofactor-Mediated Nucleophilic Substitution Catalyzed by a Self-Assembled Holoenzyme Mimic

• DOI: 10.1021/acs.joc.9b01880
• 发表时间: 2019-09-20
• 期刊: JOURNAL OF ORGANIC CHEMISTRY
• 影响因子: 3.6
• 作者: Ngai, Courtney;Bogie, Paul M.;Hooley, Richard J.
• 通讯作者: Hooley, Richard J.

Non-Uniform Sampling in NMR Spectroscopy and the Preservation of Spectral Knowledge in the Time and Frequency Domains

• DOI: 10.1021/acs.jpca.0c02930
• 发表时间: 2020-07-02
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY A
• 影响因子: 2.9
• 作者: Kaur，Manpreet;Lewis，Callie M.;Mueller，Leonard J.
• 通讯作者: Mueller，Leonard J.