Advanced Spectroscopic and Computational Analysis of Metal Sites in Enzymes, Biomimetic Models, and Catalytic Intermediates.

酶中金属位置的高级光谱分析和计算分析、仿生模型和 催化中间体。 摘要/摘要 包括人类在内的所有生物体的生命都依赖于 金属蛋白提供选择性和快速的化学转化。建议的目标是 研究是通过监测和分析原子来阐明特定金属酶是如何发挥作用的 在反应过程中金属活性中心发生的能级变化。这些研究还补充了 仿生复合体的基准研究。从这项工作中收集到的理解将提供一个 寻找疾病病因和治疗方法的分子基础。研究工作的重点将放在 金属蛋白中活性中心与蛋白质基质的相互作用如何影响酶 功能。原子对金属的初级配位至关重要，但在许多情况下，配位作用较弱。 次级球体相互作用，通常是来自附近氨基酸残基的氢键 也产生了重大影响。我们实验室的研究使用了先进的光谱和计算技术 方法，提供可比较的蛋白质中金属活性部位的详细特征 我们和其他研究人员从合成材料中收集了大量基准数据库 多年来的模型综合体。为了了解酶的工作原理，我们研究了它们的关键步骤 捕捉、表征活性中间体及其元素示踪的化学机理 毫秒时间尺度上的动力学。这项工作是高度协作的，因为我们依赖于 许多合成和生化研究小组，以确保获得仿生和蛋白质复合体 通过共同努力，可以用定义明确的协议干净地准备好这一点。

Advanced Spectroscopic and Computational Analysis of Metal Sites in Enzymes, Biomimetic Models, and Catalytic Intermediates. Summary/Abstract Life of all organisms, including humans, depends on the activation of small stable molecules by metalloproteins to provide selective and rapid chemical transformations. The goal of the proposed research is to elucidate how specific metalloenzymes function by monitoring and analyzing the atomic level changes that occur at the metal active site during the reaction. These studies are augmented with benchmark studies of biomimetic complexes. The understanding gleaned from this work will provide a molecular basis for finding causes and remedies of diseases. The focus of the research efforts will be on how the interfacing of the active sites in metalloproteins with the protein matrix affects enzymatic function. The primary atomic coordination to the metal is of critical importance, but in many cases, weaker secondary sphere interactions, usually hydrogen bonds from nearby amino acid residues can have significant influence as well. The investigations in our lab use advanced spectroscopic and computational methods, providing detailed characterizations of the metal active sites in proteins that can be compared with an extensive database of benchmarks that we and other researchers have gathered from synthetic model complexes over the years. To understand how an enzyme works, we study key steps in their chemical mechanism by trapping and characterizing reactive intermediates and tracking their elemental kinetics on a millisecond time scale. The work is highly collaborative as we depend on the expertise of many synthetic and biochemical research groups to ensure access to biomimetic and protein complexes that can be, through joint effort, prepared cleanly with well-defined protocols.