Synchronizing enzyme-mediated radical chemistry with domain motion

将酶介导的自由基化学与域运动同步

• 批准号: 386655-2010
• 负责人: Wolthers, Kirsten
• 金额: $ 2.77万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2013
• 资助国家: 加拿大
• 起止时间: 2013-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=529000
• 关键词: Synchronizing; enzyme; mediated; radical; chemistry

The research outlined in this proposal examines two relatively new and uncharted areas of enzymology: (1) radical-based reaction mechanisms and (2) the role of protein conformational motion in directing catalysis. A recent upsurge in genome sequencing and improved detection techniques has led to a rapid increase in the number of enzymes known or hypothesized to proceed by radical-based mechanisms. As many of these enzymes are key players in important biological processes, it is critical for us to understand, not only how they have evolved to control the trajectories of radicals, but also how this type of chemistry is exquisitely choreographed with varying levels and frequencies of protein motion. Using a coenzyme B12-dependent enzyme as a model system, the research program will strive to derive models for how these types of enzymes operate. The studies will compare the atomic structure of the enzyme "poised" at different states during catalysis, in order to pinpoint aspects of the protein that may play a role in radical stability and/or its dynamic properties. These structural features will be altered and the kinetic and functional behavior of the enzyme will be analyzed using spectroscopic (UV-visible and fluorescence) and fast reaction techniques. In collaboration with internationally recognized inorganic chemists (specializing in synthesis and design of radical-based catalysts) at UBC-Okanagan, novel coenzyme B12 analogs will be created that will allow us to investigate the reactive nature of the molecule. An understanding of how enzymes deploy radical-based chemistry is the first step towards engineering them to perform desired reactions that are applicable to industry and drug design, an objective that will ultimately improve the quality of life for Canadians in the future. Improved models for radical-based catalysis can also be applied to more medically relevant systems, that are often less accessible for investigation. Finally, the research will make a significant contribution to the field of enzymology, as it examines enzyme catalysis, not just in the context of "structure defines function" paradigm, but also from the emerging concept that conformational dynamics plays an integral role in its catalytic behavior.

本提案中概述的研究考察了酶学的两个相对较新和未知的领域：（1）基于自由基的反应机制和（2）蛋白质构象运动在定向催化中的作用。 最近在基因组测序和改进的检测技术的高潮，导致了已知的或假设的酶的数量迅速增加，以自由基为基础的机制进行。 由于这些酶中有许多是重要生物过程中的关键参与者，因此我们不仅要了解它们如何进化以控制自由基的轨迹，而且还要了解这种类型的化学反应是如何随着蛋白质运动的不同水平和频率而精心编排的。 使用辅酶B12依赖性酶作为模型系统，该研究计划将努力推导出这些类型的酶如何运作的模型。 这些研究将比较在催化过程中处于不同状态的酶的原子结构，以确定蛋白质可能在自由基稳定性和/或其动态特性中发挥作用的方面。这些结构特征将被改变，酶的动力学和功能行为将使用光谱（紫外可见和荧光）和快速反应技术进行分析。与UBC-Okanagan的国际公认的无机化学家（专门从事自由基催化剂的合成和设计）合作，将创建新的辅酶B12类似物，使我们能够研究分子的反应性质。了解酶如何部署基于自由基的化学是工程它们执行适用于工业和药物设计的所需反应的第一步，这一目标最终将改善加拿大人未来的生活质量。 基于自由基的催化的改进模型也可以应用于更医学相关的系统，这些系统通常不太容易进行调查。最后，这项研究将对酶学领域做出重大贡献，因为它不仅在“结构定义功能”范式的背景下研究酶催化，而且还从构象动力学在其催化行为中发挥不可或缺的作用这一新兴概念中研究酶催化。