Functions of Carbon-Oxygen Hydrogen Bonding in Biological Methyl Transfer

生命过程化学项目通过这一奖项资助密歇根大学的Raymond Trievel教授研究甲基C-H-氧氢键在S依赖的赖氨酸甲基转移酶中起重要作用的命题。ADOMet依赖的甲基化是生物学中普遍存在的一种反应，在代谢和信号转导中起着不可或缺的作用，特别是在表观遗传基因调控中的DNA和组蛋白赖氨酸甲基化方面。据报道，非传统的C-H-O氢键可以协调赖氨酸甲基转移酶活性部位上的ADOMet甲基，但它们在甲基转移中的作用还知之甚少。以赖氨酸甲基转移酶Set7/9为模型酶，本项目将结合酶学、X射线结晶学、核磁共振波谱和非天然氨基酸突变等综合方法，研究这些氢键在ADOMet识别和催化中的功能。这些研究的结果将提供一个概念性的框架，确定碳氧氢键在赖氨酸甲基转移酶中的酶作用，以及其他类型的依赖于ADOMet的甲基转移酶也显示出甲基碳氧氢键。在更广泛的背景下，预计这项工作将提供洞察力，使人们更全面地了解C-H-O氢键在生物甲基转移中的功能重要性。在培训/教育影响方面，该项目将在化学生物学、结构生物学和生物物理学/光谱学领域吸引本科生和研究生，在化学-生物学界面提供富有启发性的多学科研究经验。国际和平协会将继续努力，将传统上代表性不足的群体的妇女和学生带入研究实验室。

With this award, the Chemistry of Life Processes Program is funding Professor Raymond Trievel at the University of Michigan to investigate the proposition that methyl C-H-oxygen hydrogen bonding plays an important role in S-adenosylmethionine (AdoMet or SAM)-dependent lysine methyltransferases. AdoMet-dependent methylation is a ubiquitous reaction in biology and has integral roles in metabolism and signal transduction, particularly with respect to DNA and histone lysine methylation in epigenetic gene regulation. Unconventional C-H---O hydrogen bonds have been reported to coordinate the AdoMet methyl group in the active sites of lysine methyltransferases, but their role in methyl transfer has remained poorly understood. Using the lysine methyltransferase SET7/9 as a model enzyme, this project will examine the functions of these hydrogen bonds in AdoMet recognition and catalysis using an integrated approach combining enzymology, X-ray crystallography, NMR spectroscopy, and unnatural amino acid mutagenesis. The results of these studies will provide a conceptual framework that defines the enzymatic roles of carbon-oxygen hydrogen bonding in lysine methyltransferases as well as other classes of AdoMet-dependent methyltransferases that also exhibit methyl carbon-oxygen hydrogen bonding. In the broader context, it is envisioned that this work will provide insights that lead to a more general understanding of the functional importance of C-H---O hydrogen bonding in biological methyl transfer. With respect to training/educational impact, this project will engage both undergraduate and graduate students in the fields of chemical biology, structural biology and biophysics/spectroscopy, providing for a stimulating, multi-disciplinary research experience at the Chemistry-Biology interface. The PI will continue to work to bring women and students from traditionally underrepresented groups into the research laboratory.