Investigation of Two New Protein Post-Translational Modifications Derived from a Key Metabolite in Valine Metabolism

缬氨酸代谢中关键代谢物衍生的两种新蛋白质翻译后修饰的研究

• 批准号: 2203942
• 负责人: Y George Zheng
• 金额: $ 49.46万
• 依托单位: University of Georgia Research Foundation Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-15 至 2026-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2203942&HistoricalAwards=false
• 关键词: Investigation; Two; New; Protein; Post; Investigation; Two; New; Protein; Post

With the support of the Chemistry of Life Processes (CLP) Program in the Division of Chemistry, Prof Y. George Zheng of the University of Georgia is investigating new molecular mechanisms of life process regulation by novel post-translational modification of proteins. Specific chemical modifications in proteins can impact key attributes and functions of affected proteins, such as structural conformation, enzymatic activity, intracellular localization, and interaction with other biomolecules. Through fine-tuned post-translational modifications, cells and organisms adjust and regulate their physiological states in response to various internal and external stimuli. Currently, the molecular mechanisms of many post-translational modifications and their roles in regulating biological processes are incompletely understood. This also highlights another key point; namely that there are likely a good number of post-translational modifications (PTMs) yet to be characterized. This project endeavors to advance biological research field by identifying as yet unrecognized post-translational modifications and elucidating the mechanisms by which several of these PTMs regulate key life processes such as DNA transcription, cell cycle control, and signal transduction. Multidisciplinary methods and tools in the chemical and biological sciences will be explored and applied to identify, validate, and functionally annotate post-translational modifications in the human proteome. The proposed work is expected to support education and diversity, consistent with the the principal investigator’s established record in the scientific training of graduate, undergraduate and high school students from diverse backgrounds. Early career trainees will learn and apply state-of-the-art methods in chemical biology to address relevant questions in molecular signaling, thereby benefiting the chemical biology science community broadly.In this project, a particular focus will be placed on identifying and validating two new post-translational modifications in human proteins that are derived from the key metabolites in the valine metabolic pathway. The researchers hypothesize that the reactive metabolite, methacrylyl-CoA, and/or its hydrolyzed product methacrylate, produced in valine metabolism are capable of modifying cellular proteins through two different biochemical mechanisms: acylation of specific lysine residues (lysine methacrylylation) and alkylation of specific cysteine residues (S-2-carboxypropylation). Tailored chemical probes will be developed to specifically recognize and label lysine methacrylylation and cysteine S-2-carboxypropylation molecular modifications in proteins and peptides, and then be applied to set up chemoproteomic platforms to map out methacrylylated and S-2-carboxypropylated protein substrates at the proteomic level. Bioinformatics will be used to delineate the distribution and scope of lysine methacrylylation and cysteine S-2-carboxypropylation in the human proteome. Furthermore, cellular and biochemical experiments will be conducted to investigate dynamic controls and downstream effects of these protein modifications. This work is projected to expand understanding of valine metabolites and their contribution to protein posttranslational modification, and to a broader level, advance the tool kit for molecular-level understanding of the expressed human proteome and the role of PTMs in regulation and signaling.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学过程中的化学过程（CLP）方案的支持下，佐治亚大学的乔治·郑教授正在研究通过蛋白质的新型翻译后修饰来调节生命过程的新分子机制。蛋白质中的特定化学修饰会影响受影响蛋白质的关键属性和功能，例如结构构象，酶活性，细胞内定位以及与其他生物分子的相互作用。通过微调的翻译后修饰，细胞和生物会根据各种内部和外部刺激来调整和调节其身体状态。目前，尚不完全了解许多翻译后修饰的分子机制及其在受调节的生物过程中的作用。这也突出了另一个关键点。也就是说，通过鉴定尚未识别的翻译后修饰，并阐明这些PTM的几个调节诸如DNA转录，细胞周期控制和信号转导的机制，可能会有大量的翻译后修饰（PTMS）这项项目努力来推进生物学研究领域。将探索并应用化学​​和生物科学中的多学科方法和工具，以识别，验证和功能上注释人类蛋白质组的翻译后修饰。预计拟议的工作将支持教育和多样性，这与主要研究人员在科学培训中的既定记录都在研究生，本科和高中生的科学培训方面保持一致。早期的职业学员将学习和应用化学生物学的最先进方法来解决分子信号中的相关问题，从而使化学生物学科学界受益匪浅。在该项目中，将特别着重于识别和验证两种新的在人类蛋白质中的新的翻译后修饰，这些蛋白质源自Valine Nerabolic Path中的关键化合物。研究人员假设，在瓣膜代谢中产生的反应性代谢物，甲基丙烯酸-COA和/或其水解产物甲基丙烯酸酯能够通过两种不同的生化机制来修饰细胞蛋白：特定赖氨酸残基的酰基化（裂解甲基甲基丙烯酸酯）的酰基化（裂解的甲基苯胺）的酶促化。 （S-2羧基化）。量身定制的化学问题将被开发出来，以专门识别和标记赖氨酸甲基酰基化和半胱氨酸S-2-羧基化分子在蛋白质和辣椒中的修饰，然后应用于在该蛋白质蛋白质级别的蛋白质蛋白质级别绘制化学蛋白质疗法平台，以绘制甲基甲基甲基化和S-2-羧基蛋白质的蛋白质水平。生物信息学将用于描述人蛋白中赖氨酸甲基酰基化和半胱氨酸S-2-羧基化的分布和范围。此外，将进行细胞和生化实验，以研究这些蛋白质修饰的动态控制和下游效应。预计这项工作旨在扩大对门代谢产物的理解及其对蛋白质后翻译后修改的贡献，并在更广泛的水平上提高了工具包，以了解分子级别对表达的人蛋白质组的理解，以及PTM在调节和信号中的作用。这种奖励通过评估官员的范围来表现出良好的依据。