Chemical Probes to Study Methionine Redox Biology

项目摘要/摘要 蛋白质蛋氨酸残基正在成为跨越信号的生理过程的重要氧化还原位点 转导、抗氧化防御和蛋白质功能的调节。与此同时，这里异常的海拔高度 氧化应激和/或蛋氨酸亚砜还原酶失活引起的蛋氨酸氧化 酶可导致神经退行性疾病和血管疾病、寿命和癌症进展。至 帮助破译蛋氨酸氧化还原生理学和病理学的贡献，我们正在开发新的化学物质 选择性标记蛋白质中反应性蛋氨酸位点的试剂。科学的前提是提供 获得蛋氨酸生物偶联的模块化化学工具箱将有助于揭开新功能蛋氨酸的面纱 蛋白质中的位点进一步了解蛋氨酸氧化的生理和病理作用 和减少。利用模拟天然氧化的氧化还原探针选择性地进行蛋氨酸生物偶联 这些残基的化学提供了化学创新，同时通过 组合使用不同的和互补的合成、建模、生化、蛋白质组和成像 为探索反应性蛋氨酸景观提供通用工具的方法。具体地说，我们寻求(1) 通过建立一种预测模型来设计和合成强健且高度特异的恶氮杂环丙啶探针 调整蛋氨酸加合物的选择性和稳定性，(2)确定氧化敏感的蛋氨酸位点和 化学蛋白质组对参与可逆氧化还原调控的特定蛋白质靶点的生化表征 在蛋白质组上比较有或没有蛋氨酸亚砜还原酶擦除蛋白的细胞模型。 以及(3)开发基于邻近连接分析的新的成像探针来监测蛋氨酸- 细胞中选定蛋白质靶标的依赖氧化还原状态。

Project Summary/Abstract Protein methionine residues are emerging as essential redox sites for physiological processes spanning signal transduction, antioxidant defense, and regulation of protein function. At the same time, aberrant elevations in methionine oxidation induced by oxidative stress and/or inactivation of methionine sulfoxide reductase enzymes can contribute to neurodegenerative and vascular diseases, life span, and cancer progression. To help decipher contributions of methionine redox physiology and pathology, we are developing new chemical reagents to selectively label reactive methionine sites in proteins. The scientific premise is that providing access to a modular chemical toolbox for methionine bioconjugation will help unveil new functional methionine sites in proteins to further understanding of physiological and pathological contributions of methionine oxidation and reduction. Selective methionine bioconjugation using redox-based probes that emulate the innate oxidative chemistry of these residues offers chemical innovation, while demonstrating scientific rigor through the combined use of diverse and complementary synthetic, modeling, biochemical, proteomic, and imaging approaches to provide general tools for probing the reactive methionine landscape. Specifically, we seek to (1) design and synthesize robust and highly specific oxaziridine probes by establishing a predictive model for tuning selectivity and stability of methionine adducts, (2) identify oxidatively-sensitive methionine sites and biochemically characterize specific protein targets involved in reversible redox regulation by chemoproteomic comparison of cell models with or without the methionine sulfoxide reductase eraser protein on a proteome- wide scale, and (3) develop new imaging probes based on proximity ligation assays to monitor methionine- dependent redox status of selected protein targets in cells.