Department of Chemistry, University of Virginia, Charlottesville, VA, USA.
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA, USA.
Nat Chem Biol. 2020 Feb;16(2):150-159. doi: 10.1038/s41589-019-0404-5. Epub 2019 Nov 25.
Covalent probes serve as valuable tools for global investigation of protein function and ligand binding capacity. Despite efforts to expand coverage of residues available for chemical proteomics (e.g., cysteine and lysine), a large fraction of the proteome remains inaccessible with current activity-based probes. Here, we introduce sulfur-triazole exchange (SuTEx) chemistry as a tunable platform for developing covalent probes with broad applications for chemical proteomics. We show modifications to the triazole leaving group can furnish sulfonyl probes with ~5-fold enhanced chemoselectivity for tyrosines over other nucleophilic amino acids to investigate more than 10,000 tyrosine sites in lysates and live cells. We discover that tyrosines with enhanced nucleophilicity are enriched in enzymatic, protein-protein interaction and nucleotide recognition domains. We apply SuTEx as a chemical phosphoproteomics strategy to monitor activation of phosphotyrosine sites. Collectively, we describe SuTEx as a biocompatible chemistry for chemical biology investigations of the human proteome.
共价探针可用于全面研究蛋白质功能和配体结合能力，是一种非常有价值的工具。尽管人们一直在努力扩大可用于化学蛋白质组学的残基覆盖范围（例如半胱氨酸和赖氨酸），但目前基于活性的探针仍然无法检测到很大一部分蛋白质组。在这里，我们引入了硫三唑交换（SuTEx）化学作为一种可调谐平台，用于开发具有广泛化学蛋白质组学应用的共价探针。我们发现，通过改变三唑离去基团，可以为磺酰探针提供约 5 倍的酪氨酸化学选择性，从而在裂解物和活细胞中研究超过 10000 个酪氨酸位点。我们发现，具有增强亲核性的酪氨酸富集在酶、蛋白质-蛋白质相互作用和核苷酸识别结构域中。我们将 SuTEx 应用于化学磷酸蛋白质组学策略来监测磷酸酪氨酸位点的激活。总的来说，我们将 SuTEx 描述为一种用于人类蛋白质组学的生物相容性化学生物学研究的化学方法。
