CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
College of Chemical Engineering, Shenyang University of Chemical Technology, Shenyang 110142, China.
J Am Chem Soc. 2023 Mar 8;145(9):5252-5260. doi: 10.1021/jacs.2c12653. Epub 2023 Feb 27.
The quantitative profiling of residue reactivity in proteins promotes the discovery of covalent druggable targets for precise therapy. Histidine (His) residues, accounting for more than 20% of the active sites in enzymes, have not been systematically characterized for their reactivity, due to lack of labeling probes. Herein, we report a chemical proteomics platform for the site-specific quantitative analysis of His reactivity by combination of acrolein (ACR) labeling and reversible hydrazine chemistry enrichment. Based on this platform, in-depth characterization of His residues was conducted for the human proteome, in which the rich content of His residues (>8200) was quantified, including 317 His hyper-reactive residues. Intriguingly, it was observed that the hyper-reactive residues were less likely to be the sites for phosphorylation, and the possible mechanism of this antagonistic effect still needs to be evaluated in further research. Based on the first comprehensive map of His residue reactivity, many more residues could be adopted as the bindable sites to disrupt the activities of a diverse number of proteins; meanwhile, ACR derivatives could also be used as a novel reactive warhead in the development of covalent inhibitors.
蛋白质中残留反应性的定量分析可促进发现共价药物靶标，从而实现精准治疗。由于缺乏标记探针，占酶活性部位 20%以上的组氨酸（His）残基的反应性尚未得到系统表征。在此，我们报告了一种通过丙烯醛（ACR）标记和可逆肼化学富集相结合的化学蛋白质组学平台，用于 His 反应性的定点定量分析。基于该平台，对人蛋白质组中的 His 残基进行了深入表征，其中定量了丰富的 His 残基（>8200），包括 317 个 His 超反应性残基。有趣的是，观察到超反应性残基不太可能成为磷酸化的位点，这种拮抗作用的可能机制仍需要在进一步的研究中进行评估。基于 His 残基反应性的首个全面图谱，可以将更多的残基用作结合位点来破坏大量蛋白质的活性；同时，ACR 衍生物也可以用作开发共价抑制剂的新型反应性弹头。
