College of Health Science and Environmental Engineering, Shenzhen Technology University, 518118 Shenzhen, China.
College of Chemistry and Chemical Engineering, Zhongkai University of Agriculture and Engineering, 510230 Guangzhou, China.
Anal Chem. 2024 Nov 19;96(46):18377-18383. doi: 10.1021/acs.analchem.4c02215. Epub 2024 Nov 7.
Lysine residues on protein surfaces are abundant and often found in enzyme active sites, making them critical targets for studying undruggable proteins. However, the varied microenvironment surrounding lysine residues results in a wide range of p values, complicating site-specific covalent binding. In this study, we address the challenges posed by the diverse reactivity of amino side chains by modulating the amide reaction activity of heteroaromatic activated esters. By fine-tuning the type, position, and number of heteroatoms, we successfully rationalized the regulation of their amide reaction activity, leading to the design of probes for selective lysine labeling within the proteome for profiling purposes. Systematic optimization of these esters' reactivity and selectivity has yielded a series of effective probes suitable for both and cellular applications. These findings significantly enhance our understanding of protein functions and mechanisms, facilitated by the precise identification and analysis of protein labeling and profiling.
蛋白质表面的赖氨酸残基丰富，且常存在于酶的活性部位，因此成为研究不可成药蛋白的关键靶标。然而，赖氨酸残基周围变化多样的微环境导致其 p 值范围广泛，使特异性的共价结合变得复杂。在这项研究中，我们通过调节杂芳族活化酯的酰胺反应活性来解决由氨基酸侧链的多变反应性带来的挑战。通过精细调整杂原子的类型、位置和数量，我们成功地对其酰胺反应活性进行了调控，设计出用于在蛋白质组内进行选择性赖氨酸标记以进行分析的探针。对这些酯的反应性和选择性进行系统优化，得到了一系列适用于 和细胞应用的有效探针。这些发现通过对蛋白质标记和分析的精确鉴定和分析，极大地促进了我们对蛋白质功能和机制的理解。
