Yang Qingyun, Zou Kun, Wu Mingxuan
Institute of Natural Sciences, Westlake Institute for Advanced Study, Hangzhou 310024, Zhejiang Province, China.
Department of Chemistry, School of Science, Westlake University, Hangzhou 310030, Zhejiang Province, China.
Org Lett. 2025 Apr 25;27(16):4158-4163. doi: 10.1021/acs.orglett.5c00568. Epub 2025 Apr 14.
Protein phosphorylation is a very important post-translational modification that regulates diverse cellular activities. In addition to classic monophosphorylation, there is also oligophosphorylation from pyrophosphorylation to polyphosphorylation. Moreover, organophosphates may modify residues such as via AMPylation and ADPylation. Although plenty of new types of protein phosphorylation have been revealed, molecular mechanisms of the biological functions are still challenging to study due to the lack of a good method to prepare proteins of interest with such site-specific PTMs. Here we report a facile method to install inorganic and organic phosphates on peptides and proteins. Vinylaryl sulfonium with an electron-withdrawing group was applied to cysteine alkylation and subsequent cyclization by γ-sulfur yielding episulfonium. This highly electrophilic intermediate was later attacked by a phosphate reagent to yield site-specifically phosphorylated cysteine peptides and proteins. As a result, this method does not require a special precursor residue on peptides/proteins or activation of phosphate reagents. In addition, this method is applicable to diverse inorganic phosphates and organophosphate. Therefore, we believe that this method will accelerate protein phosphorylation research by simple preparation of site-specific modified proteins. We also believe it offers a simple bioconjugation strategy via a phosphate linker.
蛋白质磷酸化是一种非常重要的翻译后修饰，可调节多种细胞活动。除了经典的单磷酸化外，还存在从焦磷酸化到多磷酸化的寡磷酸化。此外，有机磷酸酯可能通过氨酰化和腺苷酰化等方式修饰残基。尽管已经揭示了许多新型的蛋白质磷酸化，但由于缺乏一种制备具有此类位点特异性翻译后修饰的目标蛋白质的良好方法，其生物学功能的分子机制研究仍然具有挑战性。在此，我们报道了一种在肽和蛋白质上安装无机和有机磷酸酯的简便方法。带有吸电子基团的乙烯基芳基锍用于半胱氨酸烷基化，随后通过γ-硫进行环化生成环锍离子。这种高亲电中间体随后被磷酸试剂攻击，生成位点特异性磷酸化的半胱氨酸肽和蛋白质。因此，该方法不需要肽/蛋白质上有特殊的前体残基或磷酸试剂的活化。此外，该方法适用于多种无机磷酸酯和有机磷酸酯。因此，我们相信该方法将通过简单制备位点特异性修饰的蛋白质来加速蛋白质磷酸化研究。我们还相信它通过磷酸连接提供了一种简单的生物共轭策略。
