School of Chemistry and Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Sydney, Sydney, NSW 2006, Australia.
Faculty of Chemistry, Institute of Biological Chemistry, University of Vienna, Vienna, Austria.
Angew Chem Int Ed Engl. 2022 May 9;61(20):e202200163. doi: 10.1002/anie.202200163. Epub 2022 Mar 9.
Herein, we describe the development and application of a novel expressed protein selenoester ligation (EPSL) methodology for the one-pot semi-synthesis of modified proteins. EPSL harnesses the rapid kinetics of ligation reactions between modified synthetic selenopeptides and protein aryl selenoesters (generated from expressed intein fusion precursors) followed by in situ chemoselective deselenization to afford target proteins at concentrations that preclude the use of traditional ligation methods. The utility of the EPSL technology is showcased through the efficient semi-synthesis of ubiquitinated polypeptides, lipidated analogues of the membrane-associated GTPase YPT6, and site-specifically phosphorylated variants of the oligomeric chaperone protein Hsp27 at high dilution.
在此，我们描述了一种新颖的表达蛋白硒酯连接（EPSL）方法的开发和应用，用于修饰蛋白的一锅半合成。EPSL 利用修饰后的合成硒肽与蛋白芳基硒酯（由表达的内含肽融合前体生成）之间的快速连接反应动力学，然后进行原位化学选择性脱硫，以在排除传统连接方法使用的浓度下获得目标蛋白。通过高效半合成泛素化多肽、膜相关 GTP 酶 YPT6 的脂化类似物以及寡聚伴侣蛋白 Hsp27 的定点磷酸化变体，展示了 EPSL 技术的实用性。
