Thioester‐Assisted Sortase‐A‐Mediated Ligation

作者：Chong Zuo, Ruichao Ding, Xiangwei Wu, Yuanxia Wang, Guo‐Chao Chu, Lujun Liang, Huasong Ai, Zebin Tong, Junxiong Mao, Qingyun Zheng, Tian Wang, Zichen Li, Lei Liu, Demeng Sun · 发表于：Angewandte Chemie International Edition · 年份：2022 · DOI：10.1002/anie.202201887 · 被引用次数：51 · 研究领域：Biochemical and Structural Characterization、Glycosylation and Glycoproteins Research、Chemical Synthesis and Analysis

Sortase A (SrtA)-mediated ligation, a popular method for protein labeling and semi-synthesis, is limited by its reversibility and dependence on the LPxTG motif, where "x" is any amino acid. Here, we report that SrtA can mediate the efficient and irreversible ligation of a protein/peptide containing a C-terminal thioester with another protein/peptide bearing an N-terminal Gly, with broad tolerance for a wide variety of LPxT-derived sequences. This strategy, the thioester-assisted SrtA-mediated ligation, enabled the expedient preparation of proteins bearing various N- or C-terminal labels, including post-translationally modified proteins such as the Ser139-phosphorylated histone H2AX and Lys9-methylated histone H3, with less dependence on the LPxTG motif. Our study validates the chemical modification of substrates as an effective means of augmenting the synthetic capability of existing enzymatic methods.