中国科学技术大学Sun Demeng团队报道了硫酯辅助的Sortase-A介导的连接。相关研究成果发表在2022年5月6日出版的《德国应用化学》。

Sortase A（SrtA）介导的连接是一种常用的蛋白质标记和半合成方法，但由于其可逆性和对LPxTG基序的依赖性而受到限制，其中“x”是任何氨基酸。

该文中，研究人员报告SrtA可以介导一种含有C端硫酯的蛋白质/肽与另一种含有N端甘氨酸的蛋白质/肽的高效不可逆连接，对多种LPxT衍生序列具有广泛的耐受性。该策略，即硫酯辅助SrtA介导的连接，能够方便地制备带有各种N-或C-末端标签的蛋白质，包括翻译后修饰的蛋白质，如Ser139磷酸化组蛋白H2AX和Lys9甲基化组蛋白H3，对LPxTG基序的依赖性较小。该研究证实了底物的化学修饰是增强现有酶法合成能力的有效手段。

附：英文原文

Title: Thioester-Assisted Sortase-A - Mediated Ligation

Author: Chong Zuo, Ruichao Ding, Xiangwei Wu, Yuanxia Wang, Guo-Chao Chu, Lu-Jun Liang, Huasong Ai, Ze-Bin Tong, Junxiong Mao, Qingyun Zheng, Tian Wang, Zichen Li, Lei Liu, Demeng Sun

Issue&Volume: 2022-05-06

Abstract: 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.

DOI: 10.1002/anie.202201887

Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202201887