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Thioester-Assisted Sortase-A-Mediated Ligation.

Chong ZuoRuichao DingXiangwei WuYuanxia WangGuo-Chao ChuLu-Jun LiangHuasong AiZe-Bin TongJunxiong MaoQingyun ZhengTian WangZichen LiSanling LiuDemeng Sun
Published in: Angewandte Chemie (International ed. in English) (2022)
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.
Keyphrases
  • amino acid
  • protein protein
  • binding protein
  • small molecule
  • mass spectrometry
  • nitric oxide