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Published on: August 1, 2018
Improvement of Transpeptidative Protein Ligation via Selective Nucleophile Quenching using
Junyou Chen1, Junjiang Li1, Tingting Cui1
1School of Food and Biological Engineering, Engineering Research Center of Bio-process, Ministry of Education, Key Laboratory of Animal Source of Anhui Province, Hefei University of Technology, Hefei 230009, China.
Abstract:
Here, we report a new method that employs a simple and readily available small molecule, 2-cyano-6-hydroxybenzothiazole (CBT), to selectively quench the nucleophilicity of an N-terminal Cys-containing leaving group, thereby shifting the equilibrium of peptidyl asparaginyl ligase (PAL)-mediated ligation toward the product side. This method can be applied not only to the N- and C-terminal modifications of proteins but also to efficiently mediate protein-protein ligation.

