Specific N-terminal protein labelling: use of FMDV 3C~(pro) protease and native chemical ligation

摘要

We report an effective strategy for generating N-terminal cysteinyl proteins by proteolytic cleavage using the enzyme 3C~(pro), suitable for a wide range of applications via native chemical ligation.rnNative chemical ligation (NCL) is a highly selective and facile reaction, allowing a thioester-containing moiety to be joined to an N-terminal cysteine residue via a peptide bond. This non-enzymatic, chemoselective reaction occurs at physiological pH and in aqueous solution, making it a useful tool for chemical biology; the thioester being attached can have any physicochemical property that can be tuned by synthesis. NCL is a key tool for a wide range of protein engineering applications including protein labelling and semi-synthesis of difficult to express or post-translationally modified proteins.