Intein-mediated protein ligation: harnessing nature's escape artists.
Inteins are naturally occurring proteins that are involved in the precise cleavage and formation of peptide bonds in a process known as protein splicing. Genetic engineering has allowed the controllable cleavage of peptide bonds at either the N- or C-terminus of the intein. Inteins displaying controllable cleavage have been used in the isolation of bacterially expressed proteins possessing either a C-terminal thioester or an N-terminal cysteine. The specific placement of these reactive groups has allowed either protein-protein or protein-peptide condensation through a native peptide bond. This review describes the methods used to specifically generate these reactive groups on bacterially expressed proteins and some applications of this technique, known as intein-mediated protein ligation. Furthermore, a versatile two intein (TWIN) system will be described which enables the circularization and polymerization of bacterially expressed proteins or peptides.
Evans
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New England Biolabs, Inc., Beverly, MA 01915-5599, USA.
Xu
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D020816:Amino Acid Motifs; D020134:Catalytic Domain; D010455:Peptides / Q000737:chemistry*; D015202:Protein Engineering / Q000379:methods; D019154:Protein Splicing
D010455:Peptides