title: Ligation of synthetic peptides to proteins using semisynthetic protein trans-splicing.
abstract: Protein trans-splicing using split inteins is a powerful and convenient reaction to chemically modify recombinantly expressed proteins under mild conditions. In particular, semisynthetic protein trans-splicing with one intein fragment short enough to be accessible by solid-phase peptide synthesis can be used to transfer a short peptide segment with the desired synthetic moiety to the protein of interest. In this chapter, we provide detailed protocols for two such split intein systems. The M86 mutant of the Ssp DnaB intein and the MX1 mutant of the AceL-TerL intein are two highly engineered split inteins with very short N-terminal intein fragments of only 11 and 25 amino acids, respectively, and allow the efficient N-terminal labeling of proteins.
authors.affiliation: Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.
authors.forename: Julian C J
authors.initials: JC
authors.lastname: Matern
authors.forename: Anne-Lena
authors.initials: AL
authors.lastname: Bachmann
authors.forename: Ilka V
authors.initials: IV
authors.lastname: Thiel
authors.forename: Gerrit
authors.initials: G
authors.lastname: Volkmann
authors.forename: Alexandra
authors.initials: A
authors.lastname: Wasmuth
authors.forename: Jens
authors.initials: J
authors.lastname: Binschik
authors.forename: Henning D
authors.initials: HD
authors.lastname: Mootz
mesh_terms: D000595:Amino Acid Sequence; D004926:Escherichia coli; D047668:Inteins; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D014176:Protein Biosynthesis; D015202:Protein Engineering; D019154:Protein Splicing; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry*; D060327:Solid-Phase Synthesis Techniques; D013194:Staining and Labeling
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins