title: Chemical-tag labeling of proteins using fully recombinant split inteins.
abstract: Chemical-tag labeling of proteins involving split inteins is an approach for the selective chemical modification of proteins without the requirement of any chemical synthesis to be performed. In a two-step protocol, a very short tag fused to a split intein auxiliary protein is first labeled in a bioconjugation reaction with a synthetic moiety either at its N-terminus (amine-tag) or at the side chain of an unnatural amino acid (click-tag). The labeled protein is then mixed with the protein of interest fused to the complementary intein fragment. In the resulting spontaneous protein trans-splicing reaction the split intein fragments remove themselves and ligate the tag to the protein of interest in a virtually traceless fashion. The reaction can be performed either using a purified protein of interest or to label a protein in the context of a living cell. All protein components are recombinantly expressed and all chemical reagents are commercially available.
authors.affiliation: Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.
authors.forename: Anne-Lena
authors.initials: AL
authors.lastname: Bachmann
authors.forename: Julian C J
authors.initials: JC
authors.lastname: Matern
authors.forename: Vivien
authors.initials: V
authors.lastname: Schütz
authors.forename: Henning D
authors.initials: HD
authors.lastname: Mootz
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D057930:Click Chemistry; D004926:Escherichia coli; D047668:Inteins; D051379:Mice; D015202:Protein Engineering; D019154:Protein Splicing; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D013194:Staining and Labeling
chemical_list: D011993:Recombinant Fusion Proteins