title: An atypical naturally split intein engineered for highly efficient protein labeling.
abstract: Protein trans-splicing catalyzed by split inteins is a powerful technique for assembling a polypeptide backbone from two separate parts. However, split inteins with robust efficiencies and short fragments suitable for peptide synthesis are rare and have mostly been artificially created. The novel split intein AceL-TerL was identified from metagenomic data and characterized. It represents the first naturally occurring, atypically split intein. The N-terminal fragment of only 25 amino acids is the shortest natural intein fragment to date and was easily amenable to chemical synthesis with a fluorescent label. Optimal protein trans-splicing activity was observed at low temperatures. Further improved mutants were selected by directed protein evolution. The engineered intein variants with up to 50-fold increased rates showed unprecedented efficiency in chemically labeling of a diverse set of proteins. These inteins should prove valuable tools for protein semi-synthesis and other intein-related biotechnological applications.
authors.affiliation: Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Münster, Wilhelm-Klemm-Strasse 2, 48149 Münster (Germany).
authors.forename: Ilka V
authors.initials: IV
authors.lastname: Thiel
authors.forename: Gerrit
authors.initials: G
authors.lastname: Volkmann
authors.forename: Shmuel
authors.initials: S
authors.lastname: Pietrokovski
authors.forename: Henning D
authors.initials: HD
authors.lastname: Mootz
mesh_terms: D000595:Amino Acid Sequence; D002384:Catalysis; D005452:Fluoresceins / Q000737:chemistry; D047668:Inteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015202:Protein Engineering; D019154:Protein Splicing; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D005452:Fluoresceins; D011994:Recombinant Proteins; C024098:6-carboxyfluorescein; D013879:Thioredoxins
keywords: autocatalysis; chemoenzymatic synthesis; combinatorial mutagenesis; protein engineering; protein semisynthesis