title: One-pot synthesis of azoline-containing peptides in a cell-free translation system integrated with a posttranslational cyclodehydratase.
abstract: Azoline moieties in the backbones of peptidic natural products are important structural motifs that contribute to diverse bioactivities. Some azoline-containing peptides (Az-peptides) are produced from ribosomally synthesized precursor peptides, in which cysteine, serine, and threonine residues are converted to their corresponding azolines by posttranslational modification through a cyclodehydratase. We have devised an in vitro biosynthesis system of Az-peptides, referred to as the FIT-PatD (flexible in vitro translation) system, by the integration of a cell-free translation system with the posttranslational cyclodehydratase PatD. This system enabled the "one-pot" synthesis of a wide variety of Az-peptide derivatives expressed from synthetic DNA templates. The FIT-PatD system also facilitated mutagenesis studies on a wide array of precursor peptide sequences, unveiling unique in vitro substrate tolerance of PatD.
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan; JST, PRESTO, Tokyo 113-0033, Japan. Electronic address: y-goto@chem.s.u-tokyo.ac.jp.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Goto
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Tokyo 113-8656, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan.
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Kato
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan.
authors.forename: Shotaro
authors.initials: S
authors.lastname: Tsunoda
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan; JST, CREST, Tokyo 113-0033, Japan. Electronic address: hsuga@chem.s.u-tokyo.ac.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Suga
mesh_terms: D002474:Cell-Free System; D006836:Hydro-Lyases / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D053778:Thiazolidines / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010455:Peptides; D053778:Thiazolidines; D006836:Hydro-Lyases