title: Exploration of incorporation of Nalpha-methylated amino acids into peptides by sense-suppression method.
abstract: Here we describe the ribosomal synthesis of peptides containing an N(alpha)-methylated amino acid by using sense-suppression method. To incorporate the various N(alpha)-methylated amino acids into the peptide, we utilized recently developed two technologies, a ribozyme-based tRNA acylation system (Flexizyme system) and a reconstituted cell-free translation system (PURE system). Firstly, various N(alpha)-methylated amino acids were charged onto a suppressor tRNA by Flexizyme system. Then, using PURE system, we constructed a vacant codon corresponding to the suppressed position. By combining these systems, we were able to examine various N(alpha)-methylated amino acids for the single incorporation into the peptide backbone and determined those with high efficiencies. Thus, this demonstration opens a new door to develop a new methodology for the synthesis of peptides containing multiple N-methylated backbone via the genetic code reprogramming.
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo,7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kawakami
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murakami
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Suga
mesh_terms: D002474:Cell-Free System; D003062:Codon; D008745:Methylation; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry*; D016337:RNA, Catalytic / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D013489:Suppression, Genetic
chemical_list: D003062:Codon; D010455:Peptides; D016337:RNA, Catalytic; D012343:RNA, Transfer