title: Kinetic analysis of the leucyl/phenylalanyl-tRNA-protein transferase with acceptor peptides possessing different N-terminal penultimate residues.
abstract: The introduction of non-natural amino acids at the N-terminus of peptides/proteins using leucyl/phenylalanyl-tRNA-protein transferase (L/F-transferase) is a useful technique for protein engineering. To accelerate the chemoenzymatic reaction, here we systematically optimized the N-terminal penultimate residue of the acceptor peptide. Positively charged, small, or hydrophilic amino acids at this position show positive effects for the reaction. Kinetic analysis of peptides possessing different penultimate residues suggests that the side chain of the residue affects peptide-binding affinity towards the L/F-transferase. These findings also provide biological insight into the effect of the penultimate amino acid on substrate specificity of natural proteins to be degraded via the N-end rule pathway.
authors.affiliation: Department of Engineering Science, Bioscience and Technology Program, The Graduate School of Informatics and Engineering, The University of Electro-Communications (UEC), 7-5-1 Chofugaoka, Chofu, Tokyo 182-8585, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Kawaguchi
authors.forename: Kumino
authors.initials: K
authors.lastname: Maejima
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kuroiwa
authors.forename: Masumi
authors.initials: M
authors.lastname: Taki
keywords: Enzyme kinetics; L/F-transferase; N-terminal penultimate residue; Non-natural amino acid (unnatural amino acid); Protein engineering; The N-end rule
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