A small-molecule-based approach to sense codon-templated natural-unnatural hybrid peptides. Selective silencing and reassignment of the sense codon by orthogonal reacylation stalling at the single-codon level.
In the presence of the stable sulfamoyl analogue of phenylalanyl adenylate (Phe-SA), the UUU/UUC sense codon for phenylalanine (Phe) can be silenced and reassigned to a naphthylalanine (Nap) conjugated to tRNAPhe. We have demonstrated the efficiency and selectivity or orthogonality of the Phe-to-Nap reassignment induced by an "orthogonal reacylation stalling" strategy at the single-codon level in the translation of mRNAs of dihydrofolate reductase and a 24-mer oligopeptide. We used a prokaryotic translation system with an essential preincubation, during which the endogenous precharged phenylalanyl-tRNAPhe undergoes deacylation and the reacylation of the resulting tRNAPhe is stalled by the action of Phe-SA to inhibit the phenylalanyl-tRNA synthetase activity. We discuss the significance of the present small-molecule-based approach to sense-codon templated natural-unnatural peptides.
Sando
Shinsuke
S
Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan. ssando@sbchem.kyoto-u.ac.jp
Kanatani
Keiichiro
K
Sato
Nobuhiko
N
Matsumoto
Hironori
H
Hohsaka
Takahiro
T
Aoyama
Yasuhiro
Y
D000215:Acylation; D000241:Adenosine / Q000031:analogs & derivatives; D000409:Alanine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D003062:Codon / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D020868:Gene Silencing; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000096:biosynthesis / Q000138:chemical synthesis* / Q000235:genetics; D010649:Phenylalanine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010652:Phenylalanine-tRNA Ligase / Q000037:antagonists & inhibitors / Q000378:metabolism; D012333:RNA, Messenger / Q000138:chemical synthesis / Q000235:genetics; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis / Q000235:genetics
D003062:Codon; D009842:Oligopeptides; D012333:RNA, Messenger; D012346:RNA, Transfer, Amino Acyl; D010649:Phenylalanine; C059031:1-naphthylalanine; D013762:Tetrahydrofolate Dehydrogenase; D010652:Phenylalanine-tRNA Ligase; D000241:Adenosine; D000409:Alanine