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

J Am Chem Soc. 2005 Jun 8;127(22):7998-9. doi: 10.1021/ja0502977.

Abstract

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.

MeSH terms

  • Acylation
  • Adenosine / analogs & derivatives
  • Alanine / analogs & derivatives*
  • Alanine / chemistry
  • Alanine / genetics
  • Amino Acid Sequence
  • Codon / chemistry*
  • Codon / genetics
  • Codon / metabolism
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Gene Silencing
  • Molecular Sequence Data
  • Oligopeptides / biosynthesis
  • Oligopeptides / chemical synthesis*
  • Oligopeptides / genetics
  • Phenylalanine / chemistry
  • Phenylalanine / genetics
  • Phenylalanine / metabolism
  • Phenylalanine-tRNA Ligase / antagonists & inhibitors
  • Phenylalanine-tRNA Ligase / metabolism
  • RNA, Messenger / chemical synthesis
  • RNA, Messenger / genetics
  • RNA, Transfer, Amino Acyl / chemistry*
  • RNA, Transfer, Amino Acyl / genetics
  • RNA, Transfer, Amino Acyl / metabolism
  • Tetrahydrofolate Dehydrogenase / biosynthesis
  • Tetrahydrofolate Dehydrogenase / genetics

Substances

  • Codon
  • Oligopeptides
  • RNA, Messenger
  • RNA, Transfer, Amino Acyl
  • Phenylalanine
  • 1-naphthylalanine
  • Tetrahydrofolate Dehydrogenase
  • Phenylalanine-tRNA Ligase
  • Adenosine
  • Alanine