Multiple incorporation of non-natural amino acids into a single protein using tRNAs with non-standard structures

Takashi Ohtsuki, Taishi Manabe, Masahiko Sisido

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

The ability to introduce non-natural amino acids into proteins opens up new vistas for the study of protein structure and function. This approach requires suppressor tRNAs that deliver the non-natural amino acid to a ribosome associated with an mRNA containing an expanded codon. The suppressor tRNAs must be absolutely protected from aminoacylation by any of the aminoacyl-tRNA synthetases in the protein synthesizing system, or a natural amino acid will be incorporated instead of the non-natural amino acid. Here, we found that some tRNAs with non-standard structures could work as efficient four-base suppressors fulfilling the above orthogonal conditions. Using these tRNAs, we successfully demonstrated incorporation of three different non-natural amino acids into a single protein.

Original languageEnglish
Pages (from-to)6769-6774
Number of pages6
JournalFEBS Letters
Volume579
Issue number30
DOIs
Publication statusPublished - Dec 19 2005

Keywords

  • Four-base codon
  • Mitochondrial tRNA
  • Non-natural amino acid
  • Orthogonal tRNA
  • tRNA

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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