In Vivo Biosynthesis of a beta-Amino Acid-Containing Protein

Clarissa Melo Czekster, Wesley E. Robertson, Allison S. Walker, Dieter Soell, Alanna Schepartz*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

61 Citations (Scopus)

Abstract

It has recently been reported that ribosomes from erythromycin-resistant Escherichia coli strains, when isolated in S30 extracts and incubated with chemically mis-acylated tRNA, can incorporate certain beta-amino acids into full length DHFR in vitro. Here we report that wild-type E. coli EF-Tu and phenylalanyl-tRNA synthetase collaborate with these mutant ribosomes and others to incorporate beta(3)-Phe analogs into full length DHFR in vivo. E. coli harboring the most active mutant ribosomes are robust, with a doubling time only 14% longer than wild-type. These results reveal the unexpected tolerance of E. coli and its translation machinery to the beta(3)-amino acid backbone and should embolden in vivo selections for orthogonal translational machinery components that, incorporate diverse beta-amino acids into proteins and peptides. E. coli harboring mutant ribosomes may possess the capacity to incorporate many non-natural, non alpha-amino acids into proteins and other sequence programmed polymeric materials.

Original languageEnglish
Pages (from-to)5194-5197
Number of pages4
JournalJournal of the American Chemical Society
Volume138
Issue number16
Early online date18 Apr 2016
DOIs
Publication statusPublished - 27 Apr 2016

Keywords

  • TRANSFER-RNA-SYNTHETASE
  • SITE-SPECIFIC INCORPORATION
  • PEPTIDE-BOND FORMATION
  • GENETIC-CODE
  • EF-TU
  • BACKBONE MUTATIONS
  • MODIFIED RIBOSOMES
  • AMINOACYLATION
  • CHANNEL
  • TRANSLATION

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