Discovery of aminoacyl-tRNA synthetase activity through cell-surface display of noncanonical amino acids

A. James Link, Mandy K.S. Vink, Nicholas J. Agard, Jennifer A. Prescher, Carolyn R. Bertozzi, David A. Tirrell

Research output: Contribution to journalArticle

133 Scopus citations

Abstract

The incorporation of noncanonical amino acids into recombinant proteins in Escherichia coli can be facilitated by the introduction of new aminoacyl-tRNA synthetase activity into the expression host. We describe here a screening procedure for the identification of new aminoacyl-tRNA synthetase activity based on the cell surface display of noncanonical amino acids. Screening of a saturation mutagenesis library of the E. coli methionyl-tRNA synthetase (MetRS) led to the discovery of three MetRS mutants capable of incorporating the long-chain amino acid azidonorleucine into recombinant proteins with modest efficiency. The Leu-13 → Gly (L13G) mutation is found in each of the three MetRS mutants, and the MetRS variant containing this single mutation is highly efficient in producing recombinant proteins that contain azidonorleucine.

Original languageEnglish (US)
Pages (from-to)10180-10185
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number27
DOIs
StatePublished - Jul 4 2006
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • Azide-alkyne ligation
  • Azidohomoalanine
  • Azidonorleucine
  • Click chemistry

Fingerprint Dive into the research topics of 'Discovery of aminoacyl-tRNA synthetase activity through cell-surface display of noncanonical amino acids'. Together they form a unique fingerprint.

  • Cite this