A simple method for amino acid selective isotope labeling of recombinant proteins in E. coli

Kit I. Tong, Masayuki Yamamoto, Toshiyuki Tanaka

Research output: Contribution to journalArticlepeer-review

49 Citations (Scopus)

Abstract

A simple and user-friendly method of labeling protein selectively with amino acids in vivo is introduced. This technique does not require the use of transaminase-deficient or auxotrophic strains. By manipulating the product feedback inhibitory loops of the E. coli amino acid metabolic pathways and, if necessary, by using enzyme inhibitors, proteins were labeled efficiently in vivo even with amino acid types that are central to the metabolic pathways, such as glutamine. The sequential backbone resonance assignment of the Neh2 domain of Nrf2 transcriptional factor, an intrinsically disordered protein with high spectral degeneracy, was achieved using this labeling method.

Original languageEnglish
Pages (from-to)59-67
Number of pages9
JournalJournal of biomolecular NMR
Volume42
Issue number1
DOIs
Publication statusPublished - 2008

Keywords

  • Amino acid selective isotope labeling
  • Intrinsically disordered protein
  • N-glutamate labeling
  • N-glutamine labeling
  • Neh2

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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