A rapid and robust method for selective isotope labeling of proteins

Myat T. Lin, Lindsay J. Sperling, Heather L. Frericks Schmidt, Ming Tang, Rimma I. Samoilova, Takashi Kumasaka, Toshio Iwasaki, Sergei A. Dikanov, Chad Rienstra, Robert B. Gennis

Research output: Contribution to journalArticlepeer-review


Amino-acid selective isotope labeling of proteins offers numerous advantages in mechanistic studies by revealing structural and functional information unattainable from a crystallographic approach. However, efficient labeling of proteins with selected amino acids necessitates auxotrophic hosts, which are often not available. We have constructed a set of auxotrophs in a commonly used Escherichia coli expression strain C43(DE3), a derivative of E. coli BL21(DE3), which can be used for isotopic labeling of individual amino acids or sets of amino acids. These strains have general applicability to either soluble or membrane proteins that can be expressed in E. coli. We present examples in which proteins are selectively labeled with 13C- and 15N-amino acids and studied using magic-angle spinning solid-state NMR and pulsed EPR, demonstrating the utility of these strains for biophysical characterization of membrane proteins, radical-generating enzymes and metalloproteins.

Original languageEnglish (US)
Pages (from-to)370-378
Number of pages9
Issue number4
StatePublished - Dec 2011


  • Auxotroph
  • EPR
  • Escherichia coli
  • Isotope
  • NMR
  • Protein expression

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)


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