Journal of Biomolecular NMR

, Volume 20, Issue 1, pp 71-75

First online:

A method for efficient isotopic labeling of recombinant proteins

  • Jonathan Marley
  • , Min Lu
  • , Clay Bracken

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A rapid and efficient approach for preparing isotopically labeled recombinant proteins is presented. The method is demonstrated for 13C labeling of the C-terminal domain of angiopoietin-2, 15N labeling of ubiquitin and for 2H/13C/15N labeling of the Escherichia coli outer-membrane lipoprotein Lpp-56. The production method generates cell mass using unlabeled rich media followed by exchange into a small volume of labeled media at high cell density. Following a short period for growth recovery and unlabeled metabolite clearance, the cells are induced. The expression yields obtained provide a fourfold to eightfold reduction in isotope costs using simple shake flask growths.

13C- 15N-enrichment deuteration Escherichia coli heteronuclear NMR isotope labeling protein expression