Journal of Biomolecular NMR

, Volume 20, Issue 1, pp 71–75

A method for efficient isotopic labeling of recombinant proteins

  • Authors
  • Jonathan Marley
  • Min Lu
  • Clay Bracken
Article

DOI: 10.1023/A:1011254402785

Cite this article as:
Marley, J., Lu, M. & Bracken, C. J Biomol NMR (2001) 20: 71. doi:10.1023/A:1011254402785

Abstract

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-enrichmentdeuterationEscherichia coliheteronuclear NMRisotope labelingprotein expression

Copyright information

© Kluwer Academic Publishers 2001