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J Biomol NMR. 2001 May;20(1):71-5.

A method for efficient isotopic labeling of recombinant proteins.

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  • 1Department of Biochemistry, Weill Medical College of Cornell University, New York, NY 10021, USA.

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.

PMID:
11430757
[PubMed - indexed for MEDLINE]
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