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Anal Biochem. 2012 Jul 15;426(2):126-8. doi: 10.1016/j.ab.2012.04.019. Epub 2012 Apr 23.

One-step amino acid selective isotope labeling of proteins in prototrophic Escherichia coli strains.

Author information

1
Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N 5E5.

Abstract

Amino acid selective isotope labeling is a useful approach to simplification of nuclear magnetic resonance (NMR) spectra of large proteins. Cell-free protein synthesis offers essentially unlimited flexibility of labeling patterns but is labor-intensive and expensive. In vivo labeling is simple in principle but generally requires auxotrophic strains, inhibitors of amino acid synthesis, or complex media formulations. We describe a simple procedure for amino acid selective labeling of proteins expressed in prototrophic Escherichia coli strains. Excellent labeling selectivity was achieved for histidine, lysine, methionine, and alanine. Simplicity and robustness of this protocol make it a useful tool for protein NMR.

PMID:
22538396
DOI:
10.1016/j.ab.2012.04.019
[Indexed for MEDLINE]

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