title: One-step amino acid selective isotope labeling of proteins in prototrophic Escherichia coli strains.
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.
authors.affiliation: Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N 5E5.
authors.forename: Christopher
authors.initials: C
authors.lastname: O'Grady
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Rempel
authors.forename: Akosiererem
authors.initials: A
authors.lastname: Sokaribo
authors.forename: Sergiy
authors.initials: S
authors.lastname: Nokhrin
authors.forename: Oleg Y
authors.initials: OY
authors.lastname: Dmitriev
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism*; D007553:Isotope Labeling; D019906:Nuclear Magnetic Resonance, Biomolecular; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D011506:Proteins