PMID:14709072

Citation	Patching, SG, Herbert, RB, O'Reilly, J, Brough, AR and Henderson, PJ (2004) Low 13C-background for NMR-based studies of ligand binding using 13C-depleted glucose as carbon source for microbial growth: 13C-labeled glucose and 13C-forskolin binding to the galactose-H+ symport protein GalP in Escherichia coli. J. Am. Chem. Soc. 126:86-7
Abstract	Obtrusive 13C-backgrounds can be a problem in 13C NMR-based studies of ligand binding to bacterial membrane transport proteins in their natural state in inner membranes. This is largely solved for the bacterial galactose-H+ symport protein GalP by growing the producing organism Escherichia coli on 13C-depleted glucose (13C </= 0.07%) as the main carbon source. 13C solid-state NMR-based binding studies for the inhibitor forskolin 1 and the transported substrate glucose 2, both singly labeled with 13C, are reported and discussed. For 1, tight binding is observed, while for 2, significant exchange takes place during the time scale of the NMR experiment.
Links	PubMed Online version:10.1021/ja038275c
Keywords	Calcium-Binding Proteins; Carbon Isotopes; Escherichia coli/metabolism; Forskolin/metabolism; Glucose/chemistry; Glucose/metabolism; Isotope Labeling; Ligands; Monosaccharide Transport Proteins/metabolism; Nuclear Magnetic Resonance, Biomolecular/methods; Periplasmic Binding Proteins/metabolism; Protein Binding
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