title: Measuring hydrogen exchange in proteins by selective water saturation in (1)H- (15)N SOFAST/BEST-type experiments: advantages and limitations.
abstract: HET(ex)-SOFAST NMR (Schanda et al. in J Biomol NMR 33:199-211, 2006) has been proposed some years ago as a fast and sensitive method for semi-quantitative measurement of site-specific amide-water hydrogen exchange effects along the backbone of proteins. Here we extend this concept to BEST readout sequences that provide a better resolution at the expense of some loss in sensitivity. We discuss the theoretical background and implementation of the experiment, and demonstrate its performance for an intrinsically disordered protein, 2 well folded globular proteins, and a transiently populated folding intermediate state. We also provide a critical evaluation of the level of accuracy that can be obtained when extracting quantitative exchange rates from HET(ex) NMR measurements.
authors.affiliation: Institut de Biologie Structurale, Université Grenoble 1, 71 avenue des Martyrs, 38044, Grenoble Cedex 9, France.
authors.forename: Enrico
authors.initials: E
authors.lastname: Rennella
authors.forename: Zsofia
authors.initials: Z
authors.lastname: Solyom
authors.forename: Bernhard
authors.initials: B
authors.lastname: Brutscher
mesh_terms: D000577:Amides / Q000737:chemistry; D057773:Cold Shock Proteins and Peptides / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D006801:Humans; D006859:Hydrogen / Q000737:chemistry*; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D011506:Proteins / Q000737:chemistry*; D066244:Proton Magnetic Resonance Spectroscopy; D025801:Ubiquitin / Q000737:chemistry; D014867:Water / Q000737:chemistry*; D051844:alpha-Synuclein / Q000737:chemistry; D001613:beta 2-Microglobulin / Q000737:chemistry
chemical_list: D000577:Amides; D057773:Cold Shock Proteins and Peptides; C498192:CspA protein, E coli; D029968:Escherichia coli Proteins; D009587:Nitrogen Isotopes; D011506:Proteins; D025801:Ubiquitin; D051844:alpha-Synuclein; D001613:beta 2-Microglobulin; D014867:Water; D006859:Hydrogen