Measuring hydrogen exchange in proteins by selective water saturation in (1)H- (15)N SOFAST/BEST-type experiments: advantages and limitations

J Biomol NMR. 2014 Nov;60(2-3):99-107. doi: 10.1007/s10858-014-9857-8. Epub 2014 Aug 31.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemistry
  • Cold Shock Proteins and Peptides / chemistry
  • Escherichia coli Proteins / chemistry
  • Humans
  • Hydrogen / chemistry*
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular*
  • Proteins / chemistry*
  • Proton Magnetic Resonance Spectroscopy*
  • Ubiquitin / chemistry
  • Water / chemistry*
  • alpha-Synuclein / chemistry
  • beta 2-Microglobulin / chemistry


  • Amides
  • Cold Shock Proteins and Peptides
  • CspA protein, E coli
  • Escherichia coli Proteins
  • Nitrogen Isotopes
  • Proteins
  • Ubiquitin
  • alpha-Synuclein
  • beta 2-Microglobulin
  • Water
  • Hydrogen