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Rapid Characterization of Hydrogen Exchange in Proteins


  • H.S.A. acknowledges research support from DAE, DST, and DBT research grants. P.D.S. acknowledges support from Wellcome Trust International Senior Research Fellowship in Biomedical Science (UK), WT081643MA. We thank Dr. John Cort, PNNL, for providing the ubiquitin plasmid and Garima Jaipuria, IISc, for help in recording NMR spectra.


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Kinetics and thermodynamics of amide hydrogen exchange in proteins can be investigated with two-dimensional 13CO–15N NMR correlation experiments (see picture). The spectra are acquired with high resolution and sensitivity. A single type of experiment on one sample serves to characterize hydrogen–deuterium fractionation factors and hydrogen-exchange rates that span three orders of magnitude.