Measurement of intrinsic exchange rates of amide protons in a 15N-labeled peptide.
We have used a modified version of a previously proposed technique, MEXICO [Gemmecker et al. (1993) J. Am. Chem. Soc., 115, 11620], and improved data analysis procedures in order to measure rapid hydrogen exchange (HX) rates of amide protons in peptides labeled only with 15N. The requirement of 13C-/15N-labeled material has been circumvented by adjusting conditions so that NOE effects associated with amide protons can be neglected (i.e., omega 0 tau c approximately 1). The technique was applied to an unstructured 15N-labeled 12-residue peptide to measure intrinsic HX rates, which are the essential reference for examining protein and peptide structure and dynamics through deceleration of HX rates. The method provided accurate HX rates from 0.5 to 50 s-1 under the conditions used. The measured rates were in good agreement with those predicted using correction factors determined by Englander and co-workers [Bai et al. (1993) Proteins, 17, 75], with the largest deviations from the predicted rates found for residues close to the N-terminus. The exchange rates were found to exhibit significant sensitivity to the concentration of salt in the sample.
Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037, USA.
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D000577:Amides / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D009587:Nitrogen Isotopes; D010455:Peptides / Q000737:chemistry*; D011522:Protons
D000577:Amides; D009587:Nitrogen Isotopes; D010455:Peptides; D011522:Protons