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Hydrogen bonding and equilibrium isotope enrichment in histidine-containing proteins

Nature Structural Biology volume 3, pages 522–531 (1996)Cite this article

Abstract

We have measured deuterium/hydrogen fractionation in three histidine-containing proteins, ecHPr, ecHPr mutant S31A, and bsHPr, and in random coil peptides using NMR and mass spectrometry. The amide protons of unstructured peptides exhibit equilibrium enrichment for deuterium, in agreement with previous studies. Enrichment for both protium and deuterium was observed in both HPrs, with f ractionation factors ranging from 0.63 to 1.41. Enrichment for protium was seen in α-helical secondary structure. ‘Strong’ HBs previously identified by mutagenesis and thermodynamic measurements are significantly enriched for protium. Sites of protium enrichment are conserved in a structural context across species lines, though ecHPr and bsHPr share only 30% sequence identity, suggesting that strong HBs are conserved and may play an important role in stabilizing the folded state.

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  1. Department of Biochemistry, Biomolecular Structure Center, Box 357742, University of Washington, Seattle, Washington, 98195-7742, USA

    Peter M. Bowers & Rachel E. Klevit

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Bowers, P., Klevit, R. Hydrogen bonding and equilibrium isotope enrichment in histidine-containing proteins. Nat Struct Mol Biol 3, 522–531 (1996). https://doi.org/10.1038/nsb0696-522

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