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Quantitative measurement of small through-hydrogen-bond and ‘through-space’1H-113Cd and1H-199Hg J couplings in metal-substituted rubredoxin fromPyrococcus furiosus

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Summary

A method is described for measurement of small unresolvable heteronuclear J couplings. The method is based on quantitative analysis of a phase-purged heteronuclear spin-echo difference spectrum, and is demonstrated for measuring1H-113Cd and1H-199Hg J couplings in metal-substituted rubredoxin (Mr ∼ 5.4 kDa) fromPyrococcus furiosus. Couplings from cadmium to backbone amide protons that are hydrogen bonded to the Cys-S atoms directly bonded to Cd vary from smaller than 0.3 to 1.8 Hz; a ‘through-space’ coupling between Cd and the protons of an alanine methyl group was measured to be 0.3 Hz. Couplings to199Hg are significantly larger and fall in the 0.4–4 Hz range.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Maryland Baltimore County, 21228, Baltimore, MD, USA

    Paul R. Blake, Brian Lee & Michael F. Summers

  2. Department of Biochemistry and Center for Metalloenzyme Studies, University of Georgia, 30602, Athens, GA, USA

    Michael W. W. Adams, Jac-Bum Park & Zhi Hao Zhou

  3. Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 20892, Bethesda, MD, USA

    Ad Bax

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  1. Paul R. Blake
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  2. Brian Lee
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  3. Michael F. Summers
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  4. Michael W. W. Adams
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  5. Jac-Bum Park
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  7. Ad Bax
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Blake, P.R., Lee, B., Summers, M.F. et al. Quantitative measurement of small through-hydrogen-bond and ‘through-space’1H-113Cd and1H-199Hg J couplings in metal-substituted rubredoxin fromPyrococcus furiosus . J Biomol NMR 2, 527–533 (1992). https://doi.org/10.1007/BF02192814

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  • DOI: https://doi.org/10.1007/BF02192814

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