Journal of Biomolecular NMR

, Volume 50, Issue 1, pp 13–18 | Cite as

Measuring 1HN temperature coefficients in invisible protein states by relaxation dispersion NMR spectroscopy

  • Guillaume Bouvignies
  • Pramodh Vallurupalli
  • Matthew H. J. Cordes
  • D. Flemming Hansen
  • Lewis E. Kay
Communication

Abstract

A method based on the Carr-Purcell-Meiboom-Gill relaxation dispersion experiment is presented for measuring the temperature coefficients of amide proton chemical shifts of low populated ‘invisible’ protein states that exchange with a ‘visible’ ground state on the millisecond time-scale. The utility of the approach is demonstrated with an application to an I58D mutant of the Pfl6 Cro protein that undergoes exchange between the native, folded state and a cold denatured, unfolded conformational ensemble that is populated at a level of 6% at 2.5°C. A wide distribution of amide temperature coefficients is measured for the unfolded state. The distribution is centered about –5.6 ppb/K, consistent with an absence of intra-molecular hydrogen bonds, on average. However, the large range of values (standard deviation of 2.1 ppb/K) strongly supports the notion that the unfolded state of the protein is not a true random coil polypeptide chain.

Keywords

CPMG relaxation dispersion Temperature coefficients Amide protons Cold denaturation 

Supplementary material

10858_2011_9498_MOESM1_ESM.docx (783 kb)
Supplementary material 1 (DOCX 783 kb)

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Guillaume Bouvignies
    • 1
  • Pramodh Vallurupalli
    • 1
  • Matthew H. J. Cordes
    • 2
  • D. Flemming Hansen
    • 1
    • 3
  • Lewis E. Kay
    • 1
  1. 1.Departments of Molecular Genetics, Biochemistry and ChemistryThe University of TorontoTorontoCanada
  2. 2.Department of Chemistry and BiochemistryThe University of ArizonaTucsonUSA
  3. 3.Institute of Structural and Molecular BiologyUniversity College LondonLondonUK

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