Journal of Biomolecular NMR

, 42:35 | Cite as

Measurement of carbonyl chemical shifts of excited protein states by relaxation dispersion NMR spectroscopy: comparison between uniformly and selectively 13C labeled samples

  • Patrik Lundström
  • D. Flemming Hansen
  • Lewis E. Kay


Carr–Purcell–Meiboom–Gill (CPMG) relaxation dispersion nuclear magnetic resonance (NMR) spectroscopy has emerged as a powerful method for quantifying chemical shifts of excited protein states. For many applications of the technique that involve the measurement of relaxation rates of carbon magnetization it is necessary to prepare samples with isolated 13C spins so that experiments do not suffer from magnetization transfer between coupled carbon spins that would otherwise occur during the CPMG pulse train. In the case of 13CO experiments however the large separation between 13CO and 13Cα chemical shifts offers hope that robust 13CO dispersion profiles can be recorded on uniformly 13C labeled samples, leading to the extraction of accurate 13CO chemical shifts of the invisible, excited state. Here we compare such chemical shifts recorded on samples that are selectively labeled, prepared using [1-13C]-pyruvate and NaH13CO3, or uniformly labeled, generated from 13C-glucose. Very similar 13CO chemical shifts are obtained from analysis of CPMG experiments recorded on both samples, and comparison with chemical shifts measured using a second approach establishes that the shifts measured from relaxation dispersion are very accurate.


Relaxation dispersion NMR Chemical exchange Carr–Purcell–Meiboom–Gill Excited protein states Chemical shifts 



Nuclear magnetic resonance





We thank Dr. Elliott Stollar and Ms. Hong Lin for the gift of Ark1p peptide that was used in some of the experiments. This work was supported by a grant from the Canadian Institutes of Health Research (CIHR). P. L. and D. F. H. hold fellowships from the CIHR Training Grant on Protein Folding in Health and Disease (P. L.) and the CIHR (D. F. H.). The authors thank Dr. Pramodh Vallurupalli for useful discussions. L. E. K. is the recipient of a Canada Research Chair in Biochemistry.


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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Patrik Lundström
    • 1
    • 2
    • 3
  • D. Flemming Hansen
    • 1
    • 2
    • 3
  • Lewis E. Kay
    • 1
    • 2
    • 3
  1. 1.Department of Medical GeneticsThe University of TorontoTorontoCanada
  2. 2.Department of BiochemistryThe University of TorontoTorontoCanada
  3. 3.Department of ChemistryThe University of TorontoTorontoCanada

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