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A single-quantum methyl 13C-relaxation dispersion experiment with improved sensitivity

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Abstract

A pulse sequence is described for recording single-quantum 13C-methyl relaxation dispersion profiles of 13C-selectively labeled methyl groups in proteins that offers significant improvements in sensitivity relative to existing approaches where initial magnetization derives from 13C polarization. Sensitivity gains in the new experiment are achieved by making use of polarization from 1H spins and 1H → 13C → 1H type magnetization transfers. Its utility has been established by applications involving three different protein systems ranging in molecular weight from 8 to 28 kDa, produced using a number of different selective labeling approaches. In all cases exchange parameters from both 13C→1H and 1H → 13C → 1H classes of experiment are in good agreement, with gains in sensitivity of between 1.7 and 4-fold realized using the new scheme.

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Acknowledgments

P. L. and P. V. are supported by fellowships from the Hellmuth Hertz foundation and the Canadian Institutes of Health Research (CIHR) Training Grant in Protein Folding and Disease. This research was supported by a grant from the CIHR. L.E.K. holds a Canada Research Chair in Biochemistry.

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Authors and Affiliations

  1. Departments of Medical Genetics, Biochemistry and Chemistry, The University of Toronto, Toronto, ON, Canada, M5S 1A8

    Patrik Lundström, Pramodh Vallurupalli & Lewis E. Kay

  2. Medical Research Council Centre for Protein Engineering, Hills Road, Cambridge, CB2 2QH, UK

    Tomasz L. Religa

  3. Department of Chemistry and Biochemistry, University of California at Santa Barbara, Santa Barbara, CA, 93106-9510, USA

    Frederick W. Dahlquist

Authors
  1. Patrik Lundström
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  2. Pramodh Vallurupalli
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  3. Tomasz L. Religa
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  4. Frederick W. Dahlquist
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  5. Lewis E. Kay
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Correspondence to Lewis E. Kay.

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Lundström, P., Vallurupalli, P., Religa, T.L. et al. A single-quantum methyl 13C-relaxation dispersion experiment with improved sensitivity. J Biomol NMR 38, 79–88 (2007). https://doi.org/10.1007/s10858-007-9149-7

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  • DOI: https://doi.org/10.1007/s10858-007-9149-7

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