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Triple resonance-based 13Cα and 13Cβ CEST experiments for studies of ms timescale dynamics in proteins

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Abstract

A pair of triple resonance based CEST pulse schemes are presented for measuring 13Cα and 13Cβ chemical shifts of sparsely populated and transiently formed conformers that are invisible to traditional NMR experiments. CEST profiles containing dips at resonance positions of 13Cα or 13Cβ spins of major (ground) and minor (excited) conformers are obtained in a pseudo 3rd dimension that is generated by quantifying modulations of cross peaks in 15N, 1HN correlation spectra. An application to the folding reaction of a G48A mutant of the Fyn SH3 domain is presented, illustrating and validating the methodology.

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Acknowledgments

D.L. and A.S. are recipients of post-doctoral scholarships from the Canadian Institutes of Health Research (CHIR). This work 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 Molecular Genetics, Biochemistry and Chemistry, The University of Toronto, Toronto, ON, M5S 1A8, Canada

    Dong Long, Ashok Sekhar & Lewis E. Kay

  2. Program in Molecular Structure and Function, Hospital for Sick Children, 555 University Avenue, Toronto, ON, M5G 1X8, Canada

    Lewis E. Kay

Authors
  1. Dong Long
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  2. Ashok Sekhar
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  3. Lewis E. Kay
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Correspondence to Lewis E. Kay.

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Long, D., Sekhar, A. & Kay, L.E. Triple resonance-based 13Cα and 13Cβ CEST experiments for studies of ms timescale dynamics in proteins. J Biomol NMR 60, 203–208 (2014). https://doi.org/10.1007/s10858-014-9868-5

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  • DOI: https://doi.org/10.1007/s10858-014-9868-5

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