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A set of HNCO-based experiments for measurement of residual dipolar couplings in 15N, 13C, (2H)-labeled proteins

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Abstract

Several HNCO-based three-dimensional experiments are described for the measurement of 13C′(i−1)-13Cα(i−1), 15N(i)-13C′(i−1), 15N(i)-13Cα(i), 15N(i)-13Cα(i−1), 1HN(i)-13Cα(i), 1HN(i)-13Cα(i−1), and 13Cα(i−1)-13Cβ(i−1) scalar and dipolar couplings in 15N, 13C, (2H)-labelled protein samples. These pulse sequences produce spin-state edited spectra superficially resembling an HNCO correlation spectrum, allowing accurate and simple measurement of couplings without introducing additional spectral crowding. Scalar and dipolar couplings are measured with good sensitivity from relatively large proteins, as demonstrated with three proteins: cardiac Troponin C, calerythrin and ubiquitin. Measurement of several dipolar couplings between spin-1/2 nuclei using spin-state selective 3D HNCO spectra provides a wealth of structural information.

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Authors and Affiliations

  1. Institute of Biotechnology, University of Helsinki, P.O. Box 56, FIN-00014, Helsinki, Finland

    Perttu Permi

  2. Department of Molecular Genetics, Biochemistry, and Microbiology, University of Cincinnati, USA

    Paul R. Rosevear

  3. VTT Biotechnology, FIN-02044 VTT, Espoo, Finland

    Arto Annila

Authors
  1. Perttu Permi
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  2. Paul R. Rosevear
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  3. Arto Annila
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Permi, P., Rosevear, P.R. & Annila, A. A set of HNCO-based experiments for measurement of residual dipolar couplings in 15N, 13C, (2H)-labeled proteins. J Biomol NMR 17, 43–54 (2000). https://doi.org/10.1023/A:1008372624615

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