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An approach to global fold determination using limited NMR data from larger proteins selectively protonated at specific residue types

Journal of Biomolecular NMR volume 8, pages 360–368 (1996)Cite this article

Summary

A combination of calculation and experiment is used to demonstrate that the global fold of larger proteins can be rapidly determined using limited NMR data. The approach involves a combination of heteronuclear triple resonance NMR experiments with protonation of selected residue types in an otherwise completely deuterated protein. This method of labelling produces proteins with α-specific deuteration in the protonated residues, and the results suggest that this will improve the sensitivity of experiments involving correlation of side-chain (1H and 13C) and backbone (1H and 15N) amide resonances. It will allow the rapid assignment of backbone resonances with high sensitivity and the determination of a reasonable structural model of a protein based on limited NOE restraints, an application that is of increasing importance as data from the large number of genome sequencing projects accumulates. The method that we propose should also be of utility in extending the use of NMR spectroscopy to determine the structures of larger proteins.

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Affiliations

  1. Cambridge Centre for Molecular Recognition, Department of Biochemistry, University of Cambridge, Tennis Court Road, CB2 1QW, Cambridge, UK

    Brian O. Smith, Andrew Raine, Sarah Teichmann, Liat Ben-Tovim, Daniel Nietlispach, R. William Broadhurst & Ernest D. Laue

  2. Laboratory of Cellular and Molecular Biology, Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, 351-01, Wako, Saitama, Japan

    Yutaka Ito & Takehiko Shibata

  3. Cellular Signalling Laboratory, Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, 351-01, Wako, Saitama, Japan

    Tohru Terada & Shigeyuki Yokoyama

  4. Department of Biophysics and Biochemistry, School of Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113, Tokyo, Japan

    Tohru Terada & Shigeyuki Yokoyama

  5. European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69012, Heidelberg, Germany

    Mark Kelly & Hartmut Oschkinat

Authors
  1. Brian O. Smith
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  2. Yutaka Ito
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  3. Andrew Raine
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  4. Sarah Teichmann
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  5. Liat Ben-Tovim
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  6. Daniel Nietlispach
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  7. R. William Broadhurst
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  8. Tohru Terada
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  9. Mark Kelly
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  10. Hartmut Oschkinat
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  11. Takehiko Shibata
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  12. Shigeyuki Yokoyama
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  13. Ernest D. Laue
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Additional information

The first two authors contributed equally to this work.

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Smith, B.O., Ito, Y., Raine, A. et al. An approach to global fold determination using limited NMR data from larger proteins selectively protonated at specific residue types. J Biomol NMR 8, 360–368 (1996). https://doi.org/10.1007/BF00410335

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  • DOI: https://doi.org/10.1007/BF00410335

Keywords

  • Isotope labelling
  • Deuteration
  • Resonance assignment
  • Global fold
  • Larger proteins
  • ras p21