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Pressure-induced local unfolding of the Ras binding domain of RalGDS

Nature Structural Biology volume 7, pages 547–550 (2000)Cite this article

Abstract

The reliable prediction of the precise three-dimensional structure of proteins from their amino acid sequence is a major, still unresolved problem in biochemistry. Pressure is a parameter that controls folding/unfolding transitions of proteins through the volume change ΔV of the protein-solvent system. By varying the pressure from 30 to 2,000 bar we detected using 15N/1H 2D NMR spectroscopy a unique equilibrium unfolding intermediate I in the Ras binding domain of the Ral guanine nucleotide dissociation stimulator (Ral GDS). It is characterized by a local melting of specific structural elements near hydrophobic cavities while the overall folded structure is maintained.

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Figure 1: Pressure effects observable in 15N,1H-HSQC spectra14 of RalGDS-RBD (residues 11–97).
Figure 2: Pressure induced local unfolding of RalGDS-RBD.
Figure 3: Conformational equilibrium of RalGDS-RBD.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft, the European Union, the Humboldt foundation and the Ministry of Education, Science, Sports and Culture of Japan. We thank R. Jaenicke and A. Wittinghofer for helpful discussions, and M. Geyer for aid with the sample preparation.

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Authors and Affiliations

  1. Department of Molecular Science, Graduate School of Science and Technology, Kobe University, Kobe, 657-8501 , Japan

    Kyoko Inoue & Kazuyuki Akasaka

  2. Faculty of Science, Kobe University, Kobe , 657-8501, Japan

    Hiroaki Yamada & Kazuyuki Akasaka

  3. Department of Structural Biology, Max-Planck-Institute for Molecular Physiology, Dortmund, 44227, Germany

    Christian Herrmann

  4. Institute of Biophysics and Physical Biochemistry, University of Regensburg, Regensburg, 93040, Germany

    Werner Kremer, Till Maurer, Rolf Döker & Hans Robert Kalbitzer

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  1. Kyoko Inoue
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Correspondence to Kazuyuki Akasaka or Hans Robert Kalbitzer.

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Inoue, K., Yamada, H., Akasaka, K. et al. Pressure-induced local unfolding of the Ras binding domain of RalGDS . Nat Struct Mol Biol 7, 547–550 (2000). https://doi.org/10.1038/76764

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