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A molecular switch between alternative conformational states in the complex of Ran and importin β1

Nature Structural & Molecular Biology volume 10, pages 553–557 (2003)Cite this article

Abstract

Several million macromolecules are exchanged each minute between the nucleus and cytoplasm by receptor-mediated transport. Most of this traffic is controlled by the small GTPase Ran, which regulates assembly and disassembly of the receptor–cargo complexes in the appropriate cellular compartment. Here we applied dynamic force spectroscopy to study the interaction of Ran with the nuclear import receptor importin β1 (impβ) at the single-molecule level. We found that the complex alternates between two distinct conformational states of different adhesion strength. The application of an external mechanical force shifts equilibrium toward one of these states by decreasing the height of the interstate activation energy barrier. The other state can be stabilized by a functional Ran mutant that increases this barrier. These results support a model whereby functional control of Ran–impβ is achieved by a population shift between pre-existing alternative conformations.

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Figure 1: Measurement of single-molecule unbinding forces.
Figure 2: Molecular unbinding of different Ran–impβ complexes.
Figure 3: The Ran–GTP–impβ interaction.

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Acknowledgements

We thank C. Riener and H. Gruber for the synthesis of the PEG spacers, O. Bogin for his help in the expression of impβ and RanQ69L, and D. Görlich for providing an expression construct for impβ. We also thank J. Klafter, A. Filippov, A. Minsky, D. Tawfik, M. Urbakh and E.J. Wachtel for helpful discussions and comments. This work was supported by the Charles H. Revson Foundation (Israel Science Foundation) and the Austrian National Science Foundations.

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Authors and Affiliations

  1. Department of Biological Chemistry, the Weizmann Institute of Science, Rehovot, 76100, Israel

    Reinat Nevo, David Kaftan & Ziv Reich

  2. Institute for Biophysics, University of Linz, Altenbergerstrasse 69, Linz, A-4040, Austria

    Cordula Stroh, Ferry Kienberger & Peter Hinterdorfer

  3. Department of Plant Sciences, Weizmann Institute of Science, Rehovot, 76100, Israel

    Vlad Brumfeld

  4. Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot, 76100, Israel

    Michael Elbaum

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Correspondence to Ziv Reich.

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Nevo, R., Stroh, C., Kienberger, F. et al. A molecular switch between alternative conformational states in the complex of Ran and importin β1. Nat Struct Mol Biol 10, 553–557 (2003). https://doi.org/10.1038/nsb940

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