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The Rac–RhoGDI complex and the structural basis for the regulation of Rho proteins by RhoGDI

Nature Structural Biology volume 7, pages 122–126 (2000)Cite this article

Abstract

Rho family-specific guanine nucleotide dissociation inhibitors (RhoGDIs) decrease the rate of nucleotide dissociation and release Rho proteins such as RhoA, Rac and Cdc42 from membranes, forming tight complexes that shuttle between cytosol and membrane compartments. We have solved the crystal structure of a complex between the RhoGDI homolog LyGDI and GDP-bound Rac2, which are abundant in leukocytes, representing the cytosolic, resting pool of Rho species to be activated by extracellular signals. The N-terminal domain of LyGDI (LyN), which has been reported to be flexible in isolated RhoGDIs, becomes ordered upon complex formation and contributes more than 60% to the interface area. The structure is consistent with the C-terminus of Rac2 binding to a hydrophobic cavity previously proposed as isoprenyl binding site. An inner segment of LyN forms a helical hairpin that contacts mainly the switch regions of Rac2. The architecture of the complex interface suggests a mechanism for the inhibition of guanine nucleotide dissociation that is based on the stabilization of the magnesium (Mg2+) ion in the nucleotide binding pocket.

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Figure 1: Overview of the Rac2–LyGDI complex.
Figure 2: Complex interface in sequence and structure.
Figure 3: Surface representation and hydrophobic cavity.

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Acknowledgements

We thank A. Wittinghofer for initiating the collaboration, for critical comments on the manuscript and encouragement, the EMBL Grenoble outstation, in particular A. Perrakis for providing support for measurements on the microfocus beam line (ID13) at the ESRF under the European Union TMR/LSF Programme, EMBL Heidelberg for preliminary mass spectrometry experiments, B. Prakash and A. Becker for discussions, W. Kabsch for discussion of crystallography, and K. Holmes for continuous support. K.S. thanks the Peter und Traudl Engelhorn Stiftung (Penzberg, Germany) for support in the initial phase of the project. This work was supported in part by grants from the Deutsche Forschungsgemeinschaft and the Medical Faculty of the University of Ulm.

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  1. Klaus Scheffzek

    Present address: European Molecular Biology Laboratory (EMBL), Structural Biology Programme, Meyerhofstr. 1, 69117, Heidelberg, Germany

Authors and Affiliations

  1. Abteilung Strukturelle Biologie, Max-Planck-Institut für molekulare Physiologie, Otto-Hahn-Str. 11, Dortmund, 44227, Germany

    Klaus Scheffzek

  2. Abteilung Biophysik, Max-Planck-Institut für medizinische Forschung, Jahnstr. 29, Heidelberg, 69120, Germany

    Klaus Scheffzek

  3. Abteilung Pharmakologie und Toxikologie Albert-Einstein-Allee 11, Universität Ulm, Ulm, 89081, Germany

    Ilona Stephan, Daria Illenberger & Peter Gierschik

  4. Department of Molecular Biology, Odense University, Odense, 5230, Denmark

    Ole N. Jensen

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Correspondence to Klaus Scheffzek.

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Scheffzek, K., Stephan, I., Jensen, O. et al. The Rac–RhoGDI complex and the structural basis for the regulation of Rho proteins by RhoGDI. Nat Struct Mol Biol 7, 122–126 (2000). https://doi.org/10.1038/72392

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