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Crystal structure of a small G protein in complex with the GTPase-activating protein rhoGAP

Nature volume 388, pages 693–697 (1997)Cite this article

Abstract

Small G proteins transduce signals from plasma-membrane receptors to control a wide range of cellular functions1,2. These proteins are clustered into distinct families but all act as molecular switches, active in their GTP-bound form but inactive when GDP-bound. The Rho family of G proteins, which includes Cdc42Hs, activate effectors involved in the regulation of cytoskeleton formation, cell proliferation and the JNK signalling pathway3,4,5,6,7,8,9. G proteins generally have a low intrinsic GTPase hydrolytic activity but there are family-specific groups of GTPase-activating proteins (GAPs) that enhance the rate of GTP hydrolysis by up to 105times10,11. We report here the crystal structure of Cdc42Hs, with the non-hydrolysable GTP analogue GMPPNP, in complex with the GAP domain of p50rhoGAP at 2.7 å resolution. In the complex Cdc42Hs interacts, mainly through its switch I and II regions, with a shallow pocket on rhoGAP which is lined with conserved residues. Arg 85 of rhoGAP interacts with the P-loop of Cdc42Hs, but from biochemical data and by analogy with the G-protein subunit Giα1 (ref. 12), we propose that it adopts a different conformation during the catalytic cycle which enables it to stabilize the transition state of the GTP-hydrolysis reaction.

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Figure 2: Electron density omit map for the bound GMPPNP·Mg.
Figure 1: a, The structure of the Cdc42Hs·GMPPNP/p50rhoGAP complex viewed along the protein–protein interface.
Figure 3: Comparison of the Cdc42Hs·GMPPNP/p50rhoGAP complex with the structure of transducin-α·GTPγS.

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Acknowledgements

We thank M. Hirshberg for making coordinates of Rac1 available before general release, E. Skordalakes for assistance with data collection, A. Hall for the gift of rhoA and rhoGAP expression systems, E. Manser and L. Lim for construction and characterization of the Cdc42HsΔ7 expression system, G. Dodson and D. Trentham for critical reading of this manuscript, and A. Savoia (Trieste) and A. Thompson (ESRF) for beamline assistance. D.O. and E.L. Thank the European commission for financial support and the BBSRC and Welcome Trust for support to the Cambridge Centre for Molecular Recognition. K.R. is funded by an EEC TMR fellowship.

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  1. Katrin Rittinger, Philip A. Walker, Darerca Owen and Ernest Laue: These authors contributed equally to this work.

Authors and Affiliations

  1. National Institute for Medical Research, The Ridgeway, Mill Hill, NW7 1AA, London, UK

    Katrin Rittinger, Philip A. Walker, John F. Eccleston, Kurshid Nurmahomed, Steven J. Gamblin & Stephen J. Smerdon

  2. Department of Biochemistry, University of Cambridge, Tennis Court Road, CB2 1QW, Cambridge, UK

    Darerca Owen & Ernest Laue

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Correspondence to Steven J. Gamblin.

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Rittinger, K., Walker, P., Eccleston, J. et al. Crystal structure of a small G protein in complex with the GTPase-activating protein rhoGAP. Nature 388, 693–697 (1997). https://doi.org/10.1038/41805

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