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Structures of Cdc42 bound to the active and catalytically compromised forms of Cdc42GAP

Nature Structural Biology volume 5, pages 1047–1052 (1998)Cite this article

Abstract

The Rho-related small GTP-binding protein Cdc42 has a low intrinsic GTPase activity that is significantly enhanced by its specific GTPase-activating protein, Cdc42GAP. In this report, we present the tertiary structure for the aluminum fluoride-promoted complex between Cdc42 and a catalytically active domain of Cdc42GAP as well as the complex between Cdc42 and the catalytically compromised Cdc42GAP(R305A) mutant. These structures, which mimic the transition state for the GTP hydrolytic reaction, show the presence of an AlF 3 molecule, as was seen for the corresponding Ras–p120RasGAP complex, but in contrast to what has been reported for the Rho–Cdc42GAP complex or for heterotrimeric G protein α subunits, where AlF 4 – was observed. The Cdc42GAP stabilizes both the switch I and switch II domains of Cdc42 and contributes a highly conserved arginine (Arg 305) to the active site. Comparison of the structures for the wild type and mutant Cdc42GAP complexes provides important insights into the GAP-catalyzed GTP hydrolytic reaction.

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Figure 1: Views of the complex between Cdc42 and the wild type Cdc42GAP.
Figure 2: Stereo view of an omit map for the complex between Cdc42 and the wild type Cdc42GAP, in which GDP, Mg2+ and AlF3, as well as atoms within 3.5 Å, were removed.
Figure 3: Comparison of the complexes between Cdc42 and the wild type Cdc42GAP and the Cdc42GAP(R305A) mutant.
Figure 4: Stereo superposition of the Cdc42-AIF-Cdc42GAP structure and the Giα1-AIF4–-RGS4 complex19.

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Acknowledgements

We thank S. Doublie for invaluable help and comments on this project. We thank R. Sweet for beamtime on X12C and the CHESS staff for help during data collection. We thank J. Stamos for excellent technical assistance, W. Wang for help and C. Westmiller for expert secretarial assistance. This work was supported by grants from the National Institutes of Health and the Human Frontiers Science Program.

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Authors and Affiliations

  1. Department of Pharmacology, College of Veterinary Medicine, Cornell University, Ithaca, 14853, New York, USA

    Nicolas Nassar, Gregory R. Hoffman, Danny Manor & Richard A. Cerione

  2. Department of Chemistry, Baker's Laboratory, Cornell University, Ithaca, 14853, New York, USA

    Nicolas Nassar & Jon C. Clardy

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  1. Nicolas Nassar
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  2. Gregory R. Hoffman
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Correspondence to Richard A. Cerione.

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Nassar, N., Hoffman, G., Manor, D. et al. Structures of Cdc42 bound to the active and catalytically compromised forms of Cdc42GAP. Nat Struct Mol Biol 5, 1047–1052 (1998). https://doi.org/10.1038/4156

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