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The Role of the Conserved Switch II Glutamate in Guanine Nucleotide Exchange Factor-Mediated Nucleotide Exchange of GTP-Binding Proteins

Gasper, Raphael, Thomas, Christoph, Reza Ahmadian, Mohammad, Wittinghofer, Alfred·2008·Journal of Molecular Biology
DOI 10.1016/j.jmb.2008.03.011
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Abstract

Guanine nucleotide exchange factors (GEFs) regulate the activity of small G proteins by catalysing the intrinsically slow exchange of GDP for GTP. The mechanism involves the formation of trimeric G protein-nucleotide-GEF complexes, followed by the release of nucleotide to form stable binary G protein-GEF complexes. A number of structural studies of G protein-GEF complexes have shown large structural changes induced in the nucleotide binding site. Together with a recent structure of a trimeric complex, these studies have suggested not only some common principles but also large differences in detail in the GEF-mediated exchange reaction. Several structures suggested that a glutamic acid residue in switch II, which is part of the DxxGQE motif and highly conserved in Ras-like G proteins, might have a decisive mechanistic role in GEF-mediated nucleotide exchange reactions. Here we show that mutation of the switch II glutamate to Ala severely impairs GEF-catalysed nucleotide exchange in most, but not all, Ras family G proteins, explaining its high sequence conservation. The residue determines the initial approach of GEF to the nucleotide-loaded G protein and does not appreciably affect the formation of a binary nucleotide-free complex. Its major effect thus appears to be the removal of the P-loop lysine from its interaction with the nucleotide.

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Metadata

Authors
Gasper, R., Thomas, C., Reza Ahmadian, M., & Wittinghofer, A.
Venue
Journal of Molecular Biology
Publisher
Elsevier BV
Published
2008-05
Volume
379
Issue
1
Pages
51-63
ISSN
0022-2836
DOI
10.1016/j.jmb.2008.03.011
Cited by
24
References
1
Language
en
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CoChat. (2026). Citation check: The Role of the Conserved Switch II Glutamate in Guanine Nucleotide Exchange Factor-Mediated Nucleotide Exchange of GTP-Binding Proteins. https://app.cochat.ai/papers/10.1016/j.jmb.2008.03.011
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Gasper, R., Thomas, C., Reza Ahmadian, M., & Wittinghofer, A. (2008). The Role of the Conserved Switch II Glutamate in Guanine Nucleotide Exchange Factor-Mediated Nucleotide Exchange of GTP-Binding Proteins. Journal of Molecular Biology. https://doi.org/10.1016/j.jmb.2008.03.011