Fluoride complexes of oncogenic Ras mutants to study the Ras-RasGAP interaction
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Lothar Gremer
Abstract
Down-regulation of Ras signalling is mediated by specific GTPase-activating proteins (GAPs), which stimulate the very slow GTPase reaction of Ras by 105-fold. The basic features of the GAP activity involve the stabilisation of both switch regions of Ras in the transition state, and the insertion of an arginine finger. In the case of oncogenic Ras mutations, the features of the active site are disturbed. To understand these features in more detail, we investigated the effects of oncogenic mutations of Ras and compared the GAP-stimulated GTPase reaction with the ability to form GAP-mediated aluminium or beryllium fluoride complexes. In general, we found a correlation between the size of the amino acid at position 12, the GTPase activity and ability to form aluminium fluoride complexes. While Gly12 is very sensitive to even the smallest possible structural change, Gly13 is much less sensitive to steric hindrance, but is sensitive to charge. Oncogenic mutants of Ras defective in the GTPase activity can however form ground-state GppNHp complexes with GAP, which can be mimicked by beryllium fluoride binding. We show that beryllium fluoride complexes are less sensitive to structural changes and report on a state close to but different from the ground state of the GAP-stimulated GTPase reaction.
©2008 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Review
- Some assembly required: dedicated chaperones in eukaryotic proteasome biogenesis
- Protein Structure and Function
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- Fluoride complexes of oncogenic Ras mutants to study the Ras-RasGAP interaction
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- Molecular mechanisms of aromatase inhibition by new A, D-ring modified steroids
- A common binding motif for various bacteria of the bacteria-binding peptide SRCRP2 of DMBT1/gp-340/salivary agglutinin
- Putative identification of an amphipathic α-helical sequence in hemolysin of Escherichia coli (HlyA) involved in transmembrane pore formation
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- Homologous and heterologous expression and maturation processing of extracellular glutamyl endopeptidase of Staphylococcus epidermidis
- Cleaved SLPI, a novel biomarker of chymase activity
- Expression of tissue and plasma kallikreins and kinin B1 and B2 receptors in lung cancer
- Mechanism of methaemoglobin breakdown by the lysine-specific gingipain of the periodontal pathogen Porphyromonas gingivalis
- Novel Techniques
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