title: Selectivity of CDC25 homology domain-containing guanine nucleotide exchange factors.
abstract: The Ras family of small G-proteins plays an essential role in the regulation of a variety of signal transduction processes, ranging from cell cycle control to the regulation of exocytosis. Signalling by the Ras G-proteins is initiated by the CDC25 homology domain (CDC25-HD) containing guanine nucleotide exchange factors (GEFs); each GEF, with its specific selectivity profile towards G-proteins, commonly acts on only a small subset of the Ras family members. Thus, GEFs play a pivotal part in establishing the activation of the downstream signalling routes. The structural basis for the establishment of selectivity in the GEF-G-protein interaction is only partially understood, and several controversies on the selectivity of GEFs are discussed in the literature. In the present study, we undertook a systematic approach to determine the selectivity of CDC25-HD for members of the Ras family. We generated a data set of 126 pairs using a standardised in vitro approach encompassing purified recombinant proteins, and a comprehensive mutational study analysed the basis of the selectivity. Together, these data highlight the distinct selectivity of various GEFs and allow for predictions of untested combinations of GEFs and G-proteins.
authors: Molecular Cancer Research, Centre of Biomedical Genetics and Cancer Genomics Centre, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: Milica
authors: M
authors: Popovic
authors: Marije
authors: M
authors: Rensen-de Leeuw
authors: Holger
authors: H
authors: Rehmann
mesh_terms: D000595:Amino Acid Sequence; D004252:DNA Mutational Analysis; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction; D013379:Substrate Specificity; D020666:ras-GRF1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D020666:ras-GRF1; D019204:GTP-Binding Proteins
keywords: 2′-/3′-O-(N-methylanthraniloyl)-guanosine diphosphate; CDC25 homology domain; CDC25-HD; GAP; GEF; GST; GTPase-activating protein; Ras family; glutathione S-transferase; guanine nucleotide exchange factor; mGDP; selectivity; small G-protein