Effects of the indolocarbazole 3744W on the tyrosine kinase activity of the cytoplasmic domain of the platelet-derived growth factor beta-receptor.
The cytoplasmic domain of the platelet-derived growth factor (PDGF) beta-receptor was expressed in insect cells by using a baculovirus system. The resulting protein was a constitutively active tyrosine kinase that could phosphorylate both protein and peptide substrates. A recently identified potent and selective inhibitor of intact PDGF receptor autophosphorylation, 3744W, inhibited the autophosphorylation of the cytoplasmic domain both in vitro (IC50 1.8+/-0.12 microM) and within intact insect cells (IC50 2.0 microM). However, under identical assay conditions, 3744W did not inhibit the phosphorylation of the synthetic polymeric peptide poly(Glu4Tyr1) even at concentrations as high as 100 microM. These results suggest that, although 3744W inhibits PDGF receptor autophosphorylation directly, it can discriminate between phosphate acceptor substrates.
Cell Signalling Group, Biology Division, Wellcome Research Laboratories, Beckenham, Kent, UK. iju44964@glaxowellcome.co.uk
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D000818:Animals; D015153:Blotting, Western; D002227:Carbazoles / Q000494:pharmacology*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression; D007303:Insect Vectors; D008954:Models, Biological; D010766:Phosphorylation; D010797:Phthalimides / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism*; D011994:Recombinant Proteins; D013997:Time Factors
C110877:3744 W; D002227:Carbazoles; D004791:Enzyme Inhibitors; D010797:Phthalimides; D011994:Recombinant Proteins; D011505:Protein-Tyrosine Kinases; D017479:Receptors, Platelet-Derived Growth Factor