Abstract
Activation of the beta-receptor for platelet-derived growth factor (PDGF) by its ligand leads to autophosphorylation on a number of tyrosine residues. Here we show that Tyr763 in the kinase insert region is a novel autophosphorylation site, which after phosphorylation binds the protein tyrosine phosphatase SHP-2. SHP-2 has also previously been shown to bind to phosphorylated Tyr1009 in the PDGF beta-receptor. Porcine aortic endothelial (PAE) cells transfected with a PDGF beta-receptor in which Tyr763 and Tyr1009 were mutated to phenylalanine residues failed to associate with SHP-2 after ligand stimulation. Moreover, PDGF-BB-induced Ras GTP-loading and Erk2 activation were severely compromised in the receptor mutant. Whereas the mitogenic response to PDGF-BB remained at the same level as in cells expressing wild-type PDGF beta-receptor, chemotaxis induced by PDGF-BB was significantly decreased in the case of the Y763F/Y1009F mutant cells, suggesting an important role for SHP-2 in chemotactic signaling.
MeSH terms
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Amino Acid Sequence
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Animals
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Becaplermin
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Binding Sites
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Calcium-Calmodulin-Dependent Protein Kinases / metabolism
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Calcium-Calmodulin-Dependent Protein Kinases / physiology*
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Cells, Cultured
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Chemotaxis*
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Endothelium, Vascular / metabolism
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Enzyme Activation
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Guanosine Triphosphate / metabolism
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Intracellular Signaling Peptides and Proteins
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Mice
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Mitogen-Activated Protein Kinase 1
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Phosphorylation
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Phosphotyrosine / metabolism
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Platelet-Derived Growth Factor / pharmacology
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Protein Processing, Post-Translational*
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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Protein Tyrosine Phosphatase, Non-Receptor Type 6
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Protein Tyrosine Phosphatases / metabolism*
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Proto-Oncogene Proteins c-sis
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Receptor, Platelet-Derived Growth Factor beta
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Receptors, Platelet-Derived Growth Factor / chemistry
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Receptors, Platelet-Derived Growth Factor / metabolism*
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Signal Transduction*
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Swine
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Transfection
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Tyrosine / metabolism*
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ras Proteins / physiology*
Substances
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Intracellular Signaling Peptides and Proteins
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Platelet-Derived Growth Factor
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Proto-Oncogene Proteins c-sis
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Becaplermin
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Phosphotyrosine
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Tyrosine
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Guanosine Triphosphate
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Receptor, Platelet-Derived Growth Factor beta
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Receptors, Platelet-Derived Growth Factor
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Calcium-Calmodulin-Dependent Protein Kinases
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Mitogen-Activated Protein Kinase 1
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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Protein Tyrosine Phosphatase, Non-Receptor Type 6
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Protein Tyrosine Phosphatases
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Ptpn11 protein, mouse
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Ptpn6 protein, mouse
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ras Proteins