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Demonstration of functionally different interactions between phospholipase C-gamma and the two types of platelet-derived growth factor receptors

Eriksson, Anders; Nånberg, Eeva; Rönnstrand, Lars LU ; Engström, Ulla; Hellman, Ulf; Rupp, Eva; Carpenter, Graham; Heldin, Carl-Henrik and Claesson-Welsh, Lena (1995) In Journal of Biological Chemistry 270(13). p.7773-7781
Abstract
Phosphorylated tyrosine residues in receptor tyrosine kinases serve as binding sites for signal transduction molecules. We have identified two autophosphorylation sites, Tyr-988 and Tyr-1018, in the platelet-derived growth factor (PDGF) alpha-receptor carboxyl-terminal tail, which are involved in binding of phospholipase C-gamma (PLC-gamma). The capacities of the Y988F and Y1018F mutant PDGF alpha-receptors, expressed in porcine aortic endothelial cells, to bind PLC-gamma are 60 and 5% of that of the wild-type receptor, respectively. Phosphorylated but not unphosphorylated peptides containing Tyr-1018 are able to compete with the intact receptor for binding to immobilized PLC-gamma SH2 domains; a phosphorylated Tyr-988 peptide competes 10... (More)
Phosphorylated tyrosine residues in receptor tyrosine kinases serve as binding sites for signal transduction molecules. We have identified two autophosphorylation sites, Tyr-988 and Tyr-1018, in the platelet-derived growth factor (PDGF) alpha-receptor carboxyl-terminal tail, which are involved in binding of phospholipase C-gamma (PLC-gamma). The capacities of the Y988F and Y1018F mutant PDGF alpha-receptors, expressed in porcine aortic endothelial cells, to bind PLC-gamma are 60 and 5% of that of the wild-type receptor, respectively. Phosphorylated but not unphosphorylated peptides containing Tyr-1018 are able to compete with the intact receptor for binding to immobilized PLC-gamma SH2 domains; a phosphorylated Tyr-988 peptide competes 10 times less efficiently. The complex between PLC-gamma and the PDGF alpha-receptor is more stable than that of PLC-gamma and the PDGF beta-receptor. However, PDGF stimulation results in a smaller fraction of tyrosine-phosphorylated PLC-gamma and a smaller accumulation of inositol trisphosphate in cells expressing the alpha-receptor as compared with cells expressing the beta-receptor. We conclude that phosphorylated Tyr-988 and Tyr-1018 in the PDGF alpha-receptor carboxyl-terminal tail bind PLC-gamma, but this association leads to only a relatively low level of tyrosine phosphorylation and activation of PLC-gamma. (Less)
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@article{734992bd-8e52-41c5-9de2-9262322bf9c3,
  abstract     = {Phosphorylated tyrosine residues in receptor tyrosine kinases serve as binding sites for signal transduction molecules. We have identified two autophosphorylation sites, Tyr-988 and Tyr-1018, in the platelet-derived growth factor (PDGF) alpha-receptor carboxyl-terminal tail, which are involved in binding of phospholipase C-gamma (PLC-gamma). The capacities of the Y988F and Y1018F mutant PDGF alpha-receptors, expressed in porcine aortic endothelial cells, to bind PLC-gamma are 60 and 5% of that of the wild-type receptor, respectively. Phosphorylated but not unphosphorylated peptides containing Tyr-1018 are able to compete with the intact receptor for binding to immobilized PLC-gamma SH2 domains; a phosphorylated Tyr-988 peptide competes 10 times less efficiently. The complex between PLC-gamma and the PDGF alpha-receptor is more stable than that of PLC-gamma and the PDGF beta-receptor. However, PDGF stimulation results in a smaller fraction of tyrosine-phosphorylated PLC-gamma and a smaller accumulation of inositol trisphosphate in cells expressing the alpha-receptor as compared with cells expressing the beta-receptor. We conclude that phosphorylated Tyr-988 and Tyr-1018 in the PDGF alpha-receptor carboxyl-terminal tail bind PLC-gamma, but this association leads to only a relatively low level of tyrosine phosphorylation and activation of PLC-gamma.},
  author       = {Eriksson, Anders and Nånberg, Eeva and Rönnstrand, Lars and Engström, Ulla and Hellman, Ulf and Rupp, Eva and Carpenter, Graham and Heldin, Carl-Henrik and Claesson-Welsh, Lena},
  issn         = {1083-351X},
  keyword      = {Platelet-Derived Growth Factor/*metabolism
Recombinant Proteins/pharmacology
Sequence Homology,Platelet-Derived Growth Factor alpha
Receptor,Platelet-Derived Growth Factor beta
Receptors,Site-Directed
Oligodeoxyribonucleotides
Peptide Fragments/chemistry/isolation & purification
Phospholipases/*metabolism
Phosphopeptides/pharmacology
Phosphorylation
Phosphotyrosine
Platelet-Derived Growth Factor/pharmacology
Point Mutation
Receptor Protein-Tyrosine Kinases/metabolism
Receptor,Vascular/*metabolism
Isoenzymes/*metabolism
Kinetics
Molecular Sequence Data
Mutagenesis,Amino Acid Sequence
Animals
Aorta
Base Sequence
Binding,Competitive
Cell Division
Endothelium,Amino Acid
Swine
Thymidine/metabolism
Tyrosine/analogs & derivatives/metabolism},
  language     = {eng},
  number       = {13},
  pages        = {7773--7781},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Demonstration of functionally different interactions between phospholipase C-gamma and the two types of platelet-derived growth factor receptors},
  volume       = {270},
  year         = {1995},
}