Platelet-derived growth factor receptor-beta (PDGFR-beta) activation promotes its association with the low density lipoprotein receptor-related protein (LRP). Evidence for co-receptor function

J Biol Chem. 2005 Jul 29;280(30):27872-8. doi: 10.1074/jbc.M505410200. Epub 2005 Jun 8.

Abstract

Activation of the platelet-derived growth factor receptor-beta (PDGFR-beta) leads to tyrosine phosphorylation of the cytoplasmic domain of LRP and alters its association with adaptor and signaling proteins, such as Shc. The mechanism of the PDGF-induced LRP tyrosine phosphorylation is not well understood, especially since PDGF not only activates PDGF receptor but also binds directly to LRP. To gain insight into this mechanism, we used a chimeric receptor in which the ligand binding domain of the PDGFR-beta was replaced with that from the macrophage colony-stimulating factor (M-CSF) receptor, a highly related receptor tyrosine kinase of the same subfamily, but with different ligand specificity. Activation of the chimeric receptor upon the addition of M-CSF readily mediated the tyrosine phosphorylation of LRP. Since M-CSF is not recognized by LRP, these results indicated that growth factor binding to LRP is not necessary for this phosphorylation event. Using a panel of cytoplasmic domain mutants of the chimeric M-CSF/PDGFR-beta, we confirmed that the kinase domain of PDGFR-beta is absolutely required for LRP tyrosine phosphorylation but that PDGFR-beta-mediated activation of phosphatidylinositol 3-kinase, RasGAP, SHP-2, phospholipase C-gamma, and Src are not necessary for LRP tyrosine phosphorylation. To identify the cellular compartment where LRP and the PDGFR-beta may interact, we employed immunofluorescence and immunogold electron microscopy. In WI-38 fibroblasts, these two receptors co-localized in coated pits and endosomal compartments following PDGF stimulation. Further, phosphorylated forms of the PDGFR-beta co-immunoprecipitated with LRP following PDGF treatment. Together, these studies revealed close association between activated PDGFR-beta and LRP, suggesting that LRP functions as a co-receptor capable of modulating the signal transduction pathways initiated by the PDGF receptor from endosomes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cytoplasm / metabolism
  • Endosomes / metabolism
  • Fibroblasts / metabolism
  • Humans
  • Immunohistochemistry
  • Immunoprecipitation
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Ligands
  • Low Density Lipoprotein Receptor-Related Protein-1 / metabolism*
  • Mice
  • Microscopy, Fluorescence
  • Mutation
  • NIH 3T3 Cells
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phospholipase C gamma
  • Phosphorylation
  • Protein Binding
  • Protein Structure, Tertiary
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatases / metabolism
  • Receptor, Macrophage Colony-Stimulating Factor / metabolism
  • Receptor, Platelet-Derived Growth Factor beta / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction
  • Time Factors
  • Type C Phospholipases / metabolism
  • Tyrosine / chemistry
  • Tyrosine / metabolism
  • ras GTPase-Activating Proteins / metabolism

Substances

  • Intracellular Signaling Peptides and Proteins
  • Ligands
  • Low Density Lipoprotein Receptor-Related Protein-1
  • Recombinant Fusion Proteins
  • ras GTPase-Activating Proteins
  • Tyrosine
  • Phosphatidylinositol 3-Kinases
  • Receptor, Macrophage Colony-Stimulating Factor
  • Receptor, Platelet-Derived Growth Factor beta
  • PTPN11 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Protein Tyrosine Phosphatases
  • Ptpn11 protein, mouse
  • Type C Phospholipases
  • Phospholipase C gamma