Platelet-derived growth factor receptor inducibility is acquired immediately after translation and does not require glycosylation.
Antibodies to phosphotyrosine were used in immunoprecipitation experiments to determine if post-translational modification of the platelet-derived growth factor (PDGF) receptor was required for the acquisition of ligand-induced tyrosine kinase activity. In intact Balb/c 3T3 fibroblasts, only the fully processed 180-kDa receptor was activated (tyrosine-phosphorylated) by PDGF. In a cell-free assay, however, the tyrosine-phosphorylated forms of the 160- and 145-kDa PDGF receptor precursors were also detected. These activated precursors were immunoprecipitated after brief (5-15 min) metabolic labeling periods. Thus the receptor could bind PDGF and induce tyrosine kinase activity shortly after translation. Unlike the mature form of the receptor, the 160-kDa receptor precursor was resistant to digestion with endo-alpha-N-acetylgalactosaminidase and thus did not contain O-linked oligosaccharides. Since this receptor precursor was activated by PDGF in the cell-free assay, the addition of O-linked sugars must not be necessary for ligand binding activity. Incubation of cells with tunicamycin completely inhibited N-linked glycosylation of the PDGF receptor. Nevertheless, PDGF still induced tyrosine phosphorylation of the 130-kDa aglycoreceptor in lysates of tunicamycin-treated cells. Thus, the addition of N-linked oligosaccharides was also not required for receptor activation. These findings show that the PDGF receptor acquired the ability to be activated by ligand cotranslationally or immediately after translation and that the addition of N- or O-linked oligosaccharides was not required for ligand binding and tyrosine kinase activities.
Keating
M T
MT
Department of Medicine, University of California, San Francisco 94143.
Harryman
C C
CC
Williams
L T
LT
D000818:Animals; D002236:Carbohydrate Conformation; D002460:Cell Line; D006031:Glycosylation; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010982:Platelet-Derived Growth Factor / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000378:metabolism; D017479:Receptors, Platelet-Derived Growth Factor
D010982:Platelet-Derived Growth Factor; D011498:Protein Precursors; D011956:Receptors, Cell Surface; D011505:Protein-Tyrosine Kinases; D017479:Receptors, Platelet-Derived Growth Factor