Down regulation of epidermal growth factor receptors: direct demonstration of receptor degradation in human fibroblasts.
The metabolism of the receptor for epidermal growth factor (EGF) has been measured by labeling the receptor in vivo with radioactive amino acid precursors and then determining, by immunoprecipitation with specific anti-EGF receptor antisera, the rate of degradation of the receptor when the cells are placed in a nonradioactive medium. In human fibroblasts the rate of EGF receptor degradation (t1/2 = 10.1 h) was faster than the rate of degradation of total cell protein. When EGF was added to the nonradioactive medium, the half-life of prelabeled receptor was decreased to 1.2 h in human fibroblasts. These data demonstrate by direct analysis of receptor protein that during "down regulation" the EGF receptor is rapidly degraded. Enhanced receptor degradation was observed 5-10 min after the addition of EGF. The EGF-induced degradation of the receptor was blocked by methylamine, chloroquine, iodoacetate, or incubation at 25 degrees C. We have also shown that EGF-induced down regulation in human fibroblasts results in a decrease in the total amount of EGF receptor protein present. The amount of EGF receptor protein has been quantitated by radiolabeling cellular protein and immunoprecipitation of the receptor. The EGF receptor constitutes approximately 0.0035% of the cellular protein in human fibroblasts.
D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D004705:Endocytosis; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008247:Lysosomes / Q000187:drug effects; D008744:Methylamines / Q000494:pharmacology; D008985:Monensin / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000378:metabolism*
D008744:Methylamines; D011956:Receptors, Cell Surface; D004815:Epidermal Growth Factor; D002738:Chloroquine; D008985:Monensin; D066246:ErbB Receptors