Binding and internalization of epidermal growth factor by rat pituitary tumor cells.
Epidermal growth factor (EGF) regulates hormone synthesis and decreases the growth rate of clonal lines of rat pituitary tumor cells. [125I]iodo-EGF bound to specific receptors on GH4C1 rat pituitary cells with an apparent Kd of 0.34 nM. At 0 degrees C, 84% of specifically bound [125I]iodo-EGF appeared to be on the cell surface, while at 37 degrees C, 84% of bound [125I]iodo-EGF appeared to be internalized and could not be removed by treatment with an acidic high-salt buffer. Internalization of surface-bound [125I]iodo-EGF required 10 min to reach completion and was markedly temperature-dependent. Monodansylcadaverine, bacitracin, ammonium chloride, chloroquine, quinacrine and vinblastin did not inhibit internalization of [125I]iodo-EGF, suggesting that transglutaminase is not involved; but these drugs did cause an increased accumulation of radioactivity, probably by inhibiting degradation. In 2 h at 37 degrees C, approx. 35% of cell-associated [125I]iodo-EGF was degraded as analyzed by gel filtration, and 72% of the material that dissociated from the cells migrated as low molecular weight material or iodo-Tyr. Incubation of GH4C1 cells with unlabeled EGF caused a prolonged and dose-related decrease in the ability of the cells to bind [125I]iodo-EGF in a subsequent incubation, i.e. down-regulation. The pathway for internalization and degradation of the EGF-receptor complex in GH4C1 cells resembles that described for fibroblasts, in which the cellular response is different, but differs from the behavior of other hormone-receptor complexes in GH4C1 pituitary cells.
D000818:Animals; D002103:Cadaverine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors; D010911:Pituitary Neoplasms / Q000378:metabolism*; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
D011956:Receptors, Cell Surface; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; C008542:monodansylcadaverine; D002103:Cadaverine