Processing of calcitonin and epidermal growth factor after binding to receptors in human breast cancer cells (T 47D).
125I-labelled calcitonin and 125I-labelled epidermal growth factor (EGF) bound to T 47D breast cancer cells at 37 degrees C in a manner that became increasingly resistant to removal by acid pH. Bound 125I-labelled EGF became resistant to acid removal more rapidly than did bound 125I-labelled calcitonin. The shift from acid accessibility to acid inaccessibility was energy-dependent since it was not seen at 4 degrees C and was inhibited in the presence of cell metabolic inhibitors. Radioactivity removed by acid represented intact hormone as assessed by trichloroacetic acid precipitation, whereas radioactivity released spontaneously by the cells was trichloroacetic acid-soluble. Inclusion of 10 mM-NH4Cl in the incubation medium resulted in an accumulation of cell-associated radioactivity without affecting the shift to acid inaccessibility. The accumulated radioactivity was relatively more trichloroacetic acid-precipitable in comparison with that associated with control cells. These data are consistent with internalization of receptor-bound EGF and a similar though slower mechanism of processing for receptor-bound calcitonin. The predominant route of hormone release from cells seems to occur via intracellular degradation rather than dissociation from cell-surface receptors.
D000643:Ammonium Chloride / Q000494:pharmacology; D000963:Antimetabolites / Q000494:pharmacology; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002116:Calcitonin / Q000378:metabolism*; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018003:Receptors, Calcitonin; D011956:Receptors, Cell Surface / Q000378:metabolism*
D000963:Antimetabolites; D018003:Receptors, Calcitonin; D011956:Receptors, Cell Surface; D000643:Ammonium Chloride; D004815:Epidermal Growth Factor; D002116:Calcitonin; D066246:ErbB Receptors