Melittin inhibits epidermal growth factor-induced protein tyrosine phosphorylation: comparison with phorbol myristate acetate and calcium ionophore A23187.
In HER14 cells, epidermal growth factor (EGF) induces tyrosine phosphorylation of several proteins, including its own receptor. The bee venom peptide, melittin, impaired EGF-dependent protein tyrosine phosphorylation in a calcium-dependent manner. The melittin effect was similarly reproduced by calcium ionophore A23187. The effect of melittin and calcium ionophore A23187 on EGF-dependent protein tyrosine phosphorylation was abolished by treatment of cells with the calcium chelator EGTA. Phorbol-myristate acetate (PMA) inhibited EGF-dependent protein tyrosine phosphorylation, and when compared to melittin or calcium ionophore A23187, only PMA potentiated the EGF-induced tyrosine phosphorylation of two proteins immunologically related to mitogen activated protein (MAP) kinases of 40 kDa and 44 kDa molecular mass. Unlike PMA, the effect of melittin and calcium ionophore A23187 on inhibition of EGF-dependent protein tyrosine phosphorylation was lost neither in protein kinase C-depleted cells nor in cells treated with the protein tyrosine phosphatase inhibitors NaF and Na3VO4. Melittin inhibited high affinity binding of EGF to its receptor in intact cells, but this effect was not prevented by EGTA. It is concluded that melittin and calcium ionophore A23187 differ from PMA in their inhibition of EGF-dependent protein tyrosine phosphorylation in vivo, by acting via a Ca(2+)-dependent pathway, that is independent of protein kinase C, protein tyrosine phosphatase activity and high affinity binding of EGF to its receptor.
Department of Pharmacology, New York University Medical Center, NY 10016.
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D016475:3T3 Cells; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D004121:Dimethyl Sulfoxide; D004815:Epidermal Growth Factor / Q000037:antagonists & inhibitors* / Q000378:metabolism / Q000494:pharmacology; D008555:Melitten / Q000494:pharmacology*; D051379:Mice; D010766:Phosphorylation; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014443:Tyrosine / Q000378:metabolism*
D008555:Melitten; D000001:Calcimycin; D014443:Tyrosine; D004815:Epidermal Growth Factor; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium; D004121:Dimethyl Sulfoxide