epidermal-growth-factor and 8-azidoadenosine-3--5--monophosphate

The activity of the two cAMP-dependent protein kinases (PKa I and PKa II) was evaluated in dog thyroid cells in primary cultures after a 6-day growth period induced by either thyrotropin (TSH) or epidermal growth factor (EGF). Although the total PKa activity was not affected in cells cultured in the presence of TSH or EGF, their actions on the PKa I and PKa II expressions were significantly different. The activity of PKa I was strongly inhibited by TSH (70-80%) while with EGF it was either stimulated or unaffected with respect to controls. The two mitogens did not have a significant effect on the activity of PKa II. Forskolin (Fk) mimicked the effect of TSH. The expression of the two regulatory subunits (R I and R II), evaluated by the covalent binding of 8-azido-cAMP, was similar to the expression of the corresponding catalytic activities, suggesting a coregulation of the catalytic and regulatory subunits from the same isozyme. After chronic stimulation by TSH, differentiated dog thyroid cells are almost completely deprived of PKa I.

Topics: Affinity Labels; Animals; Azides; Cells, Cultured; Chromatography, DEAE-Cellulose; Colforsin; Cyclic AMP; Cytosol; Dogs; Electrophoresis; Epidermal Growth Factor; Isoenzymes; Kinetics; Protein Kinase Inhibitors; Thyroid Gland; Thyrotropin