Calcium and protein kinase C enhance parathyroid hormone- and forskolin-stimulated adenylate cyclase in ROS 17/2.8 cells.
Both parathyroid hormone (PTH)- and forskolin-stimulated adenylate cyclase activities in ROS 17/2.8 cells are enhanced by increasing the medium concentrations of CaCl2 from 10(-5) M to 3 x 10(-3) M. The ED50 for CaCl2 for both PTH- and forskolin-stimulated activities are similar. The tumor-promoting phorbol ester phorbol 12-myristate 13-acetate (PMA), a known activator of protein kinase C, also enhanced both PTH- and forskolin-stimulated adenylate cyclase. This action of PMA is specific for protein kinase C as phorbol esters that are not activators of protein kinase C had no effect on the system. The combined effects of PMA and CaCl2 were more than additive. The separate and combined effects of PMA and CaCl2 changed the rate of activation of the enzyme (Vmax) but did not modify the ED50 for PTH or for forskolin. PMA and CaCl2 both enhanced the potentiating effect of submaximal dose of forskolin on PTH-stimulated adenylate cyclase. It is concluded that calcium and PMA enhance PTH-sensitive adenylate cyclase and increase the production of cAMP by a mechanism that appears to involve the catalytic subunit of the enzyme and probably its interaction with a guanine nucleotide regulatory protein.
Rao
L G
LG
Division of Endocrinology and Metabolism, St. Michael's Hospital, Toronto, Ontario, Canada.
Murray
T M
TM
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology*; D012516:Osteosarcoma / Q000201:enzymology / Q000473:pathology*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D011493:Protein Kinase C / Q000494:pharmacology*; D051381:Rats; D015290:Second Messenger Systems / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000201:enzymology / Q000473:pathology
D010281:Parathyroid Hormone; D005576:Colforsin; D011493:Protein Kinase C; D000262:Adenylyl Cyclases; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium