Calmodulin plus cyclic AMP-dependent phosphorylation of a Mr 22,000 pituitary protein.
Protein phosphorylation was examined in cytosolic extracts of adult rat anterior pituitary. In the presence of both cyclic AMP and calmodulin, the phosphorylation of a Mr 22,000 protein was markedly stimulated. Cyclic AMP and calmodulin must both be present in order for this effect to be observed; cyclic GMP does not substitute for cyclic AMP, and the effect is abolished by either trifluoperazine or the heat-stable inhibitor of cyclic AMP-dependent protein kinase. Two-dimensional isoelectric focusing sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicates that there are three molecular species of the Mr 22,000 phosphoprotein, with pI values ranging from 6.8 to 8.1. Phosphorylation of this protein is maximally stimulated by 5 microM cyclic AMP and 5.7 microM calmodulin. The effect of cyclic AMP plus calmodulin is enhanced by preincubation and requires a divalent cation; maximal phosphorylation takes place at 100 microM Mn2+, although higher concentrations of Mg2+ and Co2+ support an equivalent degree of phosphorylation. Cyclic AMP plus calmodulin-dependent protein phosphorylation was not detected in other rat tissues surveyed, including brain, testes, adrenal, kidney, liver, spleen, skeletal muscle, pineal, or posterior pituitary. These results help to explain the previous findings of Brattin and Portanova (Brattin, W.J., Jr., and Portanova, R. (1981) Mol. Cell. Endocr. 23, 77-90) of in vivo but not in vitro phosphorylation of three Mr 20,000 anterior pituitary proteins and indicate a possible point of convergence for calcium and cyclic AMP actions in the anterior pituitary.
D000818:Animals; D002147:Calmodulin / Q000378:metabolism*; D003035:Cobalt / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism*; D004533:Egtazic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008970:Molecular Weight; D010766:Phosphorylation; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D051381:Rats
D002147:Calmodulin; D011506:Proteins; D003035:Cobalt; D008345:Manganese; D004533:Egtazic Acid; D000242:Cyclic AMP; D011494:Protein Kinases; D008274:Magnesium