On the mechanism of action of adrenocorticotropic hormone. The role of ACTH-stimulated phosphorylation and dephosphorylation of adrenal proteins

The regulatory role of phosphorylation of adrenal proteins as it relates to the mechanism of action of adrenocorticotropic hormone (ACTH) has been studied. ACTH, cyclic AMP, or cyclic GMP were added to rat adrenal quarters which had been preincubated with [³²P]phosphate. ³²P-labeled proteins in subcellular fractions were identified after separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The addition of ACTH consistently resulted in the phosphorylation and dephosphorylation of specific adrenal proteins and produced characteristic phosphorylation patterns (autoradiographs) for each subcellular fraction which were very different from control. The changes in phosphorylation of proteins preceded corticosterone production. Also, the degree of phosphorylation of these specific proteins followed a dose-response relationship with ACTH which correlated well to the dose-response for corticosterone production. When cAMP was added to adrenal quarters, the resulting phosphorylation changes were identical to those induced by ACTH. When cGMP was added to adrenal quarters, the resulting phosphorylation patterns were very similar to those produced by control incubations. ACTH or cAMP stimulated corticosterone production 6-fold when compared to control or cGMP-treated tissue. These results suggest the tropic action of ACTH is mediated by cAMP by both phosphorylation and dephosphorylation of specific adrenal proteins.