title: Effect of adrenocorticotropin on steroid 21-hydroxylase synthesis and activity in cultured bovine adrenocortical cells. Increased synthesis in the absence of increased activity.
abstract: The action of adrenocorticotropin (ACTH) to stimulate synthesis of steroid 21-hydroxylase was studied in bovine adrenocortical cells maintained in primary culture. Continuous treatment with ACTH (1 microM) caused an increased incorporation of [35S]methionine into cytochrome P-450C21 (21-hydroxylase cytochrome P-450); a maximum value (15-fold increase) was attained 24 h after initiation of ACTH treatment. Also, a 3-fold increase in cytochrome P-450C21 synthesis was observed in a cell-free translation system programmed by RNA isolated from cells that were exposed to ACTH for 24 h. The rate of synthesis of cytochrome P-450C21 declined after longer periods of treatment of the cells with ACTH. The increase in synthesis of cytochrome P-450C21 was associated with an increase (3-6 fold) in both total cytochrome P-450 content and in the type I absorbance change induced by 17 alpha-hydroxyprogesterone in microsomes prepared from ACTH-treated cells, as compared with that in microsomes from control cells. By contrast, ACTH did not act to increase steroid 21-hydroxylase activity in cultured intact cells, as determined by the rate of secretion of cortisol and 11-deoxycortisol, after addition of 17 alpha-hydroxyprogesterone to the medium. Similarly, there was no difference in steroid 21-hydroxylase activity in postmitochondrial supernatant fractions prepared from non-treated or ACTH-treated cells. Cytochrome P-450C21 was found to be synthesized as a form identical in molecular weight to the mature form. These results are indicative that ACTH acts to stimulate the synthesis of steroid 21-hydroxylase, yet is without a demonstrable effect on the activity of this enzyme which is high throughout the time period of the experiment.
mesh_terms: D000302:Adrenal Cortex / Q000187:drug effects / Q000201:enzymology*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D002417:Cattle; D002474:Cell-Free System; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis; D004790:Enzyme Induction; D013255:Steroid 21-Hydroxylase / Q000096:biosynthesis*; D013250:Steroid Hydroxylases / Q000096:biosynthesis*
chemical_list: D000324:Adrenocorticotropic Hormone; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D013255:Steroid 21-Hydroxylase