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Angiotensin-Responsive Adrenal Glomerulosa Cell Proteins: Characterization by Protease Mapping, Species Comparison, and Specific Angiotensin Receptor Antagonists¹

William S. Middleton Memorial Veterans Hospital and the Departments of Medicine and Pharmacology, University of Wisconsin School of Medicine, Madison, Wisconsin 53705

Address all correspondence and requests for reprints to: Mary E. Elliott, Ph.D., Hypertension Research Laboratory, Room C4114, William S. Middleton Memorial Veterans Hospital, 2500 Overlook Terrace, Madison, Wisconsin 53705.

Angiotensin II (AngII)-stimulated aldosterone synthesis is mediated by the AngII type 1 (AT₁) receptor and requires ongoing protein synthesis. Hormonally-stimulated turnover of a family of 28- to 30-kDa proteins (p30, or steroidogenic acute regulatory proteins) has been linked to enhanced steroid synthesis in several tissues. Our previous work showed that AngII, dibutyryl cAMP, potassium, and atrial natriuretic peptide affected labeling of a group of eight proteins (four of 28 kDa and four of 30 kDa) in bovine adrenal glomerulosa cells. This report extends our findings in three ways: 1) The eight [³⁵S]-methionine-labeled p30 proteins in bovine cells were compared with each other by chymotryptic peptide mapping. Similarity in maps indicated that the eight proteins share a common primary structure. 2) Dibutyryl cAMP treatment of rat adrenal glomerulosa cells affected the levels of four 28-kDa proteins and one 35-kDa protein, whereas AngII affected two of the 28-kDa proteins. There were no responsive 30-kDa proteins in rats comparable with those seen in bovine cells. These results indicate a species difference in the affected proteins. 3) The AT₁ receptor antagonist, losartan, inhibited the effects of AngII on aldosterone synthesis and turnover of the p30 proteins in bovine adrenal glomerulosa cells. PD123319, an antagonist specific for the AngII type 2 receptor, did not block AngII-stimulated aldosterone synthesis and had much less effect on p30 protein labeling than did losartan. These results add to the growing body of evidence that this family of p30 or steroidogenic acute regulatory proteins plays a role in the acute regulation of steroidogenesis by a wide variety of stimulatory hormones in several tissues and species. In addition, losartan’s inhibition of AngII’s effects on the p30 proteins is consistent with a key role for these proteins in processes linking occupation of the AT₁ receptor to stimulation of aldosterone synthesis.

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