The kinetics of steroidogenesis activator polypeptide in the rat adrenal cortex. Effects of adrenocorticotropin, cyclic adenosine 3':5'-monophosphate, cycloheximide, and circadian rhythm.
The behavior of steroidogenesis activator polypeptide (SAP), a recently described modulator of cholesterol side-chain cleavage activity (Pedersen, R. C., and Brownie, A. C. (1987) Science 236, 188-190), was investigated in rat adrenocortical cells using a specific radioimmunoassay. In response to a maximal dose of adrenocorticotropic hormone (ACTH) (1 nM) or of 8-Br-cAMP (1 mM), an increase in intracellular SAP begins rapidly (less than 1 min) and reaches half-maximal and maximal levels (16-fold greater than basal) at 3 and 15 min, respectively. A plateau at this maximal concentration of SAP is then maintained. The levels of intracellular SAP content and of corticosterone output exhibit a similar dose-dependent response to ACTH (EC50 = 25 and 30 pM, respectively). Treatment of ACTH-stimulated cells with cycloheximide reverses the rise in SAP (t1/2 congruent to 5-7 min). In vivo the SAP content of adrenals from quiescent rats is concordant with the circadian rhythm of the pituitary-adrenal axis; at the apex (1800 h), adrenal SAP is 13-fold higher than at the nadir (0800 h), paralleling 2- and 7-fold variations in cholesterol side-chain cleavage activity and serum corticosterone levels, respectively. At both time points, SAP levels rise in response to stress. Of the rat tissues examined, only the major steroid-forming organs (adrenal cortex and gonads) had significant levels of immunoreactive, cAMP-responsive SAP, while cAMP-unresponsive immunoreactivity was also detectable in the thymus, spleen, and brain. Considered together with the biological activity previously demonstrated for SAP in vitro, these data are consistent with its role as a cAMP-dependent, cycloheximide-sensitive modulator of steroid biosynthesis.
Mertz
L M
LM
Department of Biochemistry, School of Medicine and Biomedical Sciences, State University of New York, Buffalo 14214.
Pedersen
R C
RC
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology*; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D002940:Circadian Rhythm; D003345:Corticosterone / Q000096:biosynthesis / Q000097:blood; D000242:Cyclic AMP / Q000502:physiology*; D003513:Cycloheximide / Q000494:pharmacology*; D005260:Female; D006360:Heat-Shock Proteins; D007700:Kinetics; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000378:metabolism*; D014176:Protein Biosynthesis; D011506:Proteins / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values; D000091342:Endoplasmic Reticulum Chaperone BiP
D015124:8-Bromo Cyclic Adenosine Monophosphate; D000324:Adrenocorticotropic Hormone; D003994:Bucladesine; D002786:Cholesterol Side-Chain Cleavage Enzyme; D003345:Corticosterone; D000242:Cyclic AMP; D003513:Cycloheximide; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D011506:Proteins; C051684:GRP78 protein, rat; D000091342:Endoplasmic Reticulum Chaperone BiP