Characterization of cyclic AMP-binding proteins in rat sertoli cells using a photoaffinity ligand.
Protein-bound cyclic AMP (cAMP) levels in cultured rat Sertoli cells have been determined after exposure to follicle-stimulating hormone (FSH) and agents which elevate intracellular cAMP or mimic cAMP action. Changes in the content of protein-bound cAMP were correlated with changes in receptor availability determined by measuring [3H] cAMP binding. Using the photoaffinity analog of cAMP, 8-N3 [32P] cAMP, two major cAMP-binding proteins in Sertoli cell cytosol, with molecular weights of 47 000 and 53 000 daltons, were identified as regulatory subunits of type I and type II cAMP-dependent protein kinases, respectively. Densitometric analysis of autoradiograms demonstrated differential activation of the two isozymes in response to treatment with FSH and other agents. Results of this study demonstrate the value of measuring changes in protein-bound cAMP and the utility of the photoaffinity labeling technique in correlating hormone-dependent processes in which activation of cAMP-dependent protein kinase occurs.
D000345:Affinity Labels; D000818:Animals; D001386:Azides; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000031:analogs & derivatives; D002373:Cyclic AMP Receptor Protein; D004789:Enzyme Activation / Q000187:drug effects; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D008297:Male; D008970:Molecular Weight; D051381:Rats; D011953:Receptors, Cyclic AMP / Q000378:metabolism; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism
D000345:Affinity Labels; D001386:Azides; D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D011953:Receptors, Cyclic AMP; C017042:8-azidoadenosine-3',5'-monophosphate; D005640:Follicle Stimulating Hormone; D000242:Cyclic AMP