title: Protein phosphorylation mediates effects of isoproterenol on adenylate cyclase activity in rat cortical membranes.
abstract: The effects of (-)-isoproterenol on adenylate cyclase activity were studied in rat cerebral cortical membranes prepared and assayed in the presence of calcium ions. In assays carried out in the presence of high Mg2+ concentrations (5-10 mM) and of Ca2+ in the micromolar range, addition of 1-100 micro M (-)-isoproterenol caused over 50% inhibition of adenylate cyclase activity. Since these conditions are optimal for supporting endogenous phosphorylative activity in synaptic membranes, we tested whether the observed effects are mediated by changes in the phosphorylation of specific proteins in these membranes. This was done by preincubation of lysed synaptosomes under phosphorylating conditions in the presence and absence of isoproterenol followed by extensive washes and analysis of cyclic AMP formation in resuspended membranes. Addition of (-)-isoproterenol to the preincubation resulted in a 30% decrease of adenylate cyclase activity in the reincubation. Inclusion of [gamma-32P]ATP in the preincubation and examination of the phosphorylation state of specific proteins in membranes entering the reincubation revealed that (-)-isoproterenol inhibited the phosphorylation of a specific protein band with apparent molecular weight of 47,000 (designated band F). These results support the hypothesis that alterations in membrane protein phosphorylation induced by neurotransmitters play a role in the regulation of adenylate cyclase activity.
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002540:Cerebral Cortex / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D007545:Isoproterenol / Q000494:pharmacology*; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008566:Membranes / Q000201:enzymology*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats
chemical_list: D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D000255:Adenosine Triphosphate; D000262:Adenylyl Cyclases; D008274:Magnesium; D007545:Isoproterenol; D002118:Calcium