title: cAMP-dependent protein kinase activation lowers hepatocyte cAMP.
abstract: Rat hepatocyte protein kinase was activated by incubating the cells with various cAMP analogs. Boiled extracts were then prepared and Sephadex G-25 chromatography was carried out. The G-25 procedure separated the analogs from cAMP since the resin had the unexpected property of binding cyclic nucleotides with differing affinities. Separation was necessary because the analogs would otherwise interfere with the sensitive protein kinase activation method developed for assay of cAMP. The cAMP analogs, but not 5'-AMP, lowered basal cAMP by 50-70%. The effect was rapid, analog concentration-dependent, and occurred parallel with phosphorylase activation, suggesting that the cAMP analogs act through cAMP-dependent protein kinase activation. A cAMP analog completely blocked the cAMP elevation produced by relatively low concentrations of glucagon, but did not block the phosphorylase response, indicating that the cAMP analog substitutes for cAMP as the intracellular activator of protein kinase. One implication of the results is that elevation of cAMP and protein kinase activity by hormones has a negative feedback effect on the cellular cAMP level.
authors.forename: J D
authors.initials: JD
authors.lastname: Corbin
authors.forename: S J
authors.initials: SJ
authors.lastname: Beebe
authors.forename: P F
authors.initials: PF
authors.lastname: Blackmore
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000032:analysis; D000818:Animals; D002850:Chromatography, Gel; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000032:analysis*; D006152:Cyclic GMP / Q000032:analysis; D004789:Enzyme Activation; D005934:Glucagon / Q000494:pharmacology; D008099:Liver / Q000032:analysis*; D008297:Male; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D005934:Glucagon; D000242:Cyclic AMP; D011494:Protein Kinases; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D006152:Cyclic GMP