title: Inhibition by the protein kinase C activator 4 beta-phorbol 12-myristate 13-acetate of the prostaglandin F2 alpha-mediated and noradrenaline-mediated but not glucagon-mediated activation of glycogenolysis in rat liver.
abstract: In perfused rat livers, infusion of prostaglandin F2 alpha (PGF2 alpha) or noradrenaline increased glucose and lactate output and reduced flow. Glucagon increased glucose output and decreased lactate output without influence on flow. Infusion of phorbol 13-myristate 14-acetate (PMA) for 20 min prior to these stimuli strongly inhibited the metabolic and hemodynamic effects of noradrenaline, reduced the metabolic actions of PGF2 alpha but did not alter the effects of glucagon. In isolated rat hepatocytes PGF2 alpha, noradrenaline and glucagon activated glycogen phosphorylase but only PGF2 alpha and noradrenaline increased intracellular inositol 1,4,5-trisphosphate (InsP3). The noradrenaline- or PGF2 alpha-elicited activation of glycogen phosphorylase and increase in InsP3 were largely reduced after preincubation of the cells for 10 min with PMA, whereas the glucagon-mediated enzyme activation was not affected. In contrast to PMA, the phorbol ester 4 alpha-phorbol 13,14-didecanoate, which does not activate protein kinase C, did not attenuate the PGF2 alpha- and noradrenaline-elicited stimulation of glucose output, glycogen phosphorylase and InsP3 formation. Stimulation of InsP3 formation by AlF4-, which activates phospholipase C independently of the receptor, was not attenuated by prior incubation with PMA. Plasma membranes purified from isolated hepatocytes had both a high-capacity, low-affinity and a low-capacity, high-affinity binding site for PGF2 alpha. The Kd of the high-capacity, low-affinity binding site was close to the concentration of PGF2 alpha that increased glycogen phosphorylase activity half-maximally. Binding to the high-capacity, low-affinity binding site was enhanced by guanosine 5'-O-(3-thio)triphosphate (GTP[S]). This high-capacity, low-affinity site might thus represent the receptor. The Bmax and Kd of the high-capacity site, as well as the enhancement by GTP[S] of PGF2 alpha binding to this site, remained unaffected by PMA treatment. It is concluded that, in hepatocytes, activation of protein kinase C by PMA interrupted the InsP3-mediated signal pathway from PGF2 alpha via a PGF2 alpha receptor and phospholipase C to glycogen phosphorylase at a point distal of the receptor prior to phospholipase C.
authors.affiliation: Institut für und Molekulare Zellbiologie, Georg-August-Universität, Göttingen, Germany.
authors.forename: G P
authors.initials: GP
authors.lastname: Püschel
authors.forename: H
authors.initials: H
authors.lastname: Miura
authors.forename: F
authors.initials: F
authors.lastname: Neuschäfer-Rube
authors.forename: K
authors.initials: K
authors.lastname: Jungermann
mesh_terms: D017607:Aluminum Compounds / Q000494:pharmacology; D000818:Animals; D015237:Dinoprost / Q000378:metabolism / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D005459:Fluorides / Q000494:pharmacology; D005934:Glucagon / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009638:Norepinephrine / Q000494:pharmacology*; D006005:Phosphorylases / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000378:metabolism
chemical_list: D017607:Aluminum Compounds; D007773:Lactates; D019344:Lactic Acid; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D015544:Inositol 1,4,5-Trisphosphate; D006003:Glycogen; D005934:Glucagon; D015237:Dinoprost; D006005:Phosphorylases; D011493:Protein Kinase C; D010738:Type C Phospholipases; D005947:Glucose; D013755:Tetradecanoylphorbol Acetate; D005459:Fluorides; D009638:Norepinephrine; C032311:aluminum fluoride