Open AccessInvolvement of Phosphorylase Kinase Inhibition in the Effect of Resorcinol and Proglycosyn on Glycogen Metabolism in the Liver
Author(s) -
Schaftingen Emile
Publication year - 1995
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1995.301_c.x
Subject(s) - glycogen phosphorylase , chemistry , biochemistry , resorcinol , metabolism , phosphorylase kinase , hepatocyte , phosphatase , glycogen , intracellular , kinase , glycogen synthase , enzyme , organic chemistry , in vitro
The purpose of this study was to identify the mechanism by which proglycosyn and resorcinol decrease the phosphorylase a content and the fructose 2,6‐bisphosphate concentration in isolated hepatocytes. The intracellular concentrations of the glucuronide derivatives of proglycosyn and resorcinol have been measured by HPLC in hepatocytes incubated for 5 min or 30 min with different concentrations of these agents. At both times, there was a reciprocal relationship between the phosphorylase a content and the intracellular concentration of the glucuronidated metabolites, half‐maximal inactivation being observed at about 2 μmol/g protein and 0.25 μmol/g protein for resorcinylglucuronide and proglycosyn‐glucuronide, respectively. Glycogen synthase was not significantly activated by these agents after 5 min but was well activated after 30 min. Preincubation of hepatocytes with 1 mM resorcinol or with 100 μM proglycosyn resulted in a decrease in the rate at which phosphorylase was activated following the addition of glucagon, vasopressin, the protein phosphatase inhibitor calyculin A or the calcium ionophore A 23187, but did not reduce the rate of synthase inactivation. Proglycosynglucuronide and resorcinylglucuronide inhibited phosphorylase kinase in liver Sephadex filtrates, with K i values of about 0.75 mM and 4 mM, respectively. Preincubation of the filtrates with ATP and cAMP decreased the sensitivity of phosphorylase kinase to resorcinylglucuronide by about fourfold. It is concluded that the effect of resorcinol and proglycosyn on the phosphorylase a content is due, at least partly, to an inhibition of phosphorylase kinase by their glucuronidated metabolites. Resorcinol and proglycosyn caused a parallel decrease in the concentration of fructose 2,6–bisphosphate and of hexose 6‐phosphates, without significantly changing the activity of 6‐phosphofructo‐2‐kinase. The decrease in the fructose 2,6‐bisphosphate concentration appears therefore to be secondary to the decrease in the hexose 6‐phosphate concentration.