Control of glucokinase translocation in rat hepatocytes by sorbitol and the cytosolic redox state.
In rat hepatocytes cultured in 5 mM glucose, glucokinase activity is present predominantly in a bound state, and during permeabilization of the cells with digitonin in the presence of Mg2+ less than 20% of glucokinase activity is released. However, incubation of hepatocytes with a higher [glucose] [concn. giving half-maximal activation (A50) 15 mM] or with fructose (A50 50 microM) causes translocation of glucokinase from its Mg(2+)-dependent binding site to an alternative site [Agius and Peak (1993) Biochem. J. 296, 785-796]. A comparison of various substrates showed that sorbitol (A50 8 microM) was 6-fold more potent than fructose at causing glucokinase translocation, whereas tagatose was as potent and mannitol was > 10-fold less potent (A50 550 microM). These substrates also stimulate glucose conversion into glycogen with a similar relative potency, suggesting that conversion of glucose into glycogen is dependent on the binding and/or location of glucokinase within the hepatocyte. Ethanol and glycerol inhibited the effects of fructose, sorbitol and glucose on glucokinase translocation, whereas dihydroxy-acetone had a small additive effect at sub-maximal substrate stimulation. The converse effects of glycerol and dihydroxy-acetone suggest a role for the cytosolic NADH/NAD+ redox state in controlling glucokinase translocation. Titrations with three competitive inhibitors of glucokinase did not provide evidence for involvement of glucokinase flux in glucose-induced glucokinase translocation: N-acetylglucosamine inhibited glucose conversion into glycogen, but not glucose-induced glucokinase translocation; glucosamine partially suppressed glucose-induced and fructose-induced glucokinase translocation, at concentrations that caused total inhibition of glucose conversion into glycogen; D-mannoheptulose increased glucokinase release and had an additive effect with glucose. 3,3'-Tetramethylene-glutaric acid (5 mM), an inhibitor of aldose reductase, inhibited glucokinase translocation induced by glucose, but not that by sorbitol or fructose, suggesting that glucose may induce glucokinase translocation by conversion into sorbitol. Sorbitol generated from glucose intrahepatically or extrahepatically in hyperglycaemic conditions may be a physiological regulator of hepatic glucokinase translocation.
Agius
L
L
Department of Medicine, University of Newcastle upon Tyne, U.K.
D000818:Animals; D001692:Biological Transport; D003600:Cytosol / Q000378:metabolism; D004098:Dihydroxyacetone / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology; D005632:Fructose / Q000494:pharmacology; D005941:Glucokinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005977:Glutarates / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D006601:Hexoses / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008112:Liver Glycogen / Q000096:biosynthesis; D008297:Male; D008353:Mannitol / Q000494:pharmacology; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D013012:Sorbitol / Q000378:metabolism*
D005977:Glutarates; D006601:Hexoses; D008112:Liver Glycogen; C060318:tetramethylene glutaric acid; D005632:Fructose; D000431:Ethanol; D008353:Mannitol; D013012:Sorbitol; D005941:Glucokinase; D004098:Dihydroxyacetone; D005990:Glycerol; D010100:Oxygen; C030192:tagatose