Investigation on the mechanism by which fructose, hexitols and other compounds regulate the translocation of glucokinase in rat hepatocytes.
In isolated hepatocytes in suspension, the effect of sorbitol but not that of fructose to increase the concentration of fructose 1-phosphate and to stimulate glucokinase was abolished by 2-hydroxymethyl-4-(4-N,N-dimethylamino-1-piperazino)-pyrimidine (SDI 158), an inhibitor of sorbitol dehydrogenase. In hepatocytes in primary culture, fructose was metabolized at approximately one-quarter of the rate of sorbitol, and was therefore much less potent than the polyol in increasing the concentration of fructose 1-phosphate and the translocation of glucokinase. In cultures, sorbitol, commercial mannitol, fructose, D-glyceraldehyde or high concentrations of glucose caused fructose 1-phosphate formation and glucokinase translocation in parallel. Commercial mannitol was contaminated by approx. 1% sorbitol, which accounted for its effects. The effects of sorbitol, fructose and elevated concentrations of glucose were partly inhibited by ethanol, glycerol and glucosamine. Mannoheptulose increased translocation without affecting fructose 1-phosphate concentration. Kinetic studies performed with recombinant human beta-cell glucokinase indicated that this sugar, in contrast with N-acetylglucosamine, binds to glucokinase competitively with the regulatory protein. All these observations indicate that translocation is promoted by agents that favour the dissociation of the glucokinase-regulatory-protein complex either by binding to the regulatory protein (fructose I-phosphate) or to glucokinase (glucose, mannoheptulose). They support the hypothesis that the regulatory protein of glucokinase acts as an anchor for this enzyme that slows down its release from digitonin-permeabilized cells.
Niculescu
L
L
Laboratoire de Chimie Physiologique, Université Catholique de Louvain, Brussels, Belgium.
Veiga-da-Cunha
M
M
Van Schaftingen
E
E
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D004098:Dihydroxyacetone / Q000494:pharmacology; D000431:Ethanol / Q000494:pharmacology; D005632:Fructose / Q000494:pharmacology*; D005636:Fructosephosphates / Q000378:metabolism; D005941:Glucokinase / Q000378:metabolism*; D005985:Glyceraldehyde / Q000494:pharmacology; D006601:Hexoses / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D007064:L-Iditol 2-Dehydrogenase / Q000037:antagonists & inhibitors*; D008099:Liver / Q000201:enzymology*; D008297:Male; D008353:Mannitol / Q000378:metabolism; D008356:Mannoheptulose / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar
D005636:Fructosephosphates; D006601:Hexoses; D011743:Pyrimidines; C103900:SDI 158; C032284:fructose-1-phosphate; D005632:Fructose; D005985:Glyceraldehyde; D000431:Ethanol; D008353:Mannitol; D008356:Mannoheptulose; D007064:L-Iditol 2-Dehydrogenase; D005941:Glucokinase; D004098:Dihydroxyacetone; C030192:tagatose; D000117:Acetylglucosamine