Inhibition of hexokinase activity by a fructose 2,6-bisphosphate-dependent cytosolic protein from liver.
Mammalian and yeast hexokinases were found to be reversibly inhibited by fructose 2,6-bisphosphate, an effect requiring the presence of a cytosolic protein factor. Experimental evidence suggests that this factor (inhibitor) is a regulatory protein, the interactions of which with hexokinases are modulated by fructose 2,6-bisphosphate. The Vmax of hexokinase D was decreased, and no changes on other kinetic parameters were observed. The inhibitor was present in fresh liver cytosol filtered through Sephadex G-25 and was partially isolated by negative absorption on DEAE-cellulose followed by ammonium sulfate fractionation. The inhibitor was also present in brain and kidney, but not in muscle. A molecular mass of 200,000 was determined by gel filtration. The inhibition was dependent on the concentrations of both the inhibitory protein and fructose 2,6-bisphosphate. No delay in fructose 2,6-bisphosphate inhibition was observed. Several other hexose phosphates were tested and were not effective. In the presence of amounts of inhibitor sufficient to produce complete inhibition of hexokinase D, the concentration of fructose 2,6-bisphosphate required to produce 50% inhibition was about 0.5 microM. The inhibitor was unstable and was stabilized by the presence of fructose 2,6-bisphosphate.
D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000302:isolation & purification; D005635:Fructosediphosphates / Q000494:pharmacology*; D006593:Hexokinase / Q000037:antagonists & inhibitors*; D006598:Hexosediphosphates / Q000494:pharmacology*; D006600:Hexosephosphates / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D011506:Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D051381:Rats; D013696:Temperature
D004791:Enzyme Inhibitors; D005635:Fructosediphosphates; D006598:Hexosediphosphates; D006600:Hexosephosphates; D007527:Isoenzymes; D011506:Proteins; C027652:fructose 2,6-diphosphate; D006593:Hexokinase