Interference by ATPase in the assay of rat heart phosphofructokinase.
A high molecular-weight protein was found in heart extracts which, in the assay for phosphofructokinase, artificially activated the enzyme. The protein could be removed by gel-exclusion chromatography or high-speed centrifugation. The mechanism of activation appeared to be due to the hydrolysis of ATP to ADP, AMP and inorganic phosphate which was inhibited by Mn2+. Phosphofructokinase is thus activated by the production of activators and by the lowered inhibitory concentration of ATP. Since the adrenergic/Ca2+-activated form of the enzyme is the more sensitive to activators, the difference between the two forms of phosphofructokinase is amplified in the presence of the ATPase and diminished upon its removal or its inhibition by Mn2+. The Mn2+-sensitive ATPase appears to play no part in the adrenergic/Ca2+-mediated control of cardiac phosphofructokinase or the interconverting reactions.
D000251:Adenosine Triphosphatases / Q000032:analysis*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002850:Chromatography, Gel / Q000379:methods; D004789:Enzyme Activation / Q000187:drug effects; D004837:Epinephrine / Q000494:pharmacology; D005635:Fructosediphosphates / Q000302:isolation & purification* / Q000502:physiology; D006598:Hexosediphosphates / Q000302:isolation & purification*; D066298:In Vitro Techniques; D008345:Manganese / Q000378:metabolism; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D010732:Phosphofructokinase-1 / Q000032:analysis*; D051381:Rats
D005635:Fructosediphosphates; D006598:Hexosediphosphates; D008345:Manganese; C027652:fructose 2,6-diphosphate; D010732:Phosphofructokinase-1; D000251:Adenosine Triphosphatases; D002118:Calcium; D004837:Epinephrine