Kinetic properties of Pelophylax esculentus muscle FBPase.
D-Fructose-1,6-bisphosphate 1-phosphohydrolase FBPase; [EC 3.1.3.11] was isolated from Pelophylax esculentus muscle in an electrophoretically homogeneous form with ca 30% yield. Its subunit molecular mass is ca 37 kDa. In this study, we determined the basic kinetic properties of the frog muscle enzyme. FBPase exhibited a maximum activity at pH 7.5. Like other FBPases the frog enzyme requires magnesium ions for its activity (K(a)=263 microM) and is activated by potassium ions (K(a)=63.6 microM). I(0.5) for calcium ion (91 microM) is 100 times higher than the corresponding value of mammalian muscle FBPase. K(s) for the substrate was 1.68 microM. Substrate excess inhibited the enzyme (K(si)=55 microM). AMP and fructose-2,6-bisphosphate (Fru-2,6P(2)) are potent inhibitors of frog muscle FBPase with I(0.5) of 0.2 microM and K(i) of 114 nM, respectively. Both inhibitors act synergistically on the frog muscle FBPase. In the presence of 0.05-0.5 microM of AMP, K(i) for Fru-2,6P(2) is 92 and 28 nM. I(0.5) for AMP for P. esculentus muscle FBPase is 55 times lower than the corresponding value for P. esculentus liver isozyme.
Dziewulska-Szwajkowska
Daria
D
Zoological Institute, University of Wroclaw, ul. Sienkiewicza 21, 50-335 Wroclaw, Poland.
Dzugaj
Andrzej
A
D000249:Adenosine Monophosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006597:Fructose-Bisphosphatase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D005635:Fructosediphosphates / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology*; D011188:Potassium / Q000494:pharmacology; D011898:Ranidae
D004791:Enzyme Inhibitors; D005635:Fructosediphosphates; D000249:Adenosine Monophosphate; C027652:fructose 2,6-diphosphate; D006597:Fructose-Bisphosphatase; D008274:Magnesium; D011188:Potassium; D002118:Calcium