Regulatory properties of Rana esculenta liver D-fructose-1,6-bisphosphate 1-phosphohydrolase and their comparison with properties of other vertebrate liver isoenzymes.

D-Fructose-1,6-bisphosphate 1-phosphohydrolase [EC 3.1.3.11] (Fru-1,6P2ase), a regulatory enzyme of gluconeogenesis, was isolated from Rana esculenta liver in homogeneous from with approximately 30% yield. Basic kinetic properties of the enzyme and its subunit molecular weight were determined. Km is 1.72 microM. Like other vertebrate Fru-1,6P2ase, the frog liver enzyme is inhibited by fructose-2,6-bisphosphate (Fru-2,6P2) competitively, Ki is 78 nM and by AMP allosterically, I0.5 is 10.9 microM. Both inhibitors (Fru-2,6P2 and AMP) act synergistically on liver Fru-1,6-P2ase. Ki for Fru-2,6P2 determined in the presence of 1-10 microM of AMP were 35-2 nM, respectively. Maximum activity was found at pH 7.5. Like other Fru-1,6P2ases, the frog enzyme requires magnesium ions for its activity and is activated by potassium ions; the Ka for Mg2+ is 267 microM, Ka for K+ is 77 mM. The subunit molecular weight of the frog liver Fru-1,6P2ase was 37,300 Da. A great similarity between regulatory properties of frog liver Fru-1,6P2ase and liver enzymes of other vertebrates, suggests a similar regulation of gluconeogenesis in amphibia and other vertebrates.