Kinetic properties of pig (Sus scrofa domestica) and bovine (Bos taurus) D-fructose-1,6-bisphosphate 1-phosphohydrolase (F1,6BPase): liver-like isozymes in mammalian lung tissue.
F1,6BPases from porcine and bovine lung were isolated and their kinetic properties were determined. Ks, Kis and beta were determined assuming partial-noncompetitive inhibition (simple intersecting hyperbolic noncompetitive inhibition) of the enzyme by the substrate. Values for Ks were 4.1 and 4.4 microM for porcine and bovine F1,6BPase, respectively and values for 1 were close to 0.55 in both cases. Kis were 9 and 15 microM for porcine and bovine F1,6BPase, respectively. I0.5 for AMP were determined as 7 microM for pig enzyme and 14 microM for F1,6BPase from bovine lung. The enzymes were inhibited by F2,6BP with Ki's of 0.19 and 0.21 microM for porcine and bovine enzymes, respectively. In the presence of AMP concentration equal to I0.5, the Ki values for pig and bovine enzymes were 0.07 and 0.09 microM, respectively. The levels of F2,6BP, AMP and antioxidant enzymes activities in pig and bovine lung tissues were also determined. The cDNA coding sequence of pig lung F1,6BPase1 showed a high homology with pig liver enzyme, differing only in four positions (G/C-63, T/A-808, G/C-884 and T/A-1005) resulting in a single amino acid substitution (Gly-295 for Ala-295). It is hypothesized that the lung F1,6BPase participates in gluconeogenesis, surfactant synthesis and antioxidant reactions.
Rakus
D
D
Department of Animal Physiology, Zoological Institute, Wroclaw University, Cybulskiego, Poland.
Skalecki
K
K
Dzugaj
A
A
D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D002374:Catalase / Q000378:metabolism; D002417:Cattle; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D006597:Fructose-Bisphosphatase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D005952:Glucose-6-Phosphatase / Q000378:metabolism; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism; D005980:Glutathione Reductase / Q000378:metabolism; D020128:Inhibitory Concentration 50; D007527:Isoenzymes; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D008954:Models, Biological; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000378:metabolism; D019764:Phosphoenolpyruvate Carboxykinase (ATP) / Q000378:metabolism; D010734:Phosphogluconate Dehydrogenase / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013379:Substrate Specificity; D013482:Superoxide Dismutase / Q000378:metabolism; D013552:Swine
D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D000249:Adenosine Monophosphate; D010088:Oxidoreductases; D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D002374:Catalase; D013482:Superoxide Dismutase; D005980:Glutathione Reductase; C020667:glutathione oxidase; D006597:Fructose-Bisphosphatase; D005952:Glucose-6-Phosphatase; D019764:Phosphoenolpyruvate Carboxykinase (ATP); D008274:Magnesium