Characterization of alkaline phosphatase from bovine polymorphonuclear neutrophils.
We characterized the bovine polymorphonuclear neutrophil alkaline phosphatase which was considerably purified with a sp. act. of 206 units/mg of protein. The Km value for p-nitrophenylphosphate at pH 10.0 was 1.69 mM. L-Histidine, imidazole and L-homoarginine but not L-phenylalanine inhibited the enzyme. In heat stability study, the enzyme lost 50% activity at 56 degrees C for 20 min. The enzyme had a half-life of 30 min in 3 M urea at 37 degrees C and pH 7.5. The enzyme was inhibited by beta-mercaptoethanol in a dose-dependent fashion. It is suggested from above results that the neutrophil alkaline phosphatase isozyme could be distinguishable from other tissue isozymes.
D000469:Alkaline Phosphatase / Q000097:blood*; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000494:pharmacology; D007700:Kinetics; D007978:Levamisole / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D009504:Neutrophils / Q000201:enzymology*; D013816:Thermodynamics; D014508:Urea / Q000494:pharmacology
D000596:Amino Acids; D007093:Imidazoles; D007978:Levamisole; D008623:Mercaptoethanol; C029899:imidazole; D014508:Urea; D000469:Alkaline Phosphatase