Partial purification of alkaline phosphatase from bovine polymorphonuclear neutrophils and some properties.
Alkaline phosphatase was purified from bovine polymorphonuclear neutrophils by butanol extraction and a combination of ion exchange, gel filtration and affinity chromatography. The enzyme was partially purified 2300-fold with a 4.7% yield and a sp. act. of 206 units/mg of protein. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate indicated a single activity band with the mol. wt of 165,000. The pH optima for the enzyme were 10.0 with p-nitrophenylphosphate and phenylphosphate and were 9.0 when beta-glycerophosphate, AMP and ADP were used. The enzyme was activated by Mg2+, Mn2+, Co2+ and Ni2+ but was inhibited by Zn2+. The enzyme was inhibited by EDTA and the EDTA-inactivated enzyme was reactivated by Mg2+, Mn2+ and Co2+ but not Zn2+.
D000469:Alkaline Phosphatase / Q000097:blood* / Q000302:isolation & purification; D000818:Animals; D002413:Cations, Divalent; D002417:Cattle; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009504:Neutrophils / Q000201:enzymology*
D002413:Cations, Divalent; D004492:Edetic Acid; D000469:Alkaline Phosphatase