Alkaline phosphatase from pig kidney. Method of purification and molecular properties.
Alkaline phosphatase (EC 3.1.3.1) from pig kidney brush-border membranes was solubilized from membrane precipitates by butan-1-ol at a critical pH of 7.0. The 12000-fold purification procedure included (NH(4))(2)SO(4) precipitation, DEAE-and TEAE-cellulose chromatography, Sephadex G-200 gel filtration and neuraminidase digestion followed by DEAE-cellulose chromatography. The purified protein contained 20% (w/w) carbohydrate and had mol.wt. 150000-156000 as estimated by Sephadex filtration and ultracentrifuge analysis. It was a tetrameric glycoprotein consisting of identical subunits, and it had a molecular activity at 25 degrees C of 2600s(-1) per tetramer. Its concentration in kidney was estimated to be 8.5-8.8mg/kg.
D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000818:Animals; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007122:Immunoelectrophoresis; D007668:Kidney / Q000201:enzymology*; D008566:Membranes / Q000201:enzymology; D008970:Molecular Weight; D013053:Spectrophotometry; D013552:Swine; D013696:Temperature
D006023:Glycoproteins; D000469:Alkaline Phosphatase