Bovine kidney alkaline phosphatase. Purification, subunit structure, and metalloenzyme properties.
Kidney alkaline phosphatase was purified to homogeneity. It is a glycoprotein of about 172,000 molecular weight. Analyses of the subunit structure by sedimentation equilibrium in 6 M guanidine hydrochloride and by gel electrophoresis in sodium dodecyl sulfate indicate that the alkaline phosphatase is a dimer comprising two very similar or identical subunits of about 87,000 molecular weight. The native enzyme contains 4.5 +/- 0.2 g atoms of zinc per mol of protein. Reconstitution experiments from the apophosphatase show that binding of 4 Zn2+ per mol of dimer is essential for full activity. The kinetic data of Zn2+ binding to the apoprotein require at least a two-step mechanism, in which one of the steps corresponds to a conformational change within the enzyme. This paper also presents data concerning amino acid composition, sugar content, enzyme stability, absorbance index, and sedimentation velocity.
D000469:Alkaline Phosphatase / Q000302:isolation & purification / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001059:Apoproteins / Q000302:isolation & purification / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004589:Electrophoresis, Disc; D006595:Hexosamines / Q000032:analysis; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008667:Metalloproteins / Q000302:isolation & purification / Q000378:metabolism; D008970:Molecular Weight; D009439:Neuraminidase; D012794:Sialic Acids / Q000032:analysis; D015032:Zinc / Q000032:analysis
D000596:Amino Acids; D001059:Apoproteins; D002241:Carbohydrates; D006595:Hexosamines; D008667:Metalloproteins; D012794:Sialic Acids; D000469:Alkaline Phosphatase; D009439:Neuraminidase; D015032:Zinc