Abstract
Alkaline phosphatase from swine kidney cortex was purified by a series of procedures; butanol extraction, isoelectric and acetone precipitation, ammonium sulfate fractionation and also ion-exchange, concanavalin A (Con A) affinity chromatography and gel filtration. Elution profile of Con A affinity chromatography showed that the enzyme contains a carbohydrate moiety which reacts with Con A. The purified enzyme was found to be a homogeneous glycoprotein having a molecular weight of 160, 000 by polyacrylamide gel- and immunoelectrophoresis. The result that the molecular weight of the enzyme decreased to one half that of native molecule after heated in sodium dodecyl sulfate (SDS) solution indicates a dimer structure of the enzyme comprising two similar or identical subunits of 80, 000 daltons.
The rabbit antiserum against the swine kidney alkaline phosphatase cross-reacted to phosphatases of bovine enamel organ and human fibrous dysplasia bone.
