Bovine kidney alkaline phosphatase. Purification, subunit structure, and metalloenzyme properties.

Bovine Kidney Alkaline Phosphatase

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...

Purification and properties of bovine synovial fluid alkaline phosphatase.

Alkaline phosphatase from bovine synovial fluid was purified 2300-fold. A molecular weight of 72,300 was determined from sucrose density gradient studies. The following monoesters were hydrolyzed by the enzyme: P-glycerophosphate, galactosamine 6-phosphate, glucosamice 6-phosphate, glucose 6-phosphate, o-phospho-L-serine, o-carboxyphenyl phosphate, phenyl phosphate, and p-nitrophenyl phosphate....

Purification of kidney alkaline phosphatase.

Purification of Alkaline Phosphatase

8. An increase in the rate of progression of the bands down the column decreases the sharpness of the bands. On 'Zeo-Karb 215' (40-60 mesh/in.) a rate ofprogression of 10-15 cm./hr. gave satisfactory results. 9. Equations have been derived permitting the calculation ofthe proportion ofthe column occupied by a component, the width ofthe boundaries and the expected yield of pure components in sep...

Purification and properties of alkaline phosphatase from rat chloroma.

Activity was determined by the rate of hydrolysis of 13-glycerophosphate on p-nitrophenyl phosphate (8). The determination with sodium 13-glycerophosphate was carried out as follows. The reaction mixture consisted of 0.05 M Tnis-HC1 (pH 10), 0.023 M sodium 13-glycerophosphate, 0.004 M MgCl2, and enzyme in a final volume of 2 ml. After incubation at 37° for 30 mm, 2 ml of 10% tnicbloroacetic ac...