Purification of vitamin D-dependent 28,000-Mr calcium-binding protein from bovine cerebellum and kidney by calcium-dependent elution from DEAE-cellulose DE-52 column chromatography.
A vitamin D-dependent calcium-binding protein of 28,000 Mr was purified by a convenient method from the high-ammonium sulfate-saturated fraction (70 to 100% saturation) of bovine cerebellum and kidney. This method is based on the calcium-dependent DEAE-cellulose column chromatography that reflected the specific feature of this protein: different eluting salt concentrations in the presence or absence of Ca2+. The procedure of purification was easily performed by the detection of calcium-binding protein using 45Ca autoradiography. The purified calcium-binding protein showed the same molecular weight and calcium-dependent change of mobility in nondenaturing gel electrophoresis as vitamin D-dependent calcium-binding protein. The 28,000-Mr calcium-binding protein was induced in kidney by the injection of vitamin D in vitamin D-deficient rats.
D000818:Animals; D002135:Calcium-Binding Proteins / Q000302:isolation & purification*; D002417:Cattle; D002531:Cerebellum / Q000032:analysis*; D002848:Chromatography, DEAE-Cellulose; D007668:Kidney / Q000032:analysis*; D008970:Molecular Weight; D009928:Organ Specificity; D064030:S100 Calcium Binding Protein G / Q000302:isolation & purification*; D014808:Vitamin D Deficiency / Q000378:metabolism
D002135:Calcium-Binding Proteins; D064030:S100 Calcium Binding Protein G