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.