Purification of a novel 30,000 Da calcium-binding protein from bovine cerebellum.
A novel very acidic calcium-binding protein (CaBP) was purified from bovine cerebellum, using 45Ca autoradiography as a marker, through a preparative procedure involving salting out with a very high concentration of ammonium sulfate, DE52 column chromatography, RNAase treatment, and HPLC gel filtration. This protein showed a molecular weight of 30,0000 dalton (Da) on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and of 120,000 on in gel filtration chromatography analysis under physiological ionic strength. The calcium binding activity of this 30,000 Da CaBP was monitored on the basis of calcium-dependent changes in tyrosine fluorescence (Kd = 3.0 microM).
Kurokawa
H
H
Department of Pharmacology, Faculty of Medicine, University of Tokyo.
Nonomura
Y
Y
D000818:Animals; D002135:Calcium-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002531:Cerebellum / Q000378:metabolism*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D007700:Kinetics; D008970:Molecular Weight
D002135:Calcium-Binding Proteins