title: Brain casein kinase 2: affinity purification procedure using immobilized polyethylenimine.
abstract: A simplified procedure for casein kinase 2 purification from bovine brain is described. The purification procedure consists of two affinity chromatography steps, using heparin and polyethylenimine immobilized on a synthetic matrix (Toyopearl 650M). The adsorption and elution conditions for each column were optimized, resulting in a simple elution protocol for each column. A stable, highly purified casein kinase 2 preparation was obtained in 4 h using this procedure. Polyethylenimine was shown to stimulate the casein kinase 2 activity using exogeneous substrates (casein, calmodulin, MAP2, and tau) but not the enzyme's autophosphorylation activity. The polyethylenimine stimulation could be overcome by applying a mass excess of the casein kinase 2 inhibitor, heparin.
authors.affiliation: Department of Psychiatry and Behavioral Neurobiology, University of Alabama, Birmingham 35294.
authors.forename: S
authors.initials: S
authors.lastname: Khilko
authors.forename: J A
authors.initials: JA
authors.lastname: Greenwood
authors.forename: G V
authors.initials: GV
authors.lastname: Johnson
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology*; D047388:Casein Kinases; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D006493:Heparin; D010766:Phosphorylation; D011094:Polyethyleneimine; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000302:isolation & purification*; D013379:Substrate Specificity; D013499:Surface Properties
chemical_list: D047428:Protein Kinase Inhibitors; D011094:Polyethyleneimine; D006493:Heparin; D011494:Protein Kinases; D047388:Casein Kinases