Brain casein kinase 2: affinity purification procedure using immobilized polyethylenimine.
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.
Khilko
S
S
Department of Psychiatry and Behavioral Neurobiology, University of Alabama, Birmingham 35294.
Greenwood
J A
JA
Johnson
G V
GV
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
D047428:Protein Kinase Inhibitors; D011094:Polyethyleneimine; D006493:Heparin; D011494:Protein Kinases; D047388:Casein Kinases