Brain casein kinase 2: affinity purification procedure using immobilized polyethylenimine.

S Khilko; J A Greenwood; G V Johnson

doi:10.1016/s1046-5928(05)80035-x

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Brain casein kinase 2: affinity purification procedure using immobilized polyethylenimine.

S Khilko·J A Greenwood·G V Johnson

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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.

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Animals Brain Casein Kinases Cattle Chromatography, Affinity Heparin Phosphorylation Polyethyleneimine Protein Kinase Inhibitors Protein Kinases Substrate Specificity Surface Properties Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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S Khilko, J A Greenwood, G V Johnson. 1992. Brain casein kinase 2: affinity purification procedure using immobilized polyethylenimine.. https://doi.org/10.1016/s1046-5928(05)80035-x

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