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Protein Expr Purif. 1992 Oct;3(5):355-61.

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

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  • 1Department of Psychiatry and Behavioral Neurobiology, University of Alabama, Birmingham 35294.

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.

PMID:
1458049
[PubMed - indexed for MEDLINE]

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