Brain casein kinase 2: Affinity purification procedure using immobilized polyethylenimine

Sergey Khilko, Jeffrey A. Greenwood, Gail V.W. Johnson

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)355-361
Number of pages7
JournalProtein Expression and Purification
Volume3
Issue number5
DOIs
StatePublished - Oct 1992
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology

Fingerprint

Dive into the research topics of 'Brain casein kinase 2: Affinity purification procedure using immobilized polyethylenimine'. Together they form a unique fingerprint.

Cite this