Purification of skeletal muscle hexokinase by affinity elution chromatography
Qadri, S.S.; Easterby, J.S.
Analytical Biochemistry 105(2): 299-303
ISSN/ISBN: 0003-2697 PMID: 7457836 DOI: 10.1016/0003-2697(80)90461-3
Type Ii hexokinase (Ec 188.8.131.52) has been purified from rat skeletal muscle by a simple procedure involving chromatography on DEAE-cellulose, affinity elution chromatography from phosphocellulose, and gel filtration on Sephadex G-200. The key to the preparation of homogeneous enzyme is the affinity elution step in which an effector molecule, glucose 6-phosphate, is used as the eluting ligand. A 5300-fold purification is obtained by the procedure and over 400-fold purification is obtained in the affinity elution step alone. Approximately 3.3 mg of homogeneous hexokinase with a specific activity of 120 units/mg is obtained from 800 g of rat limb.