Purification of rat hepatic glucokinase.
We have developed a rapid, reliable procedure for the purification of rat hepatic glucokinase. The purification utilizes DEAE-cellulose, two affinity chromatography steps, and high-performance liquid chromatography. Glucokinase with a specific activity of 240 units/mg, a 42 K-fold purification, and a yield of 60% is obtained. The enzyme appears as a homogeneous band, with over 99% purity as assessed by polyacrylamide gel electrophoresis. The purification procedure can be completed in 5 days.
D000818:Animals; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005941:Glucokinase / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D051381:Rats
D005941:Glucokinase