Rabbit red blood cell hexokinase. Purification and properties.
Rabbit red blood cell hexokinase (EC 2.7.1.1.) has been purified 300,000-fold by a combination of ion exchange chromatography, affinity chromatography, and preparative polyacrylamide gel electrophoresis. The hexokinase activity has been isolated in 35% yield as a protein that is homogeneous by polyacrylamide and sodium dodecyl sulfate gel electrophoresis. The highest specific activity obtained was 145 units/mg of proteins. The native protein has a molecular weight of 110,000 by gel filtration on Ultrogel AcA 44 and 112,000 by sedimentation velocity on sucrose density gradients. Sodium dodecyl sulfate-polyacrylamide gels gave a molecular weight of 110,000 indicating that hexokinase is a monomer. The enzyme had a pI of 6.20 to 6.30 pH units by isoelectric focusing. The enzyme was specific for Mg . ATP and Mg . ITP as the nucleotide substrates. Several hexokinase with different affinities.
D000255:Adenosine Triphosphate; D000818:Animals; D004912:Erythrocytes / Q000201:enzymology*; D006593:Hexokinase / Q000097:blood* / Q000302:isolation & purification; D007293:Inosine Triphosphate; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D011817:Rabbits; D013379:Substrate Specificity
D007293:Inosine Triphosphate; D000255:Adenosine Triphosphate; D006593:Hexokinase; D008274:Magnesium