Rat red blood cell hexokinase purification, properties and age-dependence

✍ Scribed by Giordano Serafini; Mauro Magnani; Vilberto Stocchi; Marina Dachà; Giorgio Forniani

• Publisher: Springer
• Year: 1986
• Tongue: English
• Weight: 467 KB
• Volume: 69
• Category: Article
• ISSN: 0300-8177
• DOI: 10.1007/bf00224765

✦ Synopsis

Rat erythrocytes, in contrast to red blood cells from other mammals, have been shown to contain only one hexokinase isozymic form identified as type I by chromatographic and kinetic properties. Rat reticulocytes contain 3.6-times the hexokinase activity found in mature erythrocytes but exactly the same isozyme. By a combination of ion-exchange chromatography, dye-ligand chromatography and high-pressure liquid chromatography the rat erythrocyte hexokinase was purified more than 84 000-fold to a specific activity of 143 units/mg protein and shown to be homogeneous by sodium dodecyl sulfate-gel electrophoresis. The native protein showed a molecular weight of 100 000 by gel-filtration and an apparent molecular weight of 98 000 under denaturating conditions in sodium dodecyl sulfate-gel electrophoresis. The isoelectric point was shown to be 6.3 pH units. This data provides evidence of only one form of hexokinase in the erythrocytes of a mammal.