Rat red blood cell hexokinase purification, properties and age-dependence.
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.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004905:Erythrocyte Aging; D004912:Erythrocytes / Q000201:enzymology*; D006593:Hexokinase / Q000097:blood* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007527:Isoenzymes; D008970:Molecular Weight; D051381:Rats; D012156:Reticulocytes / Q000201:enzymology*
D007527:Isoenzymes; D006593:Hexokinase