Separation and characterization of hexokinase I subtypes from human erythrocytes

G Rijksen, G Jansen, R J Kraaijenhagen, M J Van der Vlist, A M Vlug, G E Staal

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23 Citations (Scopus)

Abstract

Hexokinase (ATP: D-hexose 6-phosphotransferase, EC 2.7.1.1) type 1 from human erythrocytes exists in four electrophoretical distinct forms, termed Ia, Ib, Ic and Id in order of their increasing anodal electrophoretic mobility at pH 8.8. We were able to separate type Ia, Ib and Icd on phosphocellulose by using a discontinuous gradient elution. The three chromatographically distinct forms do not differ in their affinity constants for the substrates glucose and MgATP2-. In addition the inhibition by glucose 1,6-diphosphate does not differ significantly for all forms. However, the regulation of these inhibitions by inorganic phosphate is much less for type Ia compared to the other subtypes (P = 0.001). Aging of the red cells is accompanied by a relative increase of the proportion of type Ic and Ia, which is the less regulated form of the enzyme. This shift in electrophoretic and regulatory properties is argued to be due to a post-translational modification of the primary enzyme.

Original languageEnglish
Pages (from-to)292-301
Number of pages10
JournalBiochimica et Biophysica Acta. Reviews on Cancer
Volume659
Issue number2
Publication statusPublished - 15 Jun 1981

Keywords

  • Adenosine Triphosphate/metabolism
  • Cation Exchange Resins
  • Cell Survival
  • Cellulose/analogs & derivatives
  • Chromatography, Ion Exchange
  • Electrophoresis, Cellulose Acetate
  • Erythrocytes/enzymology
  • Glucose-6-Phosphate/analogs & derivatives
  • Glucose/metabolism
  • Glucosephosphates/pharmacology
  • Hexokinase/blood
  • Humans
  • Isoenzymes/blood

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