Purification of 3-phosphoglycerate kinase from diverse sources by affinity elution chromatography

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Abstract
Affinity elution chromatography was used to purify phosphoglycerate kinase [EC 2.7.2.3] from different sources. The choice of buffer pH for the chromatography was made according to the relative electrophoretic mobility of the enzyme from the species concerned. Outlines of the methods used to isolate the enzyme from over 20 sources are presented. The enzyme was purified from the muscle tissue of many mammals, fish and birds, from liver of several animals, from yeast, Escherichia coli and plant leaves. The more acidic varieties of the enzymes were purified by conventional gradient elution from ion-exchanges as affinity elution procedures were not applicable. The structural and kinetic parameters investigated show that phosphoglycerate kinase is evolutionarily a highly conservative enzyme; there were few differences in properties regardless of source or function (glycolytic, gluconeogenic or photosynthetic). A detailed comparison of the enzyme preparations purified from bovine muscle and bovine liver failed to detect any significant differences between them. The evidence indicates that they are genetically identical.