Anion-exchange high-performance liquid chromatography of glutathione S-transferases. Separation of the minor isoenzymes of human erythrocyte, heart and lung.

Apparently homogeneous preparation of the acidic glutathione S-transferases were subjected to high-performance liquid chromatography over a SynChropak AX-300 anion-exchange column. Results of high-performance liquid chromatography analysis indicated that in each of these tissues the acidic glutathione S-transferases have one major and two minor isoenzymes. All the isoenzymes obtained by high-performance liquid chromatography are dimers of subunits of molecular weight (Mr) 22,500. The major isoenzymes of all the tissues, representing more than 80% of total glutathione S-transferase activity, have similar retention times and comparable kinetic properties. High-performance liquid chromatography profiles of the lung and heart isoenzymes are similar to each other but are significantly different from that of the erythrocytes. These studies provide evidence of microheterogeneity in the acidic isoenzymes of heart, lung and erythrocyte glutathione S-transferases and provide a rapid and efficient method for separation of minor isoenzymes in more than 95% yield.