Selective elution of soluble rat liver glutathione transferases from a glutathione-Sepharose affinity column.
Glutathione transferases (GST) are dimeric enzymes that take part in many detoxification processes. A previous report described the use of a glutathione-Sepharose affinity matrix for the purification of human liver GST. The method involved the use of 5 mM glutathione in a high pH buffer, and the yields were nearly 100%. This method and adapted techniques have now been applied to rat liver GST. Selective GST elution can be obtained in several different ways: by stepwise change of the pH and/or glutathione concentration, and by linear gradient elution. Gel electrophoresis showed, however, that none of the fractions contained pure GST isoenzymes. Also, less than 50% of the total rat liver GST was eluted with 5 mM glutathione, in contrast to the results with human liver GST. A glutathione concentration of 30 mM is necessary for quantitative desorption of rat liver GST from a glutathione-Sepharose column.
Dierickx
P J
PJ
Instituut voor Hygiëne en Epidemiologie, Brussels, Belgium.
D000818:Animals; D002021:Buffers; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005978:Glutathione; D005982:Glutathione Transferase / Q000302:isolation & purification*; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012685:Sepharose
D002021:Buffers; D007527:Isoenzymes; D012685:Sepharose; D005982:Glutathione Transferase; D005978:Glutathione