The separation of glutathione transferase subunits by using reverse-phase high-pressure liquid chromatography

Abstract

A simple method is described for the separation and quantification of the subunits of GSH transferases present in rat tissue extracts. This method, involving GSH-agarose affinity chromatography followed by reverse-phase h.p.l.c., is rapid and sufficiently sensitive to measure 5 micrograms of each subunit in a mixture. Examples are given of its application to extracts of rat kidney, adrenal, testicular interstitial cells and seminiferous tubules. The analysis of seminiferous tubules indicates that the technique may be of value for the identification of novel subunits. Preliminary separations of subunits from human GSH transferases are also described.

Polymorphism of the glutathione transferase subunit 3 in Sprague–Dawley rats involves a reactive cysteine residue

“…Individual peaks were identified as GST subunits as shown in Figure 1 by SDS\PAGE, and immunoblotting with anti-GST antibodies (results not shown). The elution pattern of SD rat liver GSTs ( Figure 1A) was consistent with previous reports [34,35]. However, in the case of HHR liver GSTs ( Figure 1B), instead of subunit 3, a peak named X was detected between GST subunits 3 and 4.…”

“…Individual GST subunits were separated by HPLC, essentially as described by Ostlund-Farrants et al [34]. HPLC was carried out using a TSK-Gel Octadecyl-4PW column (4.6 mmi150 mm ; Tosoh, Tokyo, Japan).…”

Glutathione S‐Transferase, Similar to Sigma Class, from Skin Secretions of Xenopus laevis

“…Reversed-phase HPLC analysis of af nitypuri ed GST was performed by the method of Ostlund Farrants et al (23) with an HPLC-Kontron system equipped with a rheodyne injector and a photodiode variable-wavelength detector. Samples of GST were injected onto a reverse phase column Vydak 201 TP 5 (200 £ 3 mm) (Chrompack, Middelburg, The Netherlands) that had been equilibrated with 35% acetonitrile in 0.1% (v/v) tri uoroacetic acid.…”

Cloning, sequencing and characterization of the human Alpha glutathione S-transferase gene corresponding to the cDNA clone pGTH2

“…DISCUSSION H.p.l.c. data revealed that two GST Alpha isoenzymes are present in human liver and kidney tissue [5,7]. Recently, we showed that the kidney isoenzymes, al and a2, have retention times similar to the two liver forms, referred to as ax and ay [5].…”

“…analysis demonstrates two GST Alpha subunits in liver and kidney, referred to as a. and ay or a, and a2 respectively; the former is more basic than the latter [5,7], the pl being 8.9 for al and 8.4 for a2 [1].…”