Use of immuno-blot techniques to discriminate between the glutathione S-transferase Yf, Yk, Ya, Yn/Yb and Yc subunits and to study their distribution in extrahepatic tissues. Evidence for three immunochemically distinct groups of transferase in the rat

J D Hayes, T J Mantle

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    Abstract

    The glutathione S-transferases are dimeric enzymes whose subunits can be defined by their mobility during sodium dodecyl sulphate/polyacrylamide-gel electrophoresis as Yf (Mr 24,500), Yk (Mr 25,000), Ya (Mr 25,500), Yn (Mr 26,500), Yb1 (Mr 27,000), Yb2 (Mr 27,000) and Yc (Mr 28,500) [Hayes (1986) Biochem. J. 233, 789-798]. Antisera were raised against each of these subunits and their specificities assessed by immuno-blotting. The transferases in extrahepatic tissues were purified by using, sequentially, S-hexylglutathione and glutathione affinity chromatography. Immune-blotting was employed to identify individual transferase polypeptides in the enzyme pools from various organs. The immuno-blots showed marked tissue-specific expression of transferase subunits. In contrast with other subunits, the Yk subunit showed poor affinity for S-hexylglutathione-Sepharose 6B in all tissues examined, and subsequent use of glutathione and glutathione affinity chromatography. Immuno-blotting was employed to identify a new cytosolic polypeptide, or polypeptides, immunochemically related to the Yk subunit but with an electrophoretic mobility similar to that of the Yc subunit; high concentrations of the new polypeptide(s) are present in colon, an organ that lacks Yc.

    Original languageEnglish
    Pages (from-to)779-88
    Number of pages10
    JournalBiochemical Journal
    Volume233
    Issue number3
    DOIs
    Publication statusPublished - 1 Feb 1986

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    Keywords

    • Animals
    • Chromatography, Affinity
    • Cross Reactions
    • Cytosol/metabolism
    • Electrophoresis, Polyacrylamide Gel
    • Glutathione Transferase/analysis
    • Immunoglobulin G/immunology
    • Isoenzymes/analysis
    • Male
    • Peptide Fragments/immunology
    • Proteins/metabolism
    • Rats
    • Rats, Inbred Strains
    • Tissue Distribution

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