ENZYMATIC CONJUGATION OF ERYTHROCYTE GLUTATHIONE WITH 1-CHLORO-2,4-DINITROBENZENE - THE FATE OF GLUTATHIONE CONJUGATE IN ERYTHROCYTES AND THE EFFECT OF GLUTATHIONE DEPLETION ON HEMOGLOBIN

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  • 1 January 1981
    • journal article
    • research article
    • Vol. 58  (4) , 733-738
Abstract
Erythrocyte glutathione (GSH) can be rapidly depleted by incubating the human cells with 1-chloro-2,4-dinitrobenzene (CDNB), which forms 2,4-dinitrophenyl-S-glutathione with GSH through the reaction catalyzed by glutathione S-transferase. GSH-CDNB conjugate formed stays undegraded within the erythrocytes. This indicates that in the erythrocytes, mercapturic acid pathway is inoperative. Depletion of GSH in the intact erythrocytes by CDNB results in rapid oxidation of large amounts of Hb to methemoglobin. When glutathione S-transferase-free hemolysate of erythrocytes is incubated with CDNB, the depletion of GSH as well as methemoglobin formation are minimal. Glutathione peroxidase and glutathione reductase activities of the erythrocytes are not affected by CDNB. These studies provide a specific enzymatic method for rapid removal of erythrocyte GSH and indicate that GSH is vital in maintaining a reduced environment within the erythrocytes.