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

Y. C. Awasthi, H. S. Garg, D. D. Dao, C. A. Partridge, Satish Srivastava

Research output: Contribution to journalArticle

133 Citations (Scopus)

Abstract

Erythrocyte glutathione (GSH) can be rapidly depleted by incubating the 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 thus 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 hemoglobin 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 also indicate that GSH is vital in maintaining a reduced environment within the erythrocytes.

Original languageEnglish (US)
Pages (from-to)733-738
Number of pages6
JournalBlood
Volume58
Issue number4
StatePublished - 1981
Externally publishedYes

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Dinitrochlorobenzene
Glutathione
Hemoglobins
Erythrocytes
Methemoglobin
Glutathione Transferase
Glutathione Reductase
Acetylcysteine
Glutathione Peroxidase
Oxidation
2,4-dinitrobenzene

ASJC Scopus subject areas

  • Hematology

Cite this

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. / Awasthi, Y. C.; Garg, H. S.; Dao, D. D.; Partridge, C. A.; Srivastava, Satish.

In: Blood, Vol. 58, No. 4, 1981, p. 733-738.

Research output: Contribution to journalArticle

Awasthi, Y. C. ; Garg, H. S. ; Dao, D. D. ; Partridge, C. A. ; Srivastava, Satish. / 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. In: Blood. 1981 ; Vol. 58, No. 4. pp. 733-738.
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AU - Partridge, C. A.

AU - Srivastava, Satish

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