Formation and cleavage of mixed disulfide hemoglobin glutathione in intact erythrocytes

A marked decrease in the level of GSH was observed in human erythrocytes incubated with glucose in the presence of acetyl phenylhydrazine. All the GSH lost could not be accounted for as GSSG. After washing and reincubation of the erythrocytes with phosphate saline containing glucose, almost 80% of the GSH could be recovered. The rate of appearance of GSH was much faster when inosine and adenine were also incorporated in the incubation medium. The results of these studies, taken together with previous studies of GSH Hb mixed disulfide, suggest that in the presence of acetyl phenylhydrazine, a portion of GSH oxidized becomes bound to haemoglobin or other proteins in the form of mixed disulfide. GSH from the mixed disulfide is released in the presence of NADPH and glutathione reductase.