Glutathione oxidation by hypochlorous acid in endothelial cells produces glutathione sulfonamide as a major product but not glutathione disulfide.
Treatment of cells with hypochlorous acid (HOCl) at sublethal doses causes a concentration-dependent loss in reduced glutathione (GSH) levels. We have investigated the products of the reaction of HOCl with GSH in human umbilical vein endothelial cells. Despite a complete loss of GSH, there were only very small increases in intracellular and extracellular glutathione disulfide and glutathione sulfonic acid after exposure to HOCl. (35)S labeling of the GSH pool showed only a minimal increase in protein-bound GSH, suggesting that S-thiolation was not a major contributor to HOCl-mediated loss of GSH in endothelial cells. Rather, the products of the reaction were mostly exported from cells and included a peak that co-eluted with the cyclic sulfonamide that is a product of the reaction of GSH with reagent HOCl. Evidence of this species in endothelial cell supernatants after HOCl treatment was also obtained using electrospray mass spectrometry. In conclusion, exposure to HOCl causes the irreversible loss of cellular GSH with the formation of novel products that are rapidly exported from the cell, and resynthesis of GSH will be required to restore levels. The loss of GSH would alter the redox state of the cell and compromise its defenses against further oxidative stress.
Free Radical Research Group, Department of Pathology, Christchurch School of Medicine, Christchurch, New Zealand. juliet.pullar@chmeds.ac.nz
J M
JM
Pullar
M C
MC
Vissers
C C
CC
Winterbourn
D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000096:biosynthesis*; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology*; D010084:Oxidation-Reduction; D021241:Spectrometry, Mass, Electrospray Ionization; D013450:Sulfones
D013450:Sulfones; C430906:glutathione sulfonamide; D006997:Hypochlorous Acid; D005978:Glutathione; D019803:Glutathione Disulfide