Oxidized glutathione decreases luminal Ca2+ content of the endothelial cell ins(1,4,5)P3-sensitive Ca2+ store.
The model oxidant, t-butyl hydroperoxide (t-buOOH), inhibits Ins(1,4,5)P3-dependent Ca2+ signalling in calf pulmonary artery endothelial cells. Metabolism of t-buOOH within the cytosol is coupled to the oxidation of glutathione. In this study, we investigated whether oxidized glutathione (GSSG) is the intracellular moiety responsible for mediating the effects of t-buOOH on Ca2+ signalling. The increase in cytosolic [Ca2+] stimulated by application of 2,5-di-t-butylhydroquinone (BHQ) was used to estimate the luminal Ca2+ content of the Ins(1,4,5)P3-sensitive store in intact cells. Luminal Ca2+ content was unaffected by t-buOOH (0.4 mM, 0-3 h) unless intracellular GSSG content was concomitantly elevated. The effect was specific for increased GSSG and was not replicated by depletion of GSH. These results suggest that cytosolic GSSG, produced endogenously within the endothelial cell, decreases the luminal Ca2+ content of Ins(1,4,5)P3-sensitive Ca2+ stores. Depletion of internal Ca2+ stores by GSSG may represent a key mechanism by which some forms of oxidant stress inhibit signal transduction in vascular tissue. At the plasma membrane, t-buOOH is known to inhibit the capacitative Ca2+ influx pathway. Increased intracellular GSSG potentiated the inhibitory effect of t-buOOH on Ca2+ influx, thereby providing the first evidence that activity of the capacitative Ca2+ influx channel is sensitive to thiol reagents formed endogenously within the cell.
Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030, USA.
P N
PN
Henschke
S J
SJ
Elliott
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology*; D019328:Buthionine Sulfoximine; D002118:Calcium / Q000378:metabolism*; D002330:Carmustine / Q000494:pharmacology; D002417:Cattle; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D005978:Glutathione / Q000031:analogs & derivatives* / Q000378:metabolism; D019803:Glutathione Disulfide; D006873:Hydroquinones / Q000494:pharmacology*; D015544:Inositol 1,4,5-Trisphosphate / Q000502:physiology*; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology; D010545:Peroxides / Q000378:metabolism / Q000494:pharmacology; D011651:Pulmonary Artery; D015398:Signal Transduction; D020122:tert-Butylhydroperoxide
D000963:Antimetabolites; D000975:Antioxidants; D006873:Hydroquinones; D016877:Oxidants; D010545:Peroxides; D008717:Methionine Sulfoximine; C019359:2,5-di-tert-butylhydroquinone; D019328:Buthionine Sulfoximine; D015544:Inositol 1,4,5-Trisphosphate; D020122:tert-Butylhydroperoxide; D005978:Glutathione; D002118:Calcium; D002330:Carmustine; D019803:Glutathione Disulfide