title: Glutathione and glutaredoxin act as a backup of human thioredoxin reductase 1 to reduce thioredoxin 1 preventing cell death by aurothioglucose.
abstract: Thioredoxin reductase 1 (TrxR1) in cytosol is the only known reductant of oxidized thioredoxin 1 (Trx1) in vivo so far. We and others found that aurothioglucose (ATG), a well known active-site inhibitor of TrxR1, inhibited TrxR1 activity in HeLa cell cytosol but had no effect on the viability of the cells. Using a redox Western blot analysis, no change was observed in redox state of Trx1, which was mainly fully reduced with five sulfhydryl groups. In contrast, auranofin killed cells and oxidized Trx1, also targeting mitochondrial TrxR2 and Trx2. Combining ATG with ebselen gave a strong synergistic effect, leading to Trx1 oxidation, reactive oxygen species accumulation, and cell death. We hypothesized that there should exist a backup system to reduce Trx1 when only TrxR1 activity was lost. Our results showed that physiological concentrations of glutathione, NADPH, and glutathione reductase reduced Trx1 in vitro and that the reaction was strongly stimulated by glutaredoxin1. Simultaneous depletion of TrxR activity by ATG and glutathione by buthionine sulfoximine led to overoxidation of Trx1 and loss of HeLa cell viability. In conclusion, the glutaredoxin system and glutathione have a backup role to keep Trx1 reduced in cells with loss of TrxR1 activity. Monitoring the redox state of Trx1 shows that cell death occurs when Trx1 is oxidized, followed by general protein oxidation catalyzed by the disulfide form of thioredoxin.
authors.affiliation: Division of Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institute, 17177 Stockholm, Sweden.
authors.forename: Yatao
authors.initials: Y
authors.lastname: Du
authors.forename: Huihui
authors.initials: H
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Lu
authors.forename: Arne
authors.initials: A
authors.lastname: Holmgren
mesh_terms: D000818:Animals; D001310:Auranofin / Q000494:pharmacology; D006051:Aurothioglucose / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology; D015153:Blotting, Western; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D054477:Glutaredoxins / Q000378:metabolism*; D005978:Glutathione / Q000037:antagonists & inhibitors / Q000378:metabolism*; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D054833:Isoindoles; D008954:Models, Biological; D009154:Mutation; D016566:Organoselenium Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D054481:Thioredoxin Reductase 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*; D013997:Time Factors
chemical_list: D001393:Azoles; D054477:Glutaredoxins; D054833:Isoindoles; D016566:Organoselenium Compounds; D017382:Reactive Oxygen Species; D006051:Aurothioglucose; D001310:Auranofin; C042986:ebselen; D019328:Buthionine Sulfoximine; D013879:Thioredoxins; D054481:Thioredoxin Reductase 1; D005978:Glutathione