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dc.contributor.authorJobbagy, Soma
dc.contributor.authorVitturi, Dario A
dc.contributor.authorSalvatore, Sonia R
dc.contributor.authorTurell, Lucía
dc.contributor.authorPires, Maria F
dc.contributor.authorKansanen, Emilia
dc.contributor.authorBatthyany, Carlos
dc.contributor.authorLancaster, Jack R Jr
dc.contributor.authorFreeman, Bruce A
dc.contributor.authorSchopfer, Francisco
dc.date.accessioned2019-02-04T14:25:47Z
dc.date.available2019-02-04T14:25:47Z
dc.date.issued2019
dc.identifier.urihttps://erepo.uef.fi/handle/123456789/7430
dc.description.abstractCells evolved robust homeostatic mechanisms to protect against oxidation or alkylation by electrophilic species. Glutathione (GSH) is the most abundant intracellular thiol, protects cellular components from oxidation and is maintained in a reduced state by glutathione reductase (GR). Nitro oleic acid (NO2-OA) is an electrophilic fatty acid formed under digestive and inflammatory conditions that both reacts with GSH and induces its synthesis upon activation of Nrf2 signaling. The effects of NO2-OA on intracellular GSH homeostasis were evaluated. In addition to upregulation of GSH biosynthesis, we observed that NO2-OA increased intracellular GSSG in an oxidative stress-independent manner. NO2-OA directly inhibited GR in vitro by covalent modification of the catalytic Cys61, with kon of (3.45 ± 0.04) × 103 M−1 s−1, koff of (4.4 ± 0.4) × 10−4 s−1, and Keq of (1.3 ± 0.1) × 10−7 M. Akin to NO2-OA, the electrophilic Nrf2 activators bardoxolone-imidazole (CDDO-Im), bardoxolone-methyl (CDDO-Me) and dimethyl fumarate (DMF) also upregulated GSH biosynthesis while promoting GSSG accumulation, but without directly inhibiting GR activity. In vitro assays in which GR was treated with increasing GSH concentrations and GSH depletion experiments in cells revealed that GR activity is finely regulated via product inhibition, an observation further supported by theoretical (kinetic modeling of cellular GSSG:GSH levels) approaches. Together, these results describe two independent mechanisms by which electrophiles modulate the GSH/GSSG couple, and provide a novel conceptual framework to interpret experimentally determined values of GSH and GSSG.
dc.language.isoenglanti
dc.publisherElsevier BV
dc.relation.ispartofseriesRedox biology
dc.relation.urihttp://dx.doi.org/10.1016/j.redox.2018.11.008
dc.rightsCC BY-NC-ND https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectglutathione
dc.subjectglutathione reductase
dc.subjectelectrophile
dc.subjectnitrated fatty acid
dc.subjectdisulfide
dc.subjectNrf2
dc.subjectthiol
dc.subjectoxidation-reduction (redox)
dc.titleElectrophiles modulate glutathione reductase activity via alkylation and upregulation of glutathione biosynthesis
dc.description.versionpublished version
dc.contributor.departmentA.I. Virtanen -instituutti
uef.solecris.id59843470en
dc.type.publicationTieteelliset aikakauslehtiartikkelit
dc.rights.accessrights© Authors
dc.relation.doi10.1016/j.redox.2018.11.008
dc.description.reviewstatuspeerReviewed
dc.publisher.countryAlankomaat
dc.relation.articlenumber101050
dc.relation.volume21
dc.rights.accesslevelopenAccess
dc.type.okmA1
uef.solecris.openaccessOpen access -julkaisukanavassa ilmestynyt julkaisu


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