Role of amino acid oxidation and protein unfolding in peroxyl radical and peroxynitrite-induced inactivation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides

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dc.catalogadorgjm
dc.contributor.authorFigueroa Alegría, Juan David
dc.contributor.authorFuentes Lemus, Eduardo Felipe
dc.contributor.authorReyes Valenzuela, Juan Sebastián
dc.contributor.authorLoaiza Hernández, Matías Ignacio
dc.contributor.authorAliaga Miranda, Margarita Elly
dc.contributor.authorFierro Huerta, Angélica
dc.contributor.authorLeinisch, Fabian
dc.contributor.authorHagglund, Per
dc.contributor.authorDavies, Michael J.
dc.contributor.authorLópez Alarcón, Camilo Ignacio
dc.date.accessioned2025-01-02T16:18:15Z
dc.date.available2025-01-02T16:18:15Z
dc.date.issued2022
dc.description.abstractThe mechanisms underlying the inactivation of Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase (G6PDH) induced by peroxyl radicals (ROO center dot) and peroxynitrite (ONOO-), were explored. G6PDH was incubated with AAPH (2,2'-azobis(2-methylpropionamidine)dihydrochloride), used as ROO center dot source, and ONOO-. Enzymatic activity was assessed by NADPH generation, while oxidative modifications were analyzed by gel electrophoresis and liquid chromatography (LC) with fluorescence and mass detection. Changes in protein conformation were studied by circular dichroism (CD) and binding of the fluorescent dye ANS (1-anilinonaph-thalene-8-sulfonic acid). Incubation of G6PDH (54.4 mu M) with 60 mM AAPH showed an initial phase without significant changes in enzymatic activity, followed by a secondary time-dependent continuous decrease in activity to similar to 59% of the initial level after 90 min. ONOO- induced a significant and concentration-dependent loss of G6PDH activity with similar to 46% of the initial activity lost on treatment with 1.5 mM ONOO-. CD and ANS fluorescence indicated changes in G6PDH secondary structure with exposure of hydrophobic sites on exposure to ROO center dot, but not ONOO-. LC-MS analysis provided evidence for ONOO--mediated oxidation of Tyr, Met and Trp residues, with damage to critical Met and Tyr residues underlying enzyme inactivation, but without effects on the native (dimeric) state of the protein. In contrast, studies using chloramine T, a specific oxidant of Met, provided evidence that oxidation of specific Met and Trp residues and concomitant protein unfolding, loss of dimer structure and protein aggregation are involved in G6PDH inactivation by ROO center dot. These two oxidant systems therefore have markedly different effects on G6PDH structure and activity.
dc.fuente.origenWOS
dc.identifier.doi10.1016/j.freeradbiomed.2022.08.010
dc.identifier.eissn1873-4596
dc.identifier.issn0891-5849
dc.identifier.urihttp://doi.org/10.1016/j.freeradbiomed.2022.08.010
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/89451
dc.identifier.wosidWOS:000859405900004
dc.information.autorucEscuela de Química; Figueroa Alegría, Juan David; S/I; 1071087
dc.information.autorucEscuela de Química; Fuentes Lemus, Eduardo Felipe; 0000-0002-1465-8466; 186720
dc.information.autorucEscuela de Química; Reyes Valenzuela, Juan Sebastián; S/I; 1268701
dc.information.autorucEscuela de Química; Loaiza Hernández, Matías Ignacio; S/I; 1027529
dc.information.autorucEscuela de Química; Aliaga Miranda, Margarita Elly; 0000-0002-4143-0301; 13361
dc.information.autorucEscuela de Química; Fierro Huerta, Angélica; 0000-0002-6507-4188; 218137
dc.information.autorucEscuela de Química; López Alarcón, Camilo Ignacio; S/I; 1004308
dc.language.isoen
dc.nota.accesocontenido parcial
dc.pagina.final306
dc.pagina.inicio292
dc.revistaFree Radical Biology and Medicine
dc.rightsacceso restringido
dc.subjectGlucose 6-phosphate dehydrogenase
dc.subjectPeroxyl radicals
dc.subjectPeroxynitrite
dc.subjectChloramine T
dc.subjectMethionine oxidation
dc.subjectTyrosine oxidation
dc.subjectTryptophan oxidation
dc.subjectEnzymatic activity
dc.subjectProtein unfolding
dc.titleRole of amino acid oxidation and protein unfolding in peroxyl radical and peroxynitrite-induced inactivation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides
dc.typeartículo
dc.volumen190
sipa.codpersvinculados1071087
sipa.codpersvinculados186720
sipa.codpersvinculados1268701
sipa.codpersvinculados1027529
sipa.codpersvinculados13361
sipa.codpersvinculados218137
sipa.codpersvinculados1004308
sipa.trazabilidadWOS;2022-10-11
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