Analysis of oxidative stress-induced protein carbonylation using fluorescent hydrazides

dc.contributor.authorTamarit Sumalla, Jordi
dc.contributor.authorHoogh, Anouk de
dc.contributor.authorObis Monné, Èlia
dc.contributor.authorAlsina Obiols, David
dc.contributor.authorCabiscol Català, Elisa
dc.contributor.authorRos Salvador, Joaquim
dc.date.accessioned2016-05-18T09:59:14Z
dc.date.embargoEndDate2025-01-01
dc.date.issued2012
dc.description.abstractProtein carbonyl detection has been commonly used to analyze the degree of damage to proteins under oxidative stress conditions. Most laboratories rely on derivatization of carbonyl groups with dinitrophenylhydrazine followed by Western blot analysis using antibodies against the dinitrophenyl moiety. This paper describes a protein carbonyl detection method based on fluorescent Bodipy, Cy3 and Cy5 hydrazides. Using this approach, Western blot and immunodetection are no longer needed, shortening the procedure and increasing accuracy. Combination of Cy3 and Cy5 hydrazides allows multiplexing analyses in a single two-dimensional gel. Derivatization with Bodipy hydrazide allows easy matching of the spots of interest and those obtained by general fluorescent protein staining methods, which facilitates excising target proteins from the gels and identifying them. This method is effective for detecting protein carbonylation in samples of proteins submitted to metal-catalyzed oxidation “in vitro” and assessing the effect of hydrogen peroxide and chronological aging on protein oxidative damage in yeast cells.ca_ES
dc.description.sponsorshipThis work was supported by Grants BFU2010-19193 and CSD2007-00020 from the Ministerio de Ciencia e Innovacion, and SGR0677 from the Generalitat de Catalunya and Fundacio La Marato de TV3 (Spain).ca_ES
dc.identifier.doihttps://doi.org/10.1016/j.jprot.2012.04.046
dc.identifier.idgrec018162
dc.identifier.issn1874-3919
dc.identifier.urihttp://hdl.handle.net/10459.1/57045
dc.language.isoengca_ES
dc.publisherElsevierca_ES
dc.relationMICINN/PN2008-2011/BFU2010-19193
dc.relationMIECI/PN2004-2007/CSD2007-00020
dc.relation.isformatofReproducció del document publicat a https://doi.org/10.1016/j.jprot.2012.04.046ca_ES
dc.relation.ispartofJournal of Proteomics, 2012, vol. 75, núm. 12, p. 3778-3788ca_ES
dc.rights(c) Elsevier, 2012ca_ES
dc.rights.accessRightsinfo:eu-repo/semantics/restrictedAccessca_ES
dc.subjectProtein carbonylsca_ES
dc.subjectOxidative stressca_ES
dc.subjectYeastca_ES
dc.titleAnalysis of oxidative stress-induced protein carbonylation using fluorescent hydrazidesca_ES
dc.typearticleca_ES
dc.type.versionpublishedVersionca_ES
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