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dc.contributor.authorLiang, Xinwen
dc.contributor.authorKaya, Alaattin
dc.contributor.authorZhang, Yan
dc.contributor.authorLe, Dung T
dc.contributor.authorHua, Deame
dc.contributor.authorGladyshev, Vadim N
dc.date.accessioned2013-04-25T14:06:45Z
dc.date.issued2012
dc.identifier.citationLiang, Xinwen, Alaattin Kaya, Yan Zhang, Dung T. Le, Deame Hua, and Vadim N. Gladyshev. 2012. Characterization of methionine oxidation and methionine sulfoxide reduction using methionine-rich cysteine-free proteins. BMC Biochemistry 13:21.en_US
dc.identifier.issn1471-2091en_US
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:10583858
dc.description.abstractBackground: Methionine (Met) residues in proteins can be readily oxidized by reactive oxygen species to Met sulfoxide (MetO). MetO is a promising physiological marker of oxidative stress and its inefficient repair by MetO reductases (Msrs) has been linked to neurodegeneration and aging. Conventional methods of assaying MetO formation and reduction rely on chromatographic or mass spectrometry procedures, but the use of Met-rich proteins (MRPs) may offer a more streamlined alternative. Results: We carried out a computational search of completely sequenced genomes for MRPs deficient in cysteine (Cys) residues and identified several proteins containing 20% or more Met residues. We used these MRPs to examine Met oxidation and MetO reduction by in-gel shift assays and immunoblot assays with antibodies generated against various oxidized MRPs. The oxidation of Cys-free MRPs by hydrogen peroxide could be conveniently monitored by SDS-PAGE and was specific for Met, as evidenced by quantitative reduction of these proteins with Msrs in DTT- and thioredoxin-dependent assays. We found that hypochlorite was especially efficient in oxidizing MRPs. Finally, we further developed a procedure wherein antibodies made against oxidized MRPs were isolated on affinity resins containing same or other oxidized or reduced MRPs. This procedure yielded reagents specific for MetO in these proteins, but proved to be ineffective in developing antibodies with broad MetO specificity. Conclusion: Our data show that MRPs provide a convenient tool for characterization of Met oxidation, MetO reduction and Msr activities, and could be used for various aspects of redox biology involving reversible Met oxidation.en_US
dc.language.isoen_USen_US
dc.publisherBioMed Centralen_US
dc.relation.isversionofdoi:10.1186/1471-2091-13-21en_US
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC3514235/pdf/en_US
dash.licenseLAA
dc.subjectMethionineen_US
dc.subjectMethionine sulfoxideen_US
dc.subjectCysteineen_US
dc.subjectMethionine sulfoxide reductaseen_US
dc.subjectProtein oxidation and reductionen_US
dc.subjectProtein repairen_US
dc.subjectAntibodiesen_US
dc.titleCharacterization of methionine oxidation and methionine sulfoxide reduction using methionine-rich cysteine-free proteinsen_US
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden_US
dc.relation.journalBMC Biochemistryen_US
dash.depositing.authorGladyshev, Vadim N
dc.date.available2013-04-25T14:06:45Z
dc.identifier.doi10.1186/1471-2091-13-21*
dash.contributor.affiliatedKaya, Alaattin
dash.contributor.affiliatedZhang, Yan
dash.contributor.affiliatedGladyshev, Vadim


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