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dc.contributor.author이서구-
dc.contributor.author강상원-
dc.date.accessioned2018-05-18T08:15:13Z-
dc.date.available2018-05-18T08:15:13Z-
dc.date.issued2005-
dc.identifier.issn0003-2697-
dc.identifier.otherOAK-2636-
dc.identifier.urihttp://dspace.ewha.ac.kr/handle/2015.oak/243183-
dc.description.abstract2-Cys peroxiredoxin (Prx) is a novel cellular peroxidase that reduces peroxides in the presence of thioredoxin, thioredoxin reductase, and nicotinamide adenine dinucleotide phosphate (NADPH) and that functions in H 2O2-mediated signal transduction. Recent studies have shown that 2-cys Prx can be inactivated by cysteine overoxidation in conditions of oxidative stress. Therefore, peroxidase activity, rather than the protein level, of 2-cys Prx is the more important measure to predict its cellular function. Here, we introduce a modified activity assay method for mammalian 2-cys Prx based on yeast nonselenium thioredoxin reductase. Yeast thioredoxin reductase is expressed in Escherichia coli cells and purified at high yield (40 mg/L of culture broth) as an active flavoprotein by combined diethyl aminoethyl (DEAE) and phenyl hydrophobic chromatography. The optimal concentrations of yeast thioredoxin and thioredoxin reductase required to achieve maximum mammalian 2-cys Prx activity are 3.0 and 1.5 μM, respectively. This modified assay method is useful for measuring 2-cys Prx activity in cell lysates and can also be adapted for a 96-well plate reader for high-throughput screening of chemical compounds that target 2-cys Prx. © 2004 Elsevier Inc. All rights reserved.-
dc.languageEnglish-
dc.titleActivity assay of mammalian 2-cys peroxiredoxins using yeast thioredoxin reductase system-
dc.typeArticle-
dc.relation.issue2-
dc.relation.volume338-
dc.relation.indexSCI-
dc.relation.indexSCIE-
dc.relation.indexSCOPUS-
dc.relation.startpage216-
dc.relation.lastpage236-
dc.relation.journaltitleAnalytical Biochemistry-
dc.identifier.doi10.1016/j.ab.2004.12.008-
dc.identifier.wosidWOS:000227750900006-
dc.identifier.scopusid2-s2.0-14644394199-
dc.author.googleKim J.-A.-
dc.author.googlePark S.-
dc.author.googleKim K.-
dc.author.googleRhee S.G.-
dc.author.googleKang S.W.-
dc.contributor.scopusid이서구(7401852092;25923074700)-
dc.contributor.scopusid강상원(55731433900)-
dc.date.modifydate20190901081003-
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