A NEW HUMAN SELENIUM-CONTAINING PROTEIN.pdf
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A NEW HUMAN SELENIUM-CONTAINING PROTEIN.pdf

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THEJOURNALOFBIOLOGICALCHEMISTRYVol.273,No.15,IssueofApril10,pp.8910–8915,1998PrintedinU.S.A.ANewHumanSelenium-containingProteinPURIFICATION,CHARACTERIZATION,ANDcDNASEQUENCE*(Receivedforpublication,October6,1997,andinrevisedform,February2,1998)VadimN.Gladyshev‡§¶,Kuan-TehJeangi,JohnC.Wootton**,andDolphL.Hatfield‡‡‡Fromthe‡SectionontheMolecularBiologyofSelenium,BasicResearchLaboratory,NCI,the§LaboratoryofBiochemistry,NHLBI,theiLaboratoryofMolecularMicrobiology,NIAID,andthe**NationalCenterforBiotechnologyInformation,NationalLibraryofMedicine,NationalInstitutesofHealth,Bethesda,Maryland20892Seleniumwhichoccursinproteinsastheaminoacid,andbiochemicalstudies(1,2).Thistraceelementisanaturalselenocysteine,isessentialfornumerousbiologicalcomponentofseveralprokaryoticandeukaryoticproteins.Al-processesandforhumanhealth.Aprominent75Se-thoughseleniumoccursinprokaryoticproteinseitherasalabeledproteindetectedinhumanT-cellsmigratedasacofactororasaselenocysteineresidue,mammalianselenopro-15-kDabandbySDS-polyacrylamidegelelectrophore-teinsidentifiedthusfarcontainseleniumonlyintheformofsis.Thisproteinsubunitwaspurifiedandsubjectedtoselenocysteine,whichisthe21stnaturallyoccurringaminotrypticdigestionandpeptidesequenceanalyses.Se-acidinprotein.AselenocysteinetRNAthatdecodesUGAhasquencesoftrypticpeptidesderivedfromtheproteinbeenfoundinalllifekingdoms,suggestingthattheuseofUGAcorrespondedtoahumanplacentalgenesequencecon-asacodonforselenocysteineiswidespreadinnature(2).TheDownloadedfromtaininganopenreadingframeof162residuesandaspecialconservedstem-loopstructuresinthe39-untranslatedreadthroughin-frameTGAcodon.Threedifferentpep-regionsofmammalianselenoproteinmRNAsareessentialfortidesequencesofthe15-kDaproteincorrespondedtoarecognitionofUGAasacodonforselenocysteine,ratherthananucleotidesequencelocateddownstreamofthiscodon,codonforterminationoftranslation(3).suggestingthattheT-cell15-kDaselenoproteincontainsaselenocysteineresidueencodedbyTGA.Post-transla-Oftheeleve