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1、ArchVirol(1989)106:239-259VirologyArchives©bySpringer-Verlag1989IntracellularsynthesisandprocessingofthestructuralglycoproteinsofturkeyentericcoronavirusS.Dea1,S.Garzon2,andP.Tijssen11CentredeRechercheenM6decineCompar6eUniversit6duQuebec,InstitutArmand-Frappier
2、,Laval-des-Rapides,Quebecand2Facult6deM~decine,Universit6deMontr6al,Montreal,Qu6bec,CanadaAcceptedMarch27,1989Summary.Pulselabelingofcellswith[35S]methionineor[3H]glucosamineatdifferenttimesafterinfection,followedbySDS-PAGEandWesternimmuno-blottinganalysisusing
3、rabbitanti-TCVhyperimmuneserum,wasusedtoresolveandidentifyTCV-inducedintracellularproteins.Theviralstructuralproteins(gp200,gp140/gp66,gpl00/gpl20,p52,andgp24/p20)weredetectedinra-diolabeledcellextractsby9to12hourspost-infection,aswellastwopossiblenon-structura
4、lproteinswithapparentmol.wts,of36,000and32,000.Thepre-dominant52,000nucleocapsidproteincouldbedetectedincelllysatesassoonas6to8hoursafterinfection;itwasinitiallyresolvedasacomplexof3closelymigratingspecieswithmol.wts,rangingfrom46,000to52,000.Pulse-chaseandimmu
5、noprecipitationexperimentsindicatedthatgp200arosefromaputativeprecursorwithmol.wt,of150,000to170,000,thatunderwentglycosylation.Proteolyticcleavageofgp200,inturn,probablyyieldedthegp100andgp120species.TheuniqueTCVhemagglutininproteinoriginatedfromaprimaryprecur
6、sorwithmol.wt,of60,000,whichunderwentrapiddimerizationbydi-sulfidebondformationandglycosylationtoyieldgp140.ThepeplomericandmatrixproteinswerebothshowntobeN-glycosylated,asindicatedbytheirsensitivitytotunicamycin(TM)andtheirresistancetosodiummonensin(SM).Inthep
7、resenceofTM,proteinswithmol.wts,of90,000,120-130,000,and150,000accumulatedinTCV-infectedcellsratherthanpeplomericglycopro-teins,andthematrixproteinE1wasonlydetectedinitsunglycosylatedform.TheadditionofTMtotheculturemediuminterferedwiththematurationofprogenyvira
8、lparticles,assuggestedbytheabsenceofpeplomersatthesurfaceoftheintravacuolarandextracellularvirions,andtheaccumulationofamor-phousmaterialnotfoundintheabsenceoftheglycosylati
