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1、VirusResearch.5(1986)131-144131ElsevierVRR00262Structuralproteinsofhumanrespiratorycoronavirus0C43BrendaG.HogueandDavidA.BrianDepartmentofMicrobiology,TheUniversityofTennessee,Knoxville,TN379960845,lJ.S.A(Acceptedforpublication28February1986)SummaryThehumanrespiratorycoronavirusOC43wasgrown
2、onahumanrectaltumorcelllineandwasisotopicallylabeledwithaminoacids,glucosamine,andorthophos-phatetoanalyzevirionstructuralproteins.Fourmajorproteinspecieswereresolvedbyelectrophoresisandmanyoftheirpropertieswerededucedfromdigestionstudiesusingproteolyticenzymes.Thefourproteinsare:(1)A190kDa
3、protein,thepresumedpeplomericprotein,thatwasglycosylatedandproteolyticallycleavablebytrypsinintosubunitsof110and90kDa.Thesubunitseachrepresentadifferentaminoacidsequenceonthebasisofpeptidemapping;(2)a130kDaproteinthatwasglycosylatedandbehavedasadisulfide-linkeddimerof65kDamolecules.Itisthea
4、pparentvirionhemagglutininonthebasisofdigestionstudieswithtrypsin,bromelainandpronase;(3)a55kDanucleocapsidproteinthatwasphosphorylated;(4)a26kDamatrixproteinthatwasglycosylated.The190,130,55and26kDaspeciescanthereforebedesignatedP,H,NandM,respectively.Theyexistinmolarratiosof4:1:33:33,anda
5、recalculatedtobepresentattherateof88,22,726,and726moleculespervirion,respectively.humanrespiratorycoronovirus0C43,structuralproteinsIntroductionCoronavirusesareestimatedtocauseapproximately20%ofupperrespiratorydiseaseinhumans(Larsenetal.,1980;McIntoshetal.,1970).Humanrespiratorycoronaviruse
6、sarefoundaseitheroneoftwoantigenictypes:thoserelatedtohumancoronavirus0C43(virusesthatcouldoriginallybegrownonlyintrachealorganculture),andthoserelatedtohumancoronavirus229E(virusesthatcouldbe0168-1702/86/$03.5001986ElsevierSciencePublishersB.V.(BiomedicalDivision)132isolatedintissueculture
7、)(McIntoshetal.,1970).Arigorouscharacterizationofthestructuralproteinsofhumanrespiratorycoronavirusesmaythereforerequireaseparatefocusoneachantigenicsubgroup.Allcoronavirusesstudiedindetailtodateappeartoshareacommongenomestructure,namely,asingle-stranded