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1、VIROLOGY77,637-649(19’i’i’)CharacterizationofaCoronavirus’I.StructuralProteins:EffectsofPreparativeConditionsontheMigrationofProteininPolyacrylamideGelsLAWRENCES.STURMAN2DivisionofLaboratoriesandResearch,NewYorkStateDepartmentofHealth,Albany,NewYork12201AcceptedNovember29,1976CoronavirusA59pos
2、sessesfoursizeclassesofstructuralproteinswhichhaveappar-entmolecularweightsmeasuredbySDS-polyacrylamidegelelectrophoresis(SDS-PAGE)of23,000(GP23),50,000(VP50),90,000(GP90),and180,000(GP1801.VP50istheonlystructuralproteinwhichiscompletelyunaffectedbyproteasetreatmentofintactvirions.Thisspeciesi
3、sthemosthighlylabeledbypolaraminoacidssuchasglutamicacidandarginineanditisprobablyassociatedwiththeviralnucleocapsid.GP90,GP180,andGP23aremembrane-associatedproteins.However,afterproteasetreatmentofvirions,only20%oftheGP23moleculeisdigested,whereasalloftheGP90andGP180areremoved.GP90andGPl80app
4、eartocomprisemostoftheprominentlayerofcharacteristicprojectionsontheexternalsurfaceoftheviralenvelope.ThemajorportionofGP23ispresumedtoliewithinthelipidenvelope,protectedfromproteasedigestion.GP23andtheproteaseresistantportion,p*18,exhibitanomalousbehavioronSDS-PAGE.Afterheatingto100”inSDSthee
5、lectrophoreticmobilityofthesepolypep-tidesisalteredandseveralnewformsoflowermobilityareproduced.P-Mercaptoetha-no1anddithiothreitolexaggeratetheeffectsofheating.INTRODUCTIONwhiletheothersareassociatedwiththeCoronavirusesaremorphologicallydis-viralenvelope.Althoughsixspeciesaretinctiveenveloped
6、RNAviruseswhichbudconsistentlyresolvedundertheusualcon-fromcytoplasmicmembranessuchasen-ditionsofgelelectrophoresis,ourstudiesdoplasmicreticulum,cytoplasmicvesiclessuggestthatseveralofthesespeciesareandtheGolgiapparatus(McIntosh,1974).differentformsofthesamemolecule.Whilecoronaviruseshavebeens
7、tudiedex-Thereappeartobefoursizeclassesoftensivelymorphologically,theyhavenotstructuralproteinswhichhavebeendesig-beenwell-characterizedchemically.UsingnatedGP180,GP90,VP50,andGP23.themurinecoronavirusA59,ananalysisGP23willbeshowntoexhi
