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1、VIROLOGY77,650-660(1977)CharacterizationofaCoronavirus’II.GlycoproteinsoftheViralEnvelope:TrypticPeptideAnalysisLAWRENCES.STURMAN*ANDKATHRYNV.HOLMES3DivisionofLaboratoriesandResearch,NewYorkStateDepartmentofHealth,Albany,NewYork12201andDepartmentofMic
2、robiology,UniversityofTexas,SouthwesternMedicalSchool,Dallas,Texas75235AcceptedNovember29,1976Twospeciesofmembrane-associatedglycoproteinshavebeenidentifiedinthecorona-virusvirion.Theyarereadilydistinguishedonthebasisofsize,radiolabelingcharacter-isti
3、cs,andlocationinrelationtothelipidbilayer.Thelargerglycoproteinishighlylabeledbybothradiolabeledfucoseandglucosamine.Thisspeciesisfoundintwoforms,GP180andGP90,withapparentmolecularweightsof180,000and90,000.GP180canbeconvertedtoGP90invitrobytreatmentof
4、virionswithtrypsin.AnalysisoftrypticdigestsofGP90andGPl80giveidenticalpeptidepatterns.Basedonpronaseandbromelainsensitiyities,GP180/90istheonlyproteinwhichislocatedentirelyexternaltotheviralenvelope.Itappearstocomprisethecharacteristiclong,petal-shape
5、dpeplom-ersofthevirion.Thesmallerglycoprotein,GP23,hasanapparentmolecularweightof23,000andislabeledbyradiolabeledglucosaminebutnotbyfucose.Thelevelofglucosamine-labelingofGP23isaboutl/10thatofGP180/90.GP23appearstopossesstwodistinctdomains:asmaller,ca
6、rbohydratecontainingregionwhichisfoundoutsidetheviralenvelope,andalargerportion,highlylabeledbymethionine,whichisinte-grallyassociatedwiththeviralmembrane.Anewnomenclatureisproposedforthethreemajorcoronavirusstructuralproteins.Thetwoenvelopeglycoprote
7、ins,GP23andGP180/90aredesignatedElandE2,respectively;theinnercoreprotein,~~50,isdesignatedN.INTRODUCTIONThestructuralproteinsofthecoronavi-Coronaviruseshaveadistinctiveap-rusA59virionhavebeenidentifiedbyso-pearance(McIntosh,1974).Innegativelydiumdodec
8、ylsulfate-polyacrylamidegelstainedpreparationstheexternalsurfaceelectrophoresis(SDS-PAGE)(&u-man,ofthevirionenvelopeiscoveredwithpe-1977).Understandardconditions,sixma-plomers,20nmlong,whicharewidelyjorpolypeptidespeciesarefoundwithap-spacedan
