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1、VIROLOGY179,121-l28(1990)AnalysisoftheSSpike(Peplomer)GlycoproteinofBovineCoronavirusSynthesizedinInsectCellsDONGWANYOO,*+’MICHAELD.PARKER,tANDLORNEA.BABIUK*,t*DepartmentofVeterinaryMicrobiologyandtVeterinaryInfectiousDiseaseOrganization,UniversityofSaskatchewan,124VeterinaryRoa
2、d,Saskatoon,SaskatchewanS7NOWO,CanadaReceivedMarch13.7990;acceptedJune5,1990Thebovinecoronavirus(BCV)spikeglycoproteinprecursor(S,formerlytermedpeplomer)anditstwosubunitpoly-peptides(SlandS2)wereindividuallyexpressedinSpodoprerafrugiperda(Sf9)insectcells.Eachrecombinantbaculovi-
3、rusexpressedbothglycosylated(S,170K;Sl,95K;S2,80K)andunglycosylated(S,,140K;Sl,,75K;andS20,85K)formsofBCVspikepolypeptidesinSf9cells.Themature95KSlpolypeptidewassecretedwhereastheSandS2polypeptidesremainedcell-associated.TheSprecursorwaspartiallycleavedin39cells,andtheresultingS
4、lwasalsoreleasedintothemedium.Neutralizingmonoclonalantibodiesrepresentingtwoantigenicdomainsboundtorecombi-nantSandSlbutnottheS2polvpeotides._.indicatingthattwomajorepitopesforBCVneutralizationarelocatedontheSlsubunit.o1woAcademicPW.S,inc.INTRODUCTIONfusionactivity(Storzetal.,1
5、981;Sturmanetal.,1985).Similartoothercoronaviruses,cleavageoftheBCVSBovinecoronavirus(BCV)isanenteropathogenicglycoproteinispredictedtooccuratthesequenceRR-coronaviruscausingseverediarrheainnewbornSRRinpositions764-768(Parkereta/.,1990a).calves.BCVcontainsfourmajorstructuralprot
6、eins:theTheSglycoproteinfunctionsinvirusattachmenttonucleocapsidprotein(N;50K-54K),thematrixproteinpermissivecells(Collinsetal.,1982;Cavanaghand(M;23K-26K),thehemagglutinin/esterase(HE;124K),Davis,1986),cellfusion(Storzeta/.,1981;Sturmanetandthespikeprotein(S;180K)(KingandBrian,
7、1982;al.,1985;Yooetal.,manuscriptinpreparation),andDeregteta/.,1987;Cry-Coatseta/.,1988).TheHEgly-inductionofneutralizingantibodies(Cavanaghetal.,coproteinisadisulfide-linkeddimer,whichcanbere-1988;Spaaneta/.,1988).Foravianinfectiousbronchi-ducedtotwoidenticalsubunitsof60Kpolype
8、ptide,tisvirus(IBV),themostrelevantregionforint
