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1、VirusReseurch,4(1986)145-156145ElsevierVRR00232CoronavirusIBVg~ycopolypeptides:locationalstudiesusingproteasesandsaponin,amembranepermeabilizerDavidCavanagh,PhilipJ.Davis1andDarrylJ.C.Pappin’HoughtonPvubryResearchStation,Houghton,Huntingdon,Cum&PEI72DA,and’DepurtmentofBiochemistry,Unic~ers
2、ityofLeeds,LeedsLSZ9JT,U.K.(Acceptedforpublication8October1985)Summary[3sS]methionine-labelledavianinfectiousbronchitisvirus(IBV)(strain41)anditspurifiedproteincomponentsandvirionsofIBV-Beaudettewereincubatedwith10proteases.Severalproteaseshydrolysedallorsomeofthemembraneglycopoly-peptide(
3、M;M,30K)andremovedabout1.3Kofpeptidefromtheamino-(N-)-terminusplusbothglycans,asdeterminedbySDS-polyacrylamidegelelectrophore-sis.N-terminalanalysisof[3H]isoleucine-labeiledMafterhydrolysisbybromelainrevealedthatthefirstnineresidueshadbeenremoved.Afterthevirionshadbeenpermeabilisedwithsapo
4、nin,afurther2.5Kdecreaseinmolecularweightwasproducedandthiswasshowntobefromthecarboxy-(C-)terminus.Whenconsid-eredwiththehydropathicityplotanalysisoftheaminoacidsequenceofM(Boursnell,M.E.G.etal.,1984,VirusRes.1,303-313)theseresultssuggestthatasfewas9-20N-terminalaminoacidresiduesmayprotrud
5、eattheoutermembranesurfaceandthatthereisahighlyproteasesensitivesequenceofanestimated20-25residuesattheC-terminusofMexposedinthelumenofthevirion.S2butnotSlwascleavedtoamajorglycopolypeptideofapproximately71Kbyseveralproteases,andto76Kbytrypsin.N-terminalsequencingofthe71Kglycopolypeptidere
6、vealedthatithadthesameN-terminusasintact52.Afterhydrolysisinthepresenceandabsenceofsaponinitwasconcludedthat52isverysensitivetohydrolysisnearitscarboxyterminusatresiduesclosetotheoutermembranesurface.coronavirusIBV,glycopolypeptides,proteases,saponin0168-1702/g6/$03.50(01986ElsevierScience
7、PublishersB.V.(BiomedicalDivision)146introductionAvianinfectiousbronchitiscoronavirus(IBV)hasthreeproteinstructuralele-ments;aninternalnucleocapsid(N)proteinandtwoenvelope-associatedproteins,themembrane(M)andspike(S)glycoproteins.Preliminarystudiesusingtheprot
