冠狀病毒Coronavirusl論文-1991 Residues involved in the antigenic sites of transmissible gastroenteritis coronavirus S glycoprotein.pdf

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1、VIROLOGY183,225-238(1991)ResiduesInvolvedintheAntigenicSitesofTransmissibleGastroenteritisCoronavirusSGlycoproteinFiiTIMAGEBAUER,*WILLEMP.A.POSTHUMUS,tISABELCORREA,*CARLOSSUl%,*CRISTIANSMERDOU,*CARLOSM.SANCHEZ,*JOHANNESA.LENSTRA,+ROBH.MELOEN,tANDLUISENJUANES*,*Centrod

2、eBiologiaMolecular,CSIC-UniversidadAukkoma,CantoBlanco,28049Madrid,Spain;tCenfra/VeterinaryInstitute,P.O.Box65,8200At3Lelystad,TheNetherlands;andHnstituteofInfectiousDiseasesandImmunology,VeterinaryFaculty,StateUniversity,P.O.Box80,165,3508TDUtrecht,TheNetherlandsRece

3、ivedDecember28,1990;acceptedApril1,1991TheSglycoproteinoftransmissiblegastroenteritisvirus(TGEV)hasbeenshowntocontainfourmajorantigenicsites(A,B,C,andD).SiteAisthemaininducerofneutralizingantibodiesandhasbeenpreviouslysubdividedintothethreesubsitesAa,Ab,andAC.Theresid

4、uesthatcontributetothesesiteswerelocalizedbysequenceanalysisof21mutantsthatescapedneutralizationorbindingbyTGEV-specificmonoclonalantibodies(MAbs),andbyepitopescanning(PEPSCAN).SiteAcontainstheresidues538,591,and543,whichareessentialintheformationofsubsitesAa,Ab,andAC

5、,respectively.Inaddition,marmutant1B.H8withresidue586changedhadpartiallyalteredbothsubsiteAaandAb,indicatingthatthesesubsitesoverlapinresidue586;i.e.thisresiduealsoispartofsiteA.Thepeptide537-MKSGYGQPIA-547represents,atleastpartially,subsiteACwhichishighlyconservedamo

6、ngcoronaviruses.Thissiteisrelevantfordiagnosisandcouldbeofinterestforprotection.OtherresiduescontributetositeB(residues97and144),siteC(residues50and51),andsiteD(residue385).ThelocationofsiteDisinagreementwithPEPSCANresults.SiteCcanberepresentedbythepeptide48-P-P/S-N-S

7、-D/E-52butisnotexposedonthesurfaceofnativevirus.ItsaccessibilitycanbemodulatedbytreatmentatpH>l1(at4)andtemperatures>45.SitesAandBarefullydepen-dentonglycosylationforproperfolding,whilesitesCandDarefullyorpartiallyindependentofglycosylation,respec-tively.OncetheSglyco

8、proteinhasbeenassembledintothevirion,thecarbohydratemoietyisnotessentialfortheantigenicsites.01991AcademicPress,hINTRODUCTIO