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1、Articlepubs.acs.org/JACSSemisyntheticLectin−4-DimethylaminopyridineConjugatesforLabelingandProfilingGlycoproteinsonLiveCellSurfaces†††‡††TakahiroHayashi,YediSun,TomonoriTamura,KeikoKuwata,ZhiningSong,YousukeTakaoka,,†andItaruHamachi*†DepartmentofSyntheticChemistryandBiologicalChemis
2、try,GraduateSchoolofEngineering,KyotoUniversity,Katsura,Kyoto615-8510,Japan‡InstituteofTransformativeBio-Molecules(WPI-ITbM),NagoyaUniversity,Chikusa,Nagoya464-8601,Japan*SSupportingInformationABSTRACT:Glycoproteinsoncellsurfacesplayimportantrolesinbiologicalprocesses,includingcell
3、−cellinteraction/signaling,immuneresponse,andcelldifferentiation.Giventhediversityofthestructureofglycans,labelingandimagingofselectedglycoproteinsarechallenging,althoughseveralpromisingstrategieshavebeendevelopedrecently.Here,wedesignandconstructsemisyntheticreactivelectins(sugar-b
4、indingproteins)thatareabletoselectivelylabelglycoproteins.CongerinII,ananimalgalectin,andwheatgermagglutininareconjugatedwith4-dimethylaminopyridine(DMAP),awell-knownacyltransfercatalystbyouraffinity-guidedDMAPmethodandCu(I)-assistedclickchemistry.Selectivelabelingofglycoproteinsisfa
5、cilitatedbytheDMAP-tetheredlectincatalystsbothinvitroandonlivingcells.Two-dimensionalpolyacrylamidegelelectrophoresis(2D-PAGE)analysisenabledustoisolatelabeledglycoproteinsthatareuniquelyexposedondistinctcelllines.Furthermore,thecombinationofimmunoprecipitationwithmassspectrometry(
6、MS)-fingerprintingtechniquesallowedustocharacterize48glycoproteinsendogenouslyexpressedonHeLacells,andsomelow-abundantglycoproteins,suchasepidermalgrowthfactorreceptor(EGFR)andneuropilin-1,weresuccessfullyidentified.OurresultsdemonstratethatsemisyntheticDMAP-tetheredlectinsprovideane
7、wtoolforlabelingandprofilingglycoproteinsonlivingcells.■INTRODUCTIONcontainingreactivehandles(e.g.,azide,alkyne,orketone/aldehydegroups),followedbybioorthogonalprobeligation,Glycoproteinsplayimportantrolesinnumerousbiologicalwhichenabledselectivelabelingofglycansonlivingcellsandproc
8、esses,suchascell−cellinter
