N-Glycans and Glycosylphosphatidylinositol-Anchor Act on Polarized Sorting of Mouse PrPC in Madin-Darby Canine Kidney Cells 英文参考文献.docVIP

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N-Glycans and Glycosylphosphatidylinositol-Anchor Act on Polarized Sorting of Mouse PrPC in Madin-Darby Canine Kidney Cells 英文参考文献.doc

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N-Glycans and Glycosylphosphatidylinositol-Anchor Act on Polarized Sorting of Mouse PrPC in Madin-Darby Canine Kidney Cells 英文参考文献

N-GlycansandGlycosylphosphatidylinositol-AnchorAct onPolarizedSortingofMousePrPC inMadin-Darby CanineKidneyCells BertaPuig1,HermannC.Altmeppen1,DanaThurm1,MarkusGeissen1¤,CatharinaConrad1 ,Thomas Braulke2,MarkusGlatzel1* 1InstituteofNeuropathology,UniversityMedicalCenterHamburg-Eppendorf,Hamburg,Germany,2DepartmentofBiochemistry,Children’sHospitalUniversityMedical CenterHamburg-Eppendorf,Hamburg,Germany Abstract The cellular prion protein (PrPC) plays a fundamental role in prion disease. PrPC is a glycosylphosphatidylinositol (GPI)- anchored protein with two variably occupied N-glycosylation sites. In general, GPI-anchor and N-glycosylation direct proteinstoapicalmembranesinpolarizedcellswhereasthemajorityofmousePrPCisfoundinbasolateralmembranesin polarized Madin-Darby canine kidney (MDCK) cells. In this study we have mutated the first, the second, and both N- glycosylationsitesofPrPCandalsoreplacedtheGPI-anchorofPrPCbytheThy-1GPI-anchorinordertoinvestigatetherole ofthesesignalsinsortingofPrPCinMDCKcells.Cellsurfacebiotinylationexperimentsandconfocalmicroscopyshowed thatlackofoneN-linkedoligosaccharideleadstolossofpolarizedsortingofPrPC.ExchangeofthePrPCGPI-anchorforthe oneofThy-1redirectsPrPCtotheapicalmembrane.Inconclusion,bothN-glycosylationandGPI-anchoractonpolarized sortingofPrPC,withtheGPI-anchorbeingdominantoverN-glycans. Citation:PuigB,AltmeppenHC,ThurmD,GeissenM,ConradC,etal.(2011)N-Glycans andGlycosylphosphatidylinositol-AnchorActonPolarizedSortingof MousePrPCinMadin-DarbyCanineKidneyCells.PLoSONE6(9):e24624.doi:10.1371/journal.pone.0024624 Editor:AndrewFrancisHill,UniversityofMelbourne,Australia ReceivedJune16,2011;AcceptedAugust14,2011;PublishedSeptember8,2011 Copyright:?2011Puigetal.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermitsunrestricted use,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding: This research was supported by grants from the De