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 proteins to apical membranes in polarized cells whereas the majority of mouse PrPC is found in basolateral membranes in polarized Madin-Darby canine kidney (MDCK) cells. In this study we have mutated the first, the second, and both N-glycosylation sites of PrPC and also replaced the GPI-anchor of PrPC by the Thy-1 GPI-anchor in order to investigate the role of these signals in sorting of PrPC in MDCK cells. Cell surface biotinylation experiments and confocal microscopy showed that lack of one N-linked oligosaccharide leads to loss of polarized sorting of PrPC. Exchange of the PrPC GPI-anchor for the one of Thy-1 redirects PrPC to the apical membrane. In conclusion, both N-glycosylation and GPI-anchor act on polarized sorting of PrPC, with the GPI-anchor being dominant over N-glycans

A Uelhoff

Andrew Francis Hill

B Brugger

B Caughey

B Chesebro

B De Keukeleire

Berta Puig

BR Stevenson

C Korth

C Monnet

Catharina Conrad

CE Le Pichon

D Delacour

D Gravotta

D Sarnataro

DA Brown

Dana Thurm

E Cancellotti

E Grasbon-Frodl

E Stimson

F Goubaeva

FE Cohen

GP Xue

HC Altmeppen

Hermann C. Altmeppen

HR Büeler

J Bremer

J Lauren

J Zuegg

JH Benting

JH Lipschutz

M Geissen

M Glatzel

M Rogers

M Russelakis-Carneiro

Markus Geissen

Markus Glatzel

MG Paulick

MK Salamat

MP Lisanti

N Madore

NL Tuzi

NM Hooper

O Vagin

P Breuer

P Scheiffele

PM Rudd

R Linden

R Mishra

RJ Kascsak

RM Nisbet

S Ikeda

S Lehmann

S Paladino

S Pang

S Paquet

SI Zaidi

SJ DeArmond

T Pan

Thomas Braulke

TW Rademacher

V Beringue

English

PubMed

Crossref

PLoS ONE

N-Glycans and Glycosylphosphatidylinositol-Anchor Act on Polarized Sorting of Mouse PrPC in Madin-Darby Canine Kidney Cells

Puig Berta

Altmeppen Hermann C.

Thurm Dana

Geissen Markus

Conrad Catharina

Braulke Thomas

Glatzel Markus

Public Library of Science (PLOS)

 in Madin-Darby Canine Kidney Cells

The cellular prion protein (PrP(C)) plays a fundamental role in prion disease. PrP(C) is a glycosylphosphatidylinositol (GPI)-anchored protein with two variably occupied N-glycosylation sites. In general, GPI-anchor and N-glycosylation direct proteins to apical membranes in polarized cells whereas the majority of mouse PrP(C) is found in basolateral membranes in polarized Madin-Darby canine kidney (MDCK) cells. In this study we have mutated the first, the second, and both N-glycosylation sites of PrP(C) and also replaced the GPI-anchor of PrP(C) by the Thy-1 GPI-anchor in order to investigate the role of these signals in sorting of PrP(C) in MDCK cells. Cell surface biotinylation experiments and confocal microscopy showed that lack of one N-linked oligosaccharide leads to loss of polarized sorting of PrP(C). Exchange of the PrP(C) GPI-anchor for the one of Thy-1 redirects PrP(C) to the apical membrane. In conclusion, both N-glycosylation and GPI-anchor act on polarized sorting of PrP(C), with the GPI-anchor being dominant over N-glycans

Hermann C Altmeppen

Directory of Open Access Journals

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http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3169634

N-Glycans and Glycosylphosphatidylinositol-Anchor Act on Polarized Sorting of Mouse PrPC in Madin-Darby Canine Kidney Cells

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