N-glycans and glycosylphosphatidylinositol-anchor act on polarized sorting of mouse PrP(C) in Madin-Darby canine kidney cells.
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N-glycans and glycosylphosphatidylinositol-anchor act on polarized sorting of mouse PrP(C) in Madin-Darby canine kidney cells. / Puig Martorell, Berta; Altmeppen, Hermann C.; Thurm, Dana Kathrin; Geissen, Markus; Conrad, Catharina; Braulke, Thomas; Glatzel, Markus.
In: PLOS ONE, Vol. 6, No. 9, 9, 2011, p. 24624.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - N-glycans and glycosylphosphatidylinositol-anchor act on polarized sorting of mouse PrP(C) in Madin-Darby canine kidney cells.
AU - Puig Martorell, Berta
AU - Altmeppen, Hermann C.
AU - Thurm, Dana Kathrin
AU - Geissen, Markus
AU - Conrad, Catharina
AU - Braulke, Thomas
AU - Glatzel, Markus
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
KW - Animals
KW - Mice
KW - Dogs
KW - Protein Transport
KW - Cell Line
KW - Microscopy, Confocal
KW - Biotinylation
KW - Glycosylphosphatidylinositols/metabolism
KW - Polysaccharides/metabolism
KW - PrPC Proteins/metabolism
KW - Animals
KW - Mice
KW - Dogs
KW - Protein Transport
KW - Cell Line
KW - Microscopy, Confocal
KW - Biotinylation
KW - Glycosylphosphatidylinositols/metabolism
KW - Polysaccharides/metabolism
KW - PrPC Proteins/metabolism
U2 - 10.1371/journal.pone.0024624
DO - 10.1371/journal.pone.0024624
M3 - SCORING: Journal article
VL - 6
SP - 24624
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 9
M1 - 9
ER -
