Involvement of glypican-1 autoprocessing in scrapie infection.
The copper-binding cellular prion protein (PrP(C)) and the heparan sulphate (HS)-containing proteoglycan glypican-1 (Gpc-1) can both be attached to lipid rafts via their glycosylphosphatidylinositol anchors, and copper ions stimulate their cointernalization from the cell surface to endosomes. The prion protein controls cointernalization and delivers copper necessary for S-nitrosylation of conserved cysteines in the Gpc-1 core protein. Later, during recycling through endosomal compartments, nitric oxide can be released from the S-nitroso groups and catalyses deaminative degradation and release of the HS substituents. Here, by using confocal immunofluorescence microscopy, we show that normal PrP(C) and Gpc-1 colocalize inside GT1-1 cells. However, in scrapie-infected cells (ScGT1-1), Gpc-1 protein remained at the cell surface separate from the cellular prion protein. Scrapie infection stimulated Gpc-1 autoprocessing and the generated HS degradation products colocalized with intracellular aggregates of the disease-related scrapie prion protein isoform (PrP(Sc)). Coimmunoprecipitation experiments demonstrated an association between Gpc-1 and PrP(C) in uninfected cells, and between HS degradation products and PrP(Sc) in infected cells. Silencing of Gpc-1 expression or prevention of Gpc-1 autoprocessing elevated the levels of intracellular PrP(Sc) aggregates in infected cells. These results suggest a role for Gpc-1 autoprocessing in the clearance of PrP(Sc) from infected cells.
Löfgren
Kajsa
K
Department of Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden. lofgren@dbb.su.se
Cheng
Fang
F
Fransson
Lars-Ake
LA
Bedecs
Katarina
K
Mani
Katrin
K
D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000503:physiopathology; D002451:Cell Compartmentation / Q000502:physiology; D002457:Cell Extracts; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004705:Endocytosis / Q000502:physiology; D053673:Glypicans / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000378:metabolism*; D018621:PrPC Proteins / Q000378:metabolism; D018620:PrPSc Proteins / Q000378:metabolism; D011328:Prions / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology; D012608:Scrapie / Q000378:metabolism* / Q000503:physiopathology
D002457:Cell Extracts; D053673:Glypicans; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions