title: Heparan sulfate is a cellular receptor for purified infectious prions.
abstract: Prions replicate in the host cell by the self-propagating refolding of the normal cell surface protein, PrP(C), into a beta-sheet-rich conformer, PrP(Sc). Exposure of cells to prion-infected material and subsequent endocytosis can sometimes result in the establishment of an infected culture. However, the relevant cell surface receptors have remained unknown. We have previously shown that cellular heparan sulfates (HS) are involved in the ongoing formation of scrapie prion protein (PrP(Sc)) in chronically infected cells. Here we studied the initial steps in the internalization of prions and in the infection of cells. Purified prion "rods" are arguably the purest prion preparation available. The only proteinaceous component of rods is PrP(Sc). Mouse neuroblastoma N2a, hypothalamus GT1-1, and Chinese hamster ovary cells efficiently bound both hamster and mouse prion rods (at 4 degrees C) and internalized them (at 37 degrees C). Treating cells with bacterial heparinase III or chlorate (a general inhibitor of sulfation) strongly reduced both binding and uptake of rods, whereas chondroitinase ABC was inactive. These results suggested that the cell surface receptor of prion rods involves sulfated HS chains. Sulfated glycans inhibited both binding and uptake of rods, probably by competing with the binding of rods to cellular HS. Treatments that prevented endocytosis of rods also prevented the de novo infection of GT1-1 cells when applied during their initial exposure to prions. These results indicate that HS are an essential part of the cellular receptor used both for prion uptake and for cell infection. Cellular HS thus play a dual role in prion propagation, both as a cofactor for PrP(Sc) synthesis and as a receptor for productive prion uptake.
authors.affiliation: Department of Molecular Biology, The Hebrew University-Hadassah Medical School, and Department of Oncology, Hadassah University Hospital, Jerusalem 91120, Israel.
authors.forename: Lior
authors.initials: L
authors.lastname: Horonchik
authors.forename: Salit
authors.initials: S
authors.lastname: Tzaban
authors.forename: Olga
authors.initials: O
authors.lastname: Ben-Zaken
authors.forename: Yifat
authors.initials: Y
authors.lastname: Yedidia
authors.forename: Alex
authors.initials: A
authors.lastname: Rouvinski
authors.forename: Dulce
authors.initials: D
authors.lastname: Papy-Garcia
authors.forename: Denis
authors.initials: D
authors.lastname: Barritault
authors.forename: Israel
authors.initials: I
authors.lastname: Vlodavsky
authors.forename: Albert
authors.initials: A
authors.lastname: Taraboulos
mesh_terms: D000818:Animals; D000838:Anions; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D045744:Cell Line, Tumor; D002704:Chlorates / Q000737:chemistry; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis; D006497:Heparan Sulfate / Q000737:chemistry* / Q000502:physiology*; D008647:Mesocricetus; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D011133:Polysaccharide-Lyases / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D011328:Prions / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms; D017433:Protein Structure, Secondary; D013696:Temperature
chemical_list: D000838:Anions; D002704:Chlorates; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D011134:Polysaccharides; D011328:Prions; D020033:Protein Isoforms; C021349:heparitinsulfate lyase