PrPSc incorporation to cells requires endogenous glycosaminoglycan expression.
Many lines of evidence suggest an interaction between glycosaminoglycans (GAGs) and the PrP proteins as well as a possible role for GAGs in prion disease pathogenesis. In this work, we sought to determine whether the PrP-GAG interaction affects the incorporation of PrP(Sc) (the scrapie isoform of PrP) to normal cells. This may be the first step in prion disease pathogenesis. To this effect, we incubated proteinase K-digested hamster scrapie brain homogenates with several lines of Chinese hamster ovary (CHO) cells in the presence or absence of heparin. Our results show that over a large range of PrP(Sc) concentrations the binding of PrP(Sc) to wild type CHO cells, which do not express detectable PrP, was equivalent to the binding of PrP(Sc) to CHO cells overexpressing PrP. A significant part of PrP(Sc) binding to both lines could be inhibited by heparin. Additional evidence that PrP(Sc) binding to cells was dependent on the presence of GAGs could be concluded from the fact that the binding of PrP(Sc) to CHO cells missing GAGs on the cell surface was significantly reduced. Interestingly, preincubation of scrapie brain homogenate with heparin before intraperitoneal inoculation into normal hamsters resulted in a significant delay in prion disease manifestation.
Hijazi
Nuha
N
Department of Neurology, The Agnes Ginges Center for Human Neurogenetics, Hadassah University Hospital, Jerusalem 91120, Israel.
Kariv-Inbal
Zehavit
Z
Gasset
Maria
M
Gabizon
Ruth
R
D000818:Animals; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin / Q000737:chemistry / Q000378:metabolism; D008647:Mesocricetus; D051379:Mice; D018620:PrPSc Proteins / Q000378:metabolism*; D011328:Prions / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D012608:Scrapie / Q000378:metabolism; D012685:Sepharose / Q000737:chemistry; D013696:Temperature
D006025:Glycosaminoglycans; D018620:PrPSc Proteins; D011328:Prions; D006493:Heparin; D012685:Sepharose