Semisynthetic murine prion protein equipped with a GPI anchor mimic incorporates into cellular membranes.
Conversion of cellular prion protein (PrP(C)) into the pathological conformer (PrP(Sc)) has been studied extensively by using recombinantly expressed PrP (rPrP). However, due to inherent difficulties of expressing and purifying posttranslationally modified rPrP variants, only a limited amount of data is available for membrane-associated PrP and its behavior in vitro and in vivo. Here, we present an alternative route to access lipidated mouse rPrP (rPrP(Palm)) via two semisynthetic strategies. These rPrP variants studied by a variety of in vitro methods exhibited a high affinity for liposomes and a lower tendency for aggregation than rPrP. In vivo studies demonstrated that double-lipidated rPrP is efficiently taken up into the membranes of mouse neuronal and human epithelial kidney cells. These latter results enable experiments on the cellular level to elucidate the mechanism and site of PrP-PrP(Sc) conversion.
Olschewski
Diana
D
Max-Planck Institute of Molecular Physiology, Department of Physical Biochemistry, Otto-Hahn-Str. 11, 44227 Dortmund, Germany.
Seidel
Ralf
R
Miesbauer
Margit
M
Rambold
Angelika S
AS
Oesterhelt
Dieter
D
Winklhofer
Konstanze F
KF
Tatzelt
Jörg
J
Engelhard
Martin
M
Becker
Christian F W
CF
D000818:Animals; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004847:Epithelial Cells / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000138:chemical synthesis*; D006801:Humans; D007668:Kidney / Q000166:cytology; D008081:Liposomes / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D018716:Molecular Mimicry; D009474:Neurons / Q000378:metabolism; D018621:PrPC Proteins / Q000737:chemistry; D018620:PrPSc Proteins / Q000138:chemical synthesis* / Q000493:pharmacokinetics; D011993:Recombinant Fusion Proteins / Q000235:genetics
D017261:Glycosylphosphatidylinositols; D008081:Liposomes; D008565:Membrane Proteins; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011993:Recombinant Fusion Proteins