TY  - JOUR
AU  - Stöhr, J.
AU  - Elfrink, K.
AU  - Weinmann, N.
AU  - Wille, H.
AU  - Willbold, D.
AU  - Birkmann, E.
AU  - Riesner, D.
TI  - In vitro conversion and seeded fibrillization of posttranslationally modified prion protein
JO  - Biological chemistry
VL  - 392
SN  - 1431-6730
CY  - Berlin [u.a.]
PB  - de Gruyter
M1  - PreJuSER-15094
SP  - 415 - 421
PY  - 2011
N1  - This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG), EUNetwork of Excellence (NeuroPrion) and the Praesidentenfond of the Helmholtzgemeinschaft (HGF, Virtual Institute of Structural Biology). Jan Stohr is supported by postdoctoral fellowship of the Deutsche Forschungsgemeinschaft (DFG). We thank S.B. Prusiner for critical reading and helpful discussion for this manuscript. The authors thank Ilka Ostermann for her assistance in the purification of CHO-PrP<SUP>C</SUP>.
AB  - The conversion of the cellular isoform of the prion protein (PrP(C)) into the pathologic isoform (PrP(Sc)) is the key event in prion diseases. To study the conversion process, an in vitro system based on varying the concentration of low amounts of sodium dodecyl sulfate (SDS) has been employed. In the present study, the conversion of full-length PrP(C) isolated from Chinese hamster ovary cells (CHO-PrP(C)) was examined. CHO-PrP(C) harbors native, posttranslational modifications, including the GPI anchor and two N-linked glyco-sylation sites. The properties of CHO-PrP(C) were compared with those of full-length and N-terminally truncated recombinant PrP. As shown earlier with recombinant PrP (recPrP90-231), transition from a soluble α-helical state as known for native PrP(C) into an aggregated, β-sheet-rich PrP(Sc)-like state could be induced by dilution of SDS. The aggregated state is partially proteinase K (PK)-resistant, exhibiting a cleavage site similar to that found with PrP(Sc). Compared to recPrP (90-231), fibril formation with CHO-PrP(C) requires lower SDS concentrations (0.0075%), and can be drastically accelerated by seeding with PrP(Sc) purified from brain homogenates of terminally sick hamsters. Our results show that recPrP 90-231 and CHO-PrPC behave qualitatively similar but quantitatively different. The in vivo situation can be simulated closer with CHO-PrP(C) because the specific PK cleave site could be shown and the seed-assisted fibrillization was much more efficient.
KW  - Animals
KW  - Blotting, Western
KW  - CHO Cells
KW  - Circular Dichroism
KW  - Cricetinae
KW  - Cricetulus
KW  - Electrophoresis, Polyacrylamide Gel
KW  - Microscopy, Electron
KW  - PrPC Proteins: chemistry
KW  - PrPC Proteins: drug effects
KW  - PrPC Proteins: metabolism
KW  - PrPSc Proteins: chemistry
KW  - PrPSc Proteins: drug effects
KW  - PrPSc Proteins: metabolism
KW  - Protein Processing, Post-Translational
KW  - Protein Structure, Secondary
KW  - Sodium Dodecyl Sulfate: pharmacology
KW  - PrPC Proteins (NLM Chemicals)
KW  - PrPSc Proteins (NLM Chemicals)
KW  - Sodium Dodecyl Sulfate (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:21476870
UR  - <Go to ISI:>//WOS:000289343100003
DO  - DOI:10.1515/bc.2011.048
UR  - https://juser.fz-juelich.de/record/15094
ER  -