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@ARTICLE{Sthr:15094,
author = {Stöhr, J. and Elfrink, K. and Weinmann, N. and Wille, H.
and Willbold, D. and Birkmann, E. and Riesner, D.},
title = {{I}n vitro conversion and seeded fibrillization of
posttranslationally modified prion protein},
journal = {Biological chemistry},
volume = {392},
issn = {1431-6730},
address = {Berlin [u.a.]},
publisher = {de Gruyter},
reportid = {PreJuSER-15094},
pages = {415 - 421},
year = {2011},
note = {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>.},
abstract = {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.},
keywords = {Animals / Blotting, Western / CHO Cells / Circular
Dichroism / Cricetinae / Cricetulus / Electrophoresis,
Polyacrylamide Gel / Microscopy, Electron / PrPC Proteins:
chemistry / PrPC Proteins: drug effects / PrPC Proteins:
metabolism / PrPSc Proteins: chemistry / PrPSc Proteins:
drug effects / PrPSc Proteins: metabolism / Protein
Processing, Post-Translational / Protein Structure,
Secondary / Sodium Dodecyl Sulfate: pharmacology / PrPC
Proteins (NLM Chemicals) / PrPSc Proteins (NLM Chemicals) /
Sodium Dodecyl Sulfate (NLM Chemicals) / J (WoSType)},
cin = {ICS-6},
ddc = {540},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {Funktion und Dysfunktion des Nervensystems / BioSoft:
Makromolekulare Systeme und biologische
Informationsverarbeitung},
pid = {G:(DE-Juel1)FUEK409 / G:(DE-Juel1)FUEK505},
shelfmark = {Biochemistry $\&$ Molecular Biology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:21476870},
UT = {WOS:000289343100003},
doi = {10.1515/bc.2011.048},
url = {https://juser.fz-juelich.de/record/15094},
}