TY  - JOUR
AU  - Stöhr, J.
AU  - Weinmann, N.
AU  - Wille, H.
AU  - Kaimann, K.
AU  - Nagel-Steger, L.
AU  - Birkmann, E.
AU  - Panza, G.
AU  - Prusinder, S. B.
AU  - Eigen, M.
AU  - Riesner, D.
TI  - Mechanisms of prion protein assembly into amyloid
JO  - Proceedings of the National Academy of Sciences of the United States of America
VL  - 105
SN  - 0027-8424
CY  - Washington, DC
PB  - Academy
M1  - PreJuSER-62861
SP  - 2409 - 2414
PY  - 2008
N1  - Record converted from VDB: 12.11.2012
AB  - The conversion of the alpha-helical, cellular isoform of the prion protein (PrP(C)) to the insoluble, beta-sheet-rich, infectious, disease-causing isoform (PrP(Sc)) is the key event in prion diseases. In an earlier study, several forms of PrP were converted into a fibrillar state by using an in vitro conversion system consisting of low concentrations of SDS and 250 mM NaCl. Here, we characterize the structure of the fibril precursor state, that is, the soluble state under fibrillization conditions. CD spectroscopy, analytical ultracentrifugation, and chemical cross-linking indicate that the precursor state exists in a monomer-dimer equilibrium of partially denatured, alpha-helical PrP, with a well defined contact site of the subunits in the dimer. Using fluorescence with thioflavin T, we monitored and quantitatively described the kinetics of seeded fibril formation, including dependence of the reaction on substrate and seed concentrations. Exponential, seed-enhanced growth can be achieved in homogeneous solution, which can be enhanced by sonication. From these data, we propose a mechanistic model of fibrillization, including the presence of several intermediate structures. These studies also provide a simplified amplification system for prions.
KW  - Amyloid: chemistry
KW  - Amyloid: metabolism
KW  - Amyloid: ultrastructure
KW  - Circular Dichroism
KW  - Cross-Linking Reagents: chemistry
KW  - Dimerization
KW  - Microscopy, Electron
KW  - Prions: chemistry
KW  - Prions: metabolism
KW  - Prions: ultrastructure
KW  - Ultracentrifugation
KW  - Amyloid (NLM Chemicals)
KW  - Cross-Linking Reagents (NLM Chemicals)
KW  - Prions (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:18268326
C2  - pmc:PMC2268150
UR  - <Go to ISI:>//WOS:000253469900031
DO  - DOI:10.1073/pnas.0712036105
UR  - https://juser.fz-juelich.de/record/62861
ER  -