NR AXUB
AU Panza,G.; Weinmann,N.; Stöhr,J.; Birkmann,E.; Willbold,D.; Riesner,D.
TI Seeded Fibrillization of Recombinant Bovine Prion with BSE-prions
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.38
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Conversion of the cellular isoform of the prion protein (PrPc) to the infectious isoform of PrP (PrPsc) plays a key role in the development of prion diseases. To simulate the structural transition in vitro we developed a solution system of well balanced concentrations of SDS and NaCl. Under these conditions an intermediate, partially denatured state of PrP is soluble for weeks but forms fibrils afterward (Leffers et al. 2005, Stöhr et al. manuscript submitted ).
In the present work we show for the first time the conversion of bov recPrP (25-241) into amyloid as assayed by Thioflavin T (ThT)-fluorescence-assay. Furthermore we extended the study in respect to seeded fibrilogenesis of bov recPrP. This study is based on a seeded-conversion assay as established in our group using SHa recPrP and hamster scrapie prions (Weinmann et al. manuscript in preparation). In the present study minute amounts of purified bovine BSE-prions prepared without PK-digestion serve as effctive seeds for the conversion of bov recPrP (25-241) into an fibrillar, amyloid state as proven by ThT-fluorescence-assay. This conversion is specific for the seed, since it occurs within hours, whereas bov recPrP in the absence of seeds is converted into fibres only after weeks of incubation. The system is similar to the seeded-fibrillization of the Soto-group (Castilla et. al 2005) but clearly different in two aspects, first that it is purely in vitro i.e. without any cellular compounds and second it avoids incubation-sonification cycles by strong homogenous stirring.G.
AD G. Panza, N. Weinmann, J. Stöhr, E. Birkmann, D. Willbold, D. Riesner, Heinrich-Heine-Universität, Institut für Physikalische Biologie, Germany
SP englisch
PO Schottland