NR AXUU
AU Priem,J.; Langeveld,J.; van Zijderveld,F.; Bossers,A.
TI Ovine and Bovine Species/polymorphism-specificity of the Protein Misfolding Cyclic Amplification (PMCA)
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.29
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Polymorphisms in the prion protein (host and donor PrP genotype) are known to influence TSE susceptibility and transmissibility in vivo.
These modulating effects on the underlying molecular conversion of prion proteins can be readily assessed in vitro.
The so-called VRQ allelic variant of sheep PrP is generally associated with high susceptibility to sheep scrapie.
The ARR allelic variant is in general considered to encode lowered susceptibility for scrapie, although this might be questioned for atypical and/or sheep derived BSE.
In vitro tests assessing the (potential) species/polymorphism barriers support these findings on the molecular level and they can measure virtually every species barrier.
In the past we used a Gdn-based conversion system to measure the effect of speciesand polymorphism-barriers on prion protein conversion efficiencies.
These studies revealed a clear correlation between barriers observed in vivo and the corresponding in vitro conversion efficiencies.
Here we describe the use of the PMCA technique to measure conversions efficiencies between ovine and bovine allelic variants.
The protein misfolding cyclic amplification (PMCA) is a technique allowing the in vitro generation of protease-resistant prion protein (PrPres) using only a small seed of disease-associated prion protein (PrPsc) to convert normal cellular prion protein (PrPc) in brain homogenates in contrast to Gdn-based systems.
Several incubation and sonication steps allows the diluted positive starting material to be amplified, detectable by Western blot analysis but also by bioassays.
After fine-tuning (for specificity) the PMCA conditions to ovine and bovine derived materials, we were able to measure species/polymorphism-barriers as determined before using Gdn-based conversion reactions.
Conversion specificity ("transmission profiles"), reproducibility, (in)stability of glycoprofiles, as well as the surprising "transmission profile" of sheep derived BSE, will be discussed.
AD J. Priem, J. Langeveld, F. van Zijderveld, A. Bossers, CIDC-Lelystad, Department of bacteriology and TSE's, Netherlands
SP englisch
PO Schottland
EA pdf-Datei und Poster (Autorenliste um O. Andreoletti ergänzt)