NR AXLL
AU Eiden,M.; Kupfer,L.; Proft,J.; Franz,M.; Ortega Soto,E.; Groschup,M.H.
TI In-Vitro Studies on the Molecular Mechanisms of the Abnormal Prion Protein Formation
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.60
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Prion diseases are characterised by the conversion of cellular prion protein, PrPc into its pathogenic form, termed PrPsc. Compared to PrPc, PrPsc carries a partial resistance to proteinase K and forms fibrils. In a cell-free assay the conversion reaction can be imitated by incubating highly purified PrPc molecules together with PrPsc seeds in an appropriate conversion buffer. Under these conditions we can show, that PrPsc itself can induce the conversion of PrPc into a partially proteinase K resistant form, termed PrPres. Newly converted PrPres can be detected by antibodies that discriminate between PrPres and PrPsc. Bacterially expressed PrPc was used as substrate and mouse passaged scrapie and BSE strains/isolates as seeds. Even differences in the molecular masses of PrPres (after PK treatment) of mouse passaged BSE and scrapie strains (Me7, 22A, 87V, BSE/Bl6) are transmitted to the newly formed PrPres fragments which illustrates the efficacy of PrPsc to transduce its specific conformation even in this cell-free environment. Double incubation of a mouse passaged scrapie and BSE strain/isolates resulted in two PrPres fragments which were distinct in their molecular masses. Moreover, the yield of newly converted PrPres following coincubation, was higher in comparison to the individual incubation reactions indicating a synergistic interaction between the two prion strains respectively. In another set of experiments it was shown that even single or few amino acid substitutions in the prion protein have a major effect on its convertability. The cell-free assay can therefore also be used to elucidate the influence of amino acid polymorphisms on the PrP conversion process.
AD M. Eiden, L. Kupfer, J. Proft, M. Franz, E. Ortega Soto, M.H. Groschup, Friedrich-Löffler-Institut, Institut for Novel and Emerging Infectious Diseases, Germany
SP englisch
PO Schottland
EA pdf-Datei und Poster (Autorenliste um J. Proft reduziert; Titel: Molecular mechanisms of prion conversion in-vitro)