NR AXUZ
AU Redecke,L.; Bergmann,J.; Bergmann,R.; Betzel,C.; Preddie,E.
TI Mimicking Pathological Conversion of PrP to PrPsc in Vitro
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.36
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB The pivotal event in the initiation of prion disease is the conversion of normal cellular prion protein (PrPc) into PrPsc that is believed to be made up entirely of miss-folded PrP molecules which become tightly aggregated. The consensus of scientific opinion is that only PrP molecules are involved in this process. We have discovered an alternative prion protein ("prionin") that is encoded and expressed from within the normal PrP transcription unit in all species of PrP genes. Prionins are small (39 amino acids in hamsters to 102 in mouse) proteins that are expressed in a prion diseasespecific manner and they are transmissible between species. In this study we show that prionins can convert PrP to proteinase K-resistant conformers in vitro. Nanogram quantities of a "converting factor" (cf; a sequence of amino acids in a conserved region located in the bovine, ovine and deer prionin, chemically synthesized > 95% purity) converted native PrPc from healthy humans (10 % extracts of brain), cows, sheep and full length recombinant human PrP (in PBS) to aggregates with a 27-29 kDa proteinase K-resistant core as well as truncated, recombinant human, cattle and murine PrP (in PBS) to a 22-25 kDa proteinase K-resistant conformer. Conversion started immediately upon addition of cf to the various PrP preparations and proceeded with time during incubation with gentle shaking (2.5x100 min-1) at 4 °, ambient temperature and 37°. No infectious seeding material was added to any of the incubations. Conversion was inhibited by an amphipathic peptide from a naturally occurring mammalian protein and by doxycycline. For further inhibition studies we have developed an antibody against a 4-amino acid sequence within cf. Moreover, infection and vaccination studies are on the way using hamsters to prove the converting effect of prionins in vivo.
AD L. Redecke, C. Betzel, University of Hamburg, Institute for Biochemistry and Molecular Biology, Germany; J. Bergmann, E. Preddie, Altegen, Inc., Canada; R. Bergmann, University of Hamburg, Biocenter Klein Flottbek, Germany
SP englisch
PO Schottland