NR AXIW
AU Bartelt-Hunt,S.L.; Saunders,S.E.; Bartz,J.C.
TI Environmental Degradation of the Prion Protein and Impacts of Prion Conformation on Environmental Behavior
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Epidemiology, Risk Assessment and Transmission P04.58
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: Recent evidence suggesting an environmental transmission pathway for chronic wasting disease (CWD) has led to interest in the environmental fate and transport of prions. In spite of this, there is limited information available about the behavior of prions in the environment.
Aims and Objectives: In this study, we investigate the environmental degradation of the prion specific isoform of the prion protein, PrPsc, to determine the most environmentally-relevant form of the protein. Soil sorption experiments were performed to investigate the impact of prion conformation on environmental behavior.
Methods: Prion-infected brain homogenate (BH) was added to a series of 200-µl tubes and was incubated at either 22° or 37°. At pre-determined time points (0, 1, 2, 7, and 35 d), tubes were removed from the incubator and analyzed by Western blot using 3F4, 8B4, and POM19 antibodies. For each experiment, a prion-infected BH (either hamster strain HY-TME or elk CWD) and an uninfected BH were used. Experiments were performed in triplicate and were conducted with pH 4, 7, and 10 to evaluate the influence of solution pH on degradation. Tests were performed to investigate sorption of full-length PrPsc and N-terminal truncated PrPsc onto montmorillonite clay and silicon dioxide microparticles. PrPsc was added to tubes containing sorbent suspended in 10 mM NaCl and were equilibrated by rotating at ambient temperature for 2 hours. Unbound PrPsc was separated from bound PrPsc by centrifugation and analyzed by Western blot.
Results: Results suggest that agents capable of PrPsc degradation exist in BH and that PrPsc degradation is evident within 24 to 48 hours. The C-terminus of PrPsc was detected at times up to 35 d at 22°, but was significantly degraded by 7 d at 37°. Data for sorption of full-length and degraded PrPsc onto montmorillonite and silicon dioxide microparticles will also be presented.
Discussion: These results indicate that N-terminal truncated PrPsc is the most environmentally-relevant form of the prion protein. PrPsc will be degraded in the environment, resulting in potentially significant conformational changes, which may impact sorption behavior. These results highlight the importance of using the most environmentally-relevant form of PrPs
AD S. Bartelt-Hunt, S. Saunders, University of Nebraska-Lincoln, USA; J. Bartz, Creighton University, USA
SP englisch
PO Schottland
EA pdf-Datei und Poster (Postertitel: Environmental Degradation of the Prion Protein)