NR ATIW
AU Noinville,S.; Rigou,P.; Vasina,E.N.; Revault,M.; Abadie,J.; Quenet,Y.; Rezaei,H.; Dejardin,P.; Grosclaude,J.; Quiquampoix,H.
TI Mechanisms of interaction of PrPrec with the surfaces of soil components
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Decontamination DEC-03
PT Konferenz-Poster
AB
The fate of prions in soil is a matter of concern for both animal and human health. Assessment of dissemination risk requires the understanding of the mechanisms of interaction between prions and soil solid surfaces. PrPrec (alpha or beta structured) was used as a representative of prion entity. Montmorillonite (a clay mineral), mica and amorphous silica were chosen as models of soil surfaces.
The structure of the adsorbed protein was examined by FTIR spectroscopy. The organisation of the protein layer was determined by NMR spectroscopy. The amount of PrPrec adsorbed was quantified by NMR, FTIR, radiolabeling (125I) and depletion approaches (Western blot, ELISA). Desorption was measured in stationary or laminar flow conditions.
A highly significant finding regarding adsorption mechanisms was the observed correlation between the PrPrec structural changes and the irreversibility of adsorption. Two types of interaction were identified:
(i) With montmorillonite, alpha to beta conversion occurred. Very stringent conditions (extreme pH, detergents, high ionic strength) did not achieve desorption and competition with other proteins (animal serum) did not alter the level of PrPrec adsorption, either on montmorillonite or mica. Comparison between full length and N-truncated protein behaviour, suggested combined hydrophobic and electrostatic effects associated with the N-terminal part.
(ii) On amorphous silica, no structural conversion occurred and the adsorption was completely reversible.
An electroelution process was devised allowing efficient removal of PrPrec from montmorillonite and applicable to crude soil samples. It might open the way to quantitative and sensitive detection of prions in soil.
Considering the high efficiency of PrPrec retention by montmorillonite, trapping of infectivity is currently under investigation.
This study gives novel information on PrPrec structure in contact with surfaces and suggests practical approaches for decontamination.
IN Im Gegensatz zu Kieselerde (silica) binden die Lehmart Montmorillonit ((Na,Ca)0,3(Al,Mg)2Si4O10(OH)2 * 4H2O) und Glimmer (mica) Prionprotein derart fest, dass es es selbst mit extremen pH-Werten, Detergentien und hoher Ionenstärke nicht wieder herunter bekommt. Herauslösen konnten die Autoren das Prionprotein aus dem Tonmineral mittels Elektroelution. An Montmorillonit kam es außerdem zu Umfaltungen mit einer Zunahme des ß-Faltblattcharakters aus Kosten des Alphahelixanteil an den Sekundärstrukturen.
AD S.Noinville, M.Revault, CNRS, Laboratoire de Dynamique, Interactions et Réactivité, Thiais, France; P.Rigou, Y.Quenet, H.Rezaei, J.Grosclaude, INRA, Unité de Virologie et Immunologie Moléculaires, Jouy-en-Josas, France; E.N.Vasina, P.Déjardin, CNRS, Institut Européen des Membranes, Montpellier, France; J.Abadie, H.Quiquampoix, INRA, UMR Rhizosphère & Symbiose, Montpellier, France
SP englisch
PO Deutschland
EA
Bild 1, Bild 2, Bild 3, Bild 4, Verbund Soil, Methoden-Poster Bild 1, Methoden-Poster Bild 2, Methoden-Poster Bild 3, Methoden-Poster Bild 4
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