title: Influence of water, fat, and glycerol on the mechanism of thermal prion inactivation.
abstract: Extending the recent analysis of the safety of industrial bovine fat-derived products for human consumption (Müller, H., Stitz, L., and Riesner, D. (2006) Eur. J. Lip. Sci. Technol. 108, 812-826), we investigated systematically the effects of fat, fatty acids, and glycerol on the heat destruction of prions. Prion destruction was qualitatively and quantitatively evaluated in PrP 27-30, or prion rods, by the inactivation of infectivity as well as by the degradation of the polypeptide backbone. Under all conditions analyzed, inactivation of prion infectivity was achieved more efficiently than backbone degradation by several orders of magnitude. The presence of fat enhanced prion inactivation and offers a mild treatment for prion decontamination. In contrast, the presence of fat, fatty acids, and especially glycerol protected the PrP 27-30 backbone against heat-induced degradation. Glycerol also protected against heat-induced inactivation of prion infectivity. A phase distribution analysis demonstrated that prions migrated to the interphase of a fat/water mixture at room temperature and accumulated in the water phase at higher temperatures. In a systematic study of the mechanism of prion destruction, we found an intermediate structure of PrP that has fewer fibrils in beta-sheet formation, lower resistance to protease digestion, greater aggregation, and reduced solubility compared with PrP 27-30 but retains residual infectivity. These findings suggest that prion infectivity depends on beta-sheet-rich fibrillar structure and that inactivation proceeds in a stepwise manner, which explains the tailing effect frequently observed during inactivation.
authors: Institut für Physikalische Biologie, Heinrich Heine-Universität Düsseldorf, D-40225 Düsseldorf, Germany.
authors: Henrik
authors: H
authors: Müller
authors: Lothar
authors: L
authors: Stitz
authors: Holger
authors: H
authors: Wille
authors: Stanley B
authors: SB
authors: Prusiner
authors: Detlev
authors: D
authors: Riesner
mesh_terms: D000818:Animals; D002417:Cattle; D005223:Fats / Q000737:chemistry*; D005990:Glycerol / Q000737:chemistry*; D006358:Hot Temperature; D006801:Humans; D044367:Phase Transition; D018622:PrP 27-30 Protein / Q000737:chemistry* / Q000472:pathogenicity; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D014867:Water / Q000737:chemistry*
chemical_list: D005223:Fats; D014867:Water; D018622:PrP 27-30 Protein; D005990:Glycerol