title: Some physico-chemical parameters that influence proteinase K resistance and the infectivity of PrPSc after high pressure treatment.
abstract: Crude brain homogenates of terminally diseased hamsters infected with the 263 K strain of scrapie (PrP Sc) were heated and/or pressurized at 800 MPa at 60 degrees C for different times (a few seconds or 5, 30, 120 min) in phosphate-buffered saline (PBS) of different pH and concentration. Prion proteins were analyzed on immunoblots for their proteinase K (PK) resistance, and in hamster bioassays for their infectivity. Samples pressurized under initially neutral conditions and containing native PrP Sc were negative on immunoblots after PK treatment, and a 6-7 log reduction of infectious units per gram was found when the samples were pressurized in PBS of pH 7.4 for 2 h. A pressure-induced change in the protein conformation of native PrP Sc may lead to less PK resistant and less infectious prions. However, opposite results were obtained after pressurizing native infectious prions at slightly acidic pH and in PBS of higher concentration. In this case an extensive fraction of native PrP Sc remained PK resistant after pressure treatment, indicating a protective effect possibly due to induced aggregation of prion proteins in such buffers.
authors.affiliation: Federal Research Center for Nutrition and Food, Institute of Chemistry and Biology, Karlsruhe, Germany. philipp.heindl@bfe.uni-karlsruhe.de
authors.forename: P
authors.initials: P
authors.lastname: Heindl
authors.forename: A
authors.initials: A
authors.lastname: Fernández García
authors.forename: M
authors.initials: M
authors.lastname: Büttner
authors.forename: H
authors.initials: H
authors.lastname: Voigt
authors.forename: P
authors.initials: P
authors.lastname: Butz
authors.forename: B
authors.initials: B
authors.lastname: Tauscher
authors.forename: E
authors.initials: E
authors.lastname: Pfaff
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D002021:Buffers; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D006224:Cricetinae; D019286:Endopeptidase K / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006874:Hydrostatic Pressure; D018620:PrPSc Proteins / Q000737:chemistry* / Q000378:metabolism / Q000472:pathogenicity; D013997:Time Factors
chemical_list: D002021:Buffers; D018620:PrPSc Proteins; D019286:Endopeptidase K