title: Enzymatic degradation of prion protein in brain stem from infected cattle and sheep.
abstract: Prions-infectious agents involved in transmissible spongiform encephalopathies-normally survive proteolytic and mild protein-destructive processes. Using bacterial keratinase produced by Bacillus licheniformis strain PWD-1, we tested conditions to accomplish the full degradation of prion protein (PrP) in brain-stem tissue from animals with bovine spongiform encephalopathy and scrapie. The detection of PrPSc, the disease-associated isoform of PrP, in homogenates was done by Western blotting and various antibodies. The results indicated that only in the presence of detergents did heat pretreatment at >100 degrees C allow the extensive enzymatic breakdown of PrPSc to a state where it is immunochemically undetectable. Proteinase K and 2 other subtilisin proteases, but not trypsin and pepsin, were also effective. This enzymatic process could lead to the development of a method for the decontamination of medical and laboratory equipment. The ultimate effectiveness of this method of prion inactivation has to be tested in mouse bioassays.
authors.affiliation: Division of Infectious Diseases and Food Chain Quality, Institute for Animal Science and Health, Lelystad, The Netherlands. jan.langeveld@wur.nl.
authors.forename: Jan P M
authors.initials: JP
authors.lastname: Langeveld
authors.forename: Jeng-Jie
authors.initials: JJ
authors.lastname: Wang
authors.forename: Dick F M
authors.initials: DF
authors.lastname: Van de Wiel
authors.forename: Giles C
authors.initials: GC
authors.lastname: Shih
authors.forename: G Jan
authors.initials: GJ
authors.lastname: Garssen
authors.forename: Alex
authors.initials: A
authors.lastname: Bossers
authors.forename: Jason C H
authors.initials: JC
authors.lastname: Shih
mesh_terms: D000818:Animals; D001933:Brain Stem / Q000378:metabolism*; D002417:Cattle; D003666:Decontamination; D010447:Peptide Hydrolases / Q000378:metabolism; D017096:Prion Diseases / Q000378:metabolism*; D011328:Prions / Q000378:metabolism*; D011489:Protein Denaturation; D012756:Sheep; D013696:Temperature
chemical_list: D011328:Prions; D010447:Peptide Hydrolases; C035855:keratinase