title: Mouse-adapted ovine scrapie prion strains are characterized by different conformers of PrPSc.
abstract: The agent responsible for prion disease may exist in different forms, commonly referred to as strains, with each carrying the specific information that determines its own distinct biological properties, such as incubation period and lesion profile. Biological strain typing of ovine scrapie isolates by serial passage in conventional mice has shown some diversity in ovine prion strains. However, this biological diversity remains poorly supported by biochemical prion strain typing. The protein-only hypothesis predicts that variation between different prion strains in the same host is manifest in different conformations adopted by PrPSc. Here we have investigated the molecular properties of PrPSc associated with two principal Prnp(a) mouse-adapted ovine scrapie strains, namely, RML and ME7, in order to establish biochemical prion strain typing strategies that may subsequently be used to discriminate field cases of mouse-passaged ovine scrapie isolates. We used a conformation-dependent immunoassay and a conformational stability assay, together with Western blot analysis, to demonstrate that RML and ME7 PrPSc proteins show distinct biochemical and physicochemical properties. Although RML and ME7 PrPSc proteins showed similar resistance to proteolytic digestion, they differed in their glycoform profiles and levels of proteinase K (PK)-sensitive and PK-resistant isoforms. In addition, the PK-resistant core (PrP27-30) of ME7 was conformationally more stable following exposure to guanidine hydrochloride or Sarkosyl than was RML PrP27-30. Our data show that mouse-adapted ovine scrapie strains can be discriminated by their distinct conformers of PrPSc, which provides a basis to investigate their diversity at the molecular level.
authors.affiliation: Department of Veterinary Medicine, University of Cambridge, Cambridge, UK.
authors.forename: Alana M
authors.initials: AM
authors.lastname: Thackray
authors.forename: Lee
authors.initials: L
authors.lastname: Hopkins
authors.forename: Michael A
authors.initials: MA
authors.lastname: Klein
authors.forename: Raymond
authors.initials: R
authors.lastname: Bujdoso
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D019286:Endopeptidase K / Q000737:chemistry; D051379:Mice; D008817:Mice, Mutant Strains; D018622:PrP 27-30 Protein / Q000032:analysis / Q000737:chemistry*; D018620:PrPSc Proteins / Q000032:analysis / Q000737:chemistry*; D011487:Protein Conformation; D012608:Scrapie / Q000378:metabolism*
chemical_list: D018620:PrPSc Proteins; D018622:PrP 27-30 Protein; D019286:Endopeptidase K