title: Beta-PrP form of human prion protein stimulates production of monoclonal antibodies to epitope 91-110 that recognise native PrPSc.
abstract: Prion diseases are associated with accumulation of strain-dependent biochemically distinct, disease-related isoforms (PrP(Sc)) of host-encoded prion protein (PrP(C)). PrP(Sc) is characterised by increased beta-sheet content, detergent insolubility and protease resistance. Recombinant alpha-PrP adopts a PrP(C)-like conformation, while beta-PrP conformationally resembles PrP(Sc), to these we raised 81 monoclonal antibodies in Prnp(0/0) mice. The N-terminal residues 91-110 are highly immunogenic in beta-PrP-immunised mice and of (17/41) anti-beta-PrP antibodies that could be epitope-mapped, approximately 70%, recognised this segment. In contrast, only 3/40 anti-alpha-PrP antibodies could be mapped and none interacted with this region, instead recognising residues 131-150, 141-160 and 171-190. Native PrP(C) was recognised by both antibody groups, but only anti-beta-PrP antibodies directed to 91-110 residues recognised native PrP(Sc) with high affinity, where in addition, species heterogeneity was also evident. Within the six anti-beta-PrP antibodies studied, they all recognised PK-treated native human and mouse PrP(Sc), four failed to recognise PK-treated native bovine PrP(Sc), one of which also did not recognise native PK-treated ovine PrP(Sc), showing the epitope becomes exposed on unfolding and disaggregation. These results demonstrate strain-dependent variations in chain conformation and packing within the 91-110 region of PrP(Sc).
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Diseases, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Khalili-Shirazi
authors.forename: Maria
authors.initials: M
authors.lastname: Kaisar
authors.forename: Gary
authors.initials: G
authors.lastname: Mallinson
authors.forename: Samantha
authors.initials: S
authors.lastname: Jones
authors.forename: Daljit
authors.initials: D
authors.lastname: Bhelt
authors.forename: Carol
authors.initials: C
authors.lastname: Fraser
authors.forename: Anthony R
authors.initials: AR
authors.lastname: Clarke
authors.forename: Simon H
authors.initials: SH
authors.lastname: Hawke
authors.forename: Graham S
authors.initials: GS
authors.lastname: Jackson
authors.forename: John
authors.initials: J
authors.lastname: Collinge
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D018604:Epitope Mapping; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D018621:PrPC Proteins / Q000737:chemistry / Q000276:immunology*; D018620:PrPSc Proteins / Q000737:chemistry / Q000276:immunology*; D011487:Protein Conformation; D012756:Sheep; D013045:Species Specificity
chemical_list: D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D018621:PrPC Proteins; D018620:PrPSc Proteins