title: PrP glycoforms are associated in a strain-specific ratio in native PrPSc.
abstract: Prion diseases involve conversion of host-encoded cellular prion protein (PrPC) to a disease-related isoform (PrPSc). Using recombinant human beta-PrP, a panel of monoclonal antibodies was produced that efficiently immunoprecipitated native PrPSc and recognized epitopes between residues 93-105, indicating for the first time that this region is exposed in both human vCJD and mouse RML prions. In contrast, monoclonal antibodies raised to human alpha-PrP were more efficient in immunoprecipitating PrPC than PrPSc, and some of them could also distinguish between different PrP glycoforms. Using these monoclonal antibodies, the physical association of PrP glycoforms was studied in normal brain and in the brains of humans and mice with prion disease. It was shown that while PrPC glycoforms can be selectively immunoprecipitated, the differentially glycosylated molecules of native PrPSc are closely associated and always immunoprecipitate together. Furthermore, the ratio of glycoforms comprising immunoprecipitated native PrPSc from diverse prion strains was similar to those observed on denaturing Western blots. These studies are consistent with the view that the proportion of each glycoform incorporated into PrPSc is probably controlled in a strain-specific manner and that each PrPSc particle contains a mixture of glycoforms.
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Khalili-Shirazi
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Linda
authors.initials: L
authors.lastname: Summers
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Linehan
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Gary
authors.initials: G
authors.lastname: Mallinson
authors.affiliation: International Blood Group Reference Laboratory, Southmead Road, Bristol BS10 5NO, UK.
authors.forename: David
authors.initials: D
authors.lastname: Anstee
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Simon
authors.initials: S
authors.lastname: Hawke
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: Graham S
authors.initials: GS
authors.lastname: Jackson
authors.affiliation: MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: John
authors.initials: J
authors.lastname: Collinge
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D007562:Creutzfeldt-Jakob Syndrome / Q000276:immunology / Q000378:metabolism; D018604:Epitope Mapping; D006031:Glycosylation; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D051379:Mice; D018621:PrPC Proteins / Q000276:immunology; D018620:PrPSc Proteins / Q000737:chemistry / Q000276:immunology*; D017096:Prion Diseases / Q000276:immunology* / Q000378:metabolism; D011328:Prions / Q000276:immunology*; D020033:Protein Isoforms / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D012608:Scrapie / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; D020033:Protein Isoforms; D011994:Recombinant Proteins