PrP glycoforms are associated in a strain-specific ratio in native PrPSc.
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.
Khalili-Shirazi
Azadeh
A
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
Summers
Linda
L
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
Linehan
Jacqueline
J
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
Mallinson
Gary
G
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
Anstee
David
D
International Blood Group Reference Laboratory, Southmead Road, Bristol BS10 5NO, UK.
Hawke
Simon
S
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
Jackson
Graham S
GS
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
Collinge
John
J
MRC Prion Unit, Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
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
D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; D020033:Protein Isoforms; D011994:Recombinant Proteins