Acidic pH and detergents enhance in vitro conversion of human brain PrPC to a PrPSc-like form.
In the presence of a low concentration of denaturants or detergents, acidic pH triggers a conformational transition of alpha-helices into beta-sheets in recombinant prion protein (PrP), likely mimicking some aspects of the transformation of host-encoded normal cellular PrP (PrP(C)) into its pathogenic isoform (PrP(Sc)). Here we observed the effects of acidic pH and guanidine hydrochloride (GdnHCl) on the physicochemical and structural properties of PrP(C) derived from normal human brain and determined the ability of the acid/GdnHCl-treated PrP to form a proteinase K (PK)-resistant species in the absence and presence of PrP(Sc) template. After treatment with 1.5 m GdnHCl at pH 3.5, PrP(C) from normal brain homogenates was converted into a detergent-insoluble form similar to PrP(Sc). Unlike PrP(Sc), however, the treated brain PrP(C) was protease-sensitive and retained epitope accessibility to monoclonal antibodies 3F4 and 6H4. Brain PrP(C) treated with acidic pH/GdnHCl acquired partial PK resistance upon further treatment with low concentrations of sodium dodecyl sulfate (SDS). Formation of this PrP(Sc)-like isoform was greatly enhanced by incubation with trace quantities of PrP(Sc) from Creutzfeldt-Jakob disease brain. Acid/GdnHCl-treated brain PrP may constitute a "recruitable intermediate" in PrP(Sc) formation. Further structural rearrangement seems essential for this species to acquire PK resistance, which can be promoted by the presence of a PrP(Sc) template.
Zou
Wen-Quan
WQ
Centre for Research in Neurodegenerative Diseases and Sunnybrook & Women's College Health Sciences Centre, University of Toronto, Ontario M5S 3H2, Canada.
Cashman
Neil R
NR
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001921:Brain / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D007562:Creutzfeldt-Jakob Syndrome / Q000473:pathology; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D019286:Endopeptidase K / Q000494:pharmacology; D000939:Epitopes; D019791:Guanidine / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D018621:PrPC Proteins / Q000737:chemistry / Q000378:metabolism*; D018620:PrPSc Proteins / Q000737:chemistry / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary
D000911:Antibodies, Monoclonal; D003902:Detergents; D000939:Epitopes; D018621:PrPC Proteins; D018620:PrPSc Proteins; D020033:Protein Isoforms; D019286:Endopeptidase K; D019791:Guanidine