Mapping the antigenicity of copper-treated cellular prion protein with the scrapie isoform.
When recombinant and cellular prion protein (PrP(C)) binds copper, it acquires properties resembling the scrapie isoform (PrP(Sc)), namely protease resistance, detergent insolubility and increased beta sheet content. However, whether the conformations of PrP(C) induced by copper and PrP(Sc) are similar has not been studied in great detail. Here, we use a panel of seven monoclonal antibodies to decipher the epitopes on full-length mouse PrP(C) that are affected by exogenous copper, and to compare the antigenicity of the copper-treated full-length PrP(C) with the full-length PrP(Sc) present in scrapie-infected mouse brains. In the presence of copper, we found that epitopes along residues 115-130 and 153-165 become more accessible on PrP(C). These regions correspond to the two beta sheet strands in recombinant PrP and they were proposed to be important for prion conversion. However, when we compared the antibody-binding patterns between full-length PrP(C) with full-length PrP(Sc) and between copper-treated full-length PrP(C) with full-length PrP(Sc), antibody binding to residues 143-155 and 175-185 was consistently increased on PrP(Sc). Collectively, our results suggest that copper-treated full-length PrP(C) does not resemble full-length PrP(Sc), despite acquiring PrP(Sc)-like properties. In addition, since each full-length protein reacts distinctively to some of the antibodies, this binding pattern could discriminate between PrP(C) and PrP(Sc).
Wong
B-S
BS
Institute of Pathology (BRB 933), Case Western Reserve University, School of Medicine, 10900 Euclid Ave., Cleveland, Ohio 44106, USA.
Li
R
R
Sassoon
J
J
Kang
S-C
SC
Liu
T
T
Pan
T
T
Greenspan
N S
NS
Wisniewski
T
T
Brown
D R
DR
Sy
M-S
MS
D000818:Animals; D000911:Antibodies, Monoclonal; D001665:Binding Sites; D001923:Brain Chemistry; D003300:Copper / Q000494:pharmacology*; D018604:Epitope Mapping; D066298:In Vitro Techniques; D051379:Mice; D018345:Mice, Knockout; D018621:PrPC Proteins / Q000737:chemistry / Q000187:drug effects* / Q000276:immunology*; D018620:PrPSc Proteins / Q000737:chemistry / Q000276:immunology*; D011487:Protein Conformation / Q000187:drug effects; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011994:Recombinant Proteins; D003300:Copper