NR ATVF
AU Yuan,F.F.; Biffin,S.; Brazier,M.W.; Suarez,M.; Cappai,R.; Hill,A.F.; Collins,S.J.; Sullivan,J.S.; Middleton,D.; Multhaup,G.; Geczy,A.F.; Masters,C.L.
TI Detection of prion epitopes on PrP and PrP of transmissible spongiform encephalopathies using specific monoclonal antibodies to PrP
QU Immunology and Cell Biology 2005 Dec; 83(6): 632-7
PT journal article
AB Amino acid residues 90-120 of the prion protein (PrP) are likely to be critical for the conversion of PrPc to PrPsc in the transmissible spongiform encephalopathies. We raised 10 monoclonal antibodies against the 90-120 amino acid region, mapped the epitope specificity of these anti-PrP antibodies, and investigated the expression of epitopes recognized by the antibodies in both PrPc and PrPsc. Four out of five of the anti-PrP antibodies raised in a prion knockout mouse immunized with the linear peptide of PrP90-120 could detect PrPsc in 'native' and denatured forms and PrPc in normal cells, as well as recognize epitopes within PrP93-112 residues. In contrast, the other six anti-PrP reagents, including five raised from the two knockout mice immunized with conformationally modified PrP90-120 peptide, could detect PrPc and recognize epitopes within PrP93-107 residues. Four of these reagents could also detect denatured PrPsc on western blots but not PrPsc plaques in brain tissue. The results indicate that residues PrP93-102 are exposed in PrPc but are buried upon conversion to the PrPsc isoform. Furthermore, PrP103-107 residues are partially buried in PrPsc while only the PrP107-112 epitope remains exposed, suggesting that the region PrP93-112 undergoes conformational changes during its conversion to PrPsc.
MH Animals; Antibodies, Monoclonal/*immunology; Antibody Specificity; Epitopes/chemistry/genetics/*immunology; Humans; Immunohistochemistry; Mice; PrPc Proteins/chemistry/genetics/*immunology; PrPsc Proteins/chemistry/genetics/*immunology; Prion Diseases/genetics/*immunology/*metabolism
AD Australian Red Cross Blood Service-Endeavour, Sydney, New South Wales, Australia. fyuan@arcbs.redcross.org.au
SP englisch
PO Australien