Non-infectious aggregates of the prion protein react with several PrPSc-directed antibodies.
The key event in the pathogenesis of prion diseases is the conformational conversion of the normal prion protein (PrP) (PrP(C)) into an infectious, aggregated isoform (PrP(Sc)) that has a high content of beta-sheet. Historically, a great deal of effort has been devoted to developing antibodies that specifically recognize PrP(Sc) but not PrP(C), as such antibodies would have enormous diagnostic and experimental value. A mouse monoclonal IgM antibody (designated 15B3) and three PrP motif-grafted monoclonal antibodies (referred to as IgG 19-33, 89-112, and 136-158) have been previously reported to react specifically with infectious PrP(Sc) but not PrP(C). In this study, we extend the characterization of these four antibodies by testing their ability to immunoprecipitate and immunostain infectious and non-infectious aggregates of wild-type, mutant, and recombinant PrP. We find that 15B3 as well as the motif-grafted antibodies recognize multiple types of aggregated PrP, both infectious and non-infectious, including forms found in brain, in transfected cells, and induced in vitro from purified recombinant protein. These antibodies are exquisitely selective for aggregated PrP, and do not react with soluble PrP even when present in vast excess. Our results suggest that 15B3 and the motif-grafted antibodies recognize structural features common to both infectious and non-infectious aggregates of PrP. Our study extends the utility of these antibodies for diagnostic and experimental purposes, and it provides new insight into the structural changes that accompany PrP oligomerization and prion propagation.
Biasini
Emiliano
E
Department of Cell Biology and Physiology, Washington University School of Medicine, St Louis, Missouri 63110, USA.
Seegulam
M Esa
ME
Patti
Brianna N
BN
Solforosi
Laura
L
Medrano
Andrea Z
AZ
Christensen
Heather M
HM
Senatore
Assunta
A
Chiesa
Roberto
R
Williamson
R Anthony
RA
Harris
David A
DA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D018620:PrPSc Proteins / Q000276:immunology* / Q000378:metabolism* / Q000472:pathogenicity; D011328:Prions / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology; D020033:Protein Isoforms / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018620:PrPSc Proteins; D011328:Prions; D020033:Protein Isoforms