NR AXTI
AU Muyrers,J.; Breil,A.; Müller-Schiffmann,A.; Leliveld,R.; Onisko,B.; Lingappa,V.; Korth,C.
TI Monoclonal Antibodies Define Conformational Heterogeneity of the Normal Prion Protein
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.157
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
The physiological function of the cellular prion protein PrPc is still unclear. Seemingly opposing functions, namely toxic or protective ones have been proposed. These effects could be explained by PrPc adopting several stable conformations which was shown in in vitro translation studies; here, PrP mRNA is translated into secretory GPIanchored SECPrP and the two transmembrane isoforms NTMPrP and CTMPrP. Due to the absence of specific ligands to these isoforms definite proof of conformational heterogeneity of PrPc in vivo has not been demonstrated.
The aim of our study was therefore to provide such ligands and use them to characterize the cell biology of these conformers.
Immunization of PrP ko mice with recombinant purified mouse PrP, and screening by immunoprecipitation of in vitro translated PrP yielded two hybrodima cell lines secreting:
a.) mAB19B10 exclusively recognizing a conformational epitope of an unglycosylated PrP subpopulation overlapping with the NTMPrP from in vitro translation studies. Immunofluorescence staining of cells by overexpressing NTMPrP-favoring PrP constructs showed distinct staining patterns defining a distinct cellular compartment for NTMPrP. Pull-down experiments with recombinant antibody fragments of these antibodies enabled us to identify a specific ligand for NTMPrP.
b.) mAB19C3 recognizing CTMPrP. Using 19C3 we were able to selectively immunoprecipitate CTMPrP from scrapie-infected, but not normal mouse brains suggesting close association of CTMPrP with prion infection and suggesting neurotoxic potential of this conformer.
We thus provide evidence that "normal" PrPc consists in at least three different conformers in vivo. PrPc's conformational heterogeneity is used to differentially carry out biological functions.
AD J. Muyrers, A. Breil, A. Müller-Schiffmann, R. Leliveld, C. Korth, University of Duesseldorf, Germany; B. Onisko, USDA, USA; V. Lingappa, University of California San Francisco, USA
SP englisch
PO Schottland
EA pdf-Datei und Poster (Posterautorenliste um A. Müller-Schiffmann und R. Leliveld verkürzt)