NR AOSC
AU Elfrink,K.; Schell,J.
TI Structural transitions of PrPc purified from CHO-cells in vitro
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-75
PT Konferenz-Poster
AB
The fundamental event in prion formation is the conversion of the alpha-helical, protease sensitive and non-infectious form of the prion protein (PrPc) into the insoluble, protease resistant, predominantly beta-sheeted and infectious form (PrPsc).
As described earlier1 the conversion from the alpha-helical to the beta-sheeted state can be induced by varying the SDS-concentration between 0.08% and 0.01%, respectively. So far, however, systematic experiments were carried out with recPrP, which was expressed in E. coli and thereby missed the PrPc-typical posttranslational modifications.
In the present study the in vitro conversion of PrPc purified from transgenic CHO-cells2 is analysed. These PrP-samples are glycosylated and carry a glyco-lipid-anchor similar to PrPc from hamster brain, and had never been in contact with infectivity. The purification protocol results in PrP-samples in a yield and purity grade sufficient for spectroscopic analysis. It was possible to induce beta-sheet structure, aggregation and partial PK-resistance with the specific truncation of CHO-PrPc in vitro.
In summary, it was shown that the conformational transitions, which have been discussed otherwise as specific for PrPsc-formation, can be induced in PrPc from CHO-cells, which have never been exposed to infectivity.
1 Post, K. et al. Biol. Chem. 379 (11): 1307-1317. 1998
2 Blochberger, T.C. et al. Prot. Eng. 10 (12): 1465-1473. 1997
AD K. Elfrink, J. Schell, Heinrich Heine Universität Düsseldorf Germany
SP englisch
PO Deutschland