Conformational conversion of the cellular PrPC protein to PrPSc is a central aspect of the prion diseases, but how PrP initially converts to this conformation remains a mystery. Here we show that PrP expressed in the yeast cytoplasm, instead of the endoplasmic reticulum, acquires the characteristics of PrPSc, namely detergent insolubility and a distinct pattern of protease resistance. Neuroblastoma cells cultured under reducing, glycosylation-inhibiting conditions produce PrP with the same characteristics. We therefore describe what is, to our knowledge, the first conversion of full-length PrP in a heterologous system, show the importance of reducing and deglycosylation conditions in PrP conformational transitions, and suggest a model for initiating events in sporadic and inherited prion diseases.
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We thank B. Caughey and G. Raymont for providing PrPSc, neuroblastoma cells, epitope-specific antibodies and advice; R. Kascsak for 3F4 antibody; D. Harris for mouse PrP with the 3F4 epitope; D. Ng for pDN182 plasmid; K. Runge forYEpFAT4 plasmid; and members of the Lindquist laboratory for comments. This work was supported by the NIH and the Howard Hughes Medical Institute.
Correspondence and requests for materials should be addressed to S.L.
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Ma, J., Lindquist, S. De novo generation of a PrPSc-like conformation in living cells. Nat Cell Biol 1, 358–361 (1999). https://doi.org/10.1038/14053
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