Strain-specific propagation of PrP(Sc) properties into baculovirus-expressed hamster PrP(C).
The conversion of the cellular isoform of the prion protein (PrP(C)) to the abnormal disease-associated isoform (PrP(Sc)) has been simulated in cell-free conversion reactions in which PrP(Sc)-enriched preparations induce the conformational transition of PrP(C) into protease-resistant PrP (PrP-res). We explored the utility of recombinant hamster (Ha)PrP(C) purified from baculovirus-infected insect cells (bacHaPrP(C)) as a replacement for mammalian-derived HaPrP(C) in the conversion reactions. Protease-resistant recombinant HaPrP was generated after incubation of (35)S-bacHaPrP(C) with PrP(Sc)-enriched preparations. Moreover strain-specific PrP-res was also reproduced using insect-cell derived HaPrP(C) and PrP(Sc) from two different strains of hamster-adapted transmissible mink encephalopathy, designated hyper (HY) and drowsy (DY). Two strain-mediated properties were tested: (i) molecular mass of the protease-digested products and (ii) relative resistance to proteinase K (PK) digestion. Similar to in vivo generation of PrP(HY) and PrP(DY), the converted products selectively reproduced both characteristics, with the DY conversion product being smaller in size and less resistant to PK digestion than the HY product. These data demonstrate that non-mammalian sources of recombinant HaPrP can be converted into PK-resistant form and that strain-mediated properties can be transmitted into the newly formed PrP-res.
Iniguez
Volga
V
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA1.
McKenzie
Debbie
D
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA1.
Mirwald
Jean
J
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA1.
Aiken
Judd
J
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA1.
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism*; D006224:Cricetinae; D004195:Disease Models, Animal; D019286:Endopeptidase K / Q000378:metabolism; D018621:PrPC Proteins / Q000378:metabolism*; D018620:PrPSc Proteins / Q000378:metabolism*; D011328:Prions / Q000472:pathogenicity; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D013045:Species Specificity; D018411:Spodoptera; D014776:Virus Cultivation
D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; D011994:Recombinant Proteins; D019286:Endopeptidase K