NR AFUI
AU Iniguez,V.; McKenzie,D.; Mirwald,J.; Aiken,J.M.
TI Strain-specific propagation of PrPsc properties into baculovirus-expressed hamster PrPc
QU Journal of General Virology 2000 Oct; 81(10): 2565-71
PT journal article
AB The conversion of the cellular isoform of the prion protein (PrPc) to the abnormal disease-associated isoform (PrPsc) has been simulated in cell-free conversion reactions in which PrPsc-enriched preparations induce the conformational transition of PrPc into protease-resistant PrP (PrPres). We explored the utility of recombinant hamster (Ha)PrPc purified from baculovirus-infected insect cells (bacHaPrPc) as a replacement for mammalian-derived HaPrPc in the conversion reactions. Protease-resistant recombinant HaPrP was generated after incubation of (35)S-bacHaPrPc with PrPsc-enriched preparations. Moreover strain-specific PrPres was also reproduced using insect-cell derived HaPrPc and PrPsc 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 PrPres.
MH Animal; Baculoviridae/*metabolism; Disease Models, Animal; Endopeptidase K/metabolism; Hamsters; PrPc Proteins/*metabolism; PrPsc Proteins/*metabolism; Prions/pathogenicity; Protein Conformation; Recombinant Proteins/metabolism; Species Specificity; Spodoptera; Virus Cultivation
AD Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison, 1656 Linden Drive, Madison, WI 53706, USA
SP englisch
PO England