title: Conversion of bacterially expressed recombinant prion protein.
abstract: The infectivity associated with prion disease sets it apart from a large group of late-onset neurodegenerative disorders that shares the characteristics of protein aggregation and neurodegeneration. The unconventional infectious agent, PrP(Sc), is an aberrantly folded form of the normal prion protein (PrP(C)) and the PrP(C)-to-PrP(Sc) conversion is a critical pathogenic step in prion disease. Using the Protein Misfolding Cyclic Amplification technique, we converted folded bacterially expressed recombinant PrP into a proteinase K-resistant and aggregated conformation (rPrP-res) in the presence of anionic lipid and RNA molecules. Moreover, high prion infectivity was demonstrated by intracerebral inoculation of rPrP-res into wild-type mice, which caused prion disease with a short incubation period. The establishment of the in vitro recombinant PrP conversion assay makes it feasible for us to explore the molecular basis behind the intriguing properties associated with prion infectivity.
authors.affiliation: Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Xinhe
authors.initials: X
authors.lastname: Wang
authors.forename: Jiyan
authors.initials: J
authors.lastname: Ma
mesh_terms: D000818:Animals; D001921:Brain / Q000473:pathology; D003001:Cloning, Molecular / Q000379:methods; D004195:Disease Models, Animal; D019286:Endopeptidase K / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D005260:Female; D051379:Mice; D010715:Phosphatidylglycerols / Q000737:chemistry; D018621:PrPC Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000737:chemistry*; D018620:PrPSc Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000737:chemistry*; D017096:Prion Diseases / Q000473:pathology; D017510:Protein Folding; D012313:RNA / Q000737:chemistry; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000096:biosynthesis / Q000737:chemistry*; D013010:Sonication
chemical_list: D010715:Phosphatidylglycerols; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011994:Recombinant Proteins; D012313:RNA; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; D019286:Endopeptidase K