Generating a prion with bacterially expressed recombinant prion protein.
The prion hypothesis posits that a misfolded form of prion protein (PrP) is responsible for the infectivity of prion disease. Using recombinant murine PrP purified from Escherichia coli, we created a recombinant prion with the attributes of the pathogenic PrP isoform: aggregated, protease-resistant, and self-perpetuating. After intracerebral injection of the recombinant prion, wild-type mice developed neurological signs in approximately 130 days and reached the terminal stage of disease in approximately 150 days. Characterization of diseased mice revealed classic neuropathology of prion disease, the presence of protease-resistant PrP, and the capability of serially transmitting the disease; these findings confirmed that the mice succumbed to prion disease. Thus, as postulated by the prion hypothesis, the infectivity in mammalian prion disease results from an altered conformation of PrP.
Wang
Fei
F
Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, OH 43210, USA.
Wang
Xinhe
X
Yuan
Chong-Gang
CG
Ma
Jiyan
J
D000818:Animals; D001921:Brain / Q000473:pathology; D001923:Brain Chemistry; D002460:Cell Line; D019286:Endopeptidase K / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006031:Glycosylation; D008099:Liver / Q000737:chemistry; D051379:Mice; D009474:Neurons / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry*; D018621:PrPC Proteins / Q000737:chemistry / Q000472:pathogenicity; D018620:PrPSc Proteins / Q000032:analysis / Q000737:chemistry* / Q000472:pathogenicity*; D017096:Prion Diseases / Q000209:etiology* / Q000473:pathology; D011328:Prions / Q000737:chemistry* / Q000472:pathogenicity*; D017510:Protein Folding; D012313:RNA / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry
D010715:Phosphatidylglycerols; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; D011994:Recombinant Proteins; D012313:RNA; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; D019286:Endopeptidase K