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Published on: August 8, 2017
Generating a prion with bacterially expressed recombinant prion protein
Fei Wang1, Xinhe Wang, Chong-Gang Yuan
1Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, OH 43210, USA.
Abstract:
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.
Insights
Researchers created a recombinant prion protein that caused prion disease in mice. This supports the prion hypothesis that misfolded prion protein (PrP) causes these fatal neurodegenerative conditions.
Area of Science:
- Neuroscience
- Molecular Biology
- Protein Chemistry
Background:
- Prion diseases are fatal neurodegenerative disorders.
- The prion hypothesis suggests a misfolded prion protein (PrP) isoform causes infectivity.
- Understanding PrP misfolding is crucial for disease mechanisms.
Purpose of the Study:
- To create a recombinant prion with pathogenic characteristics.
- To test if this recombinant prion can induce prion disease in vivo.
- To validate the prion hypothesis using a recombinant PrP model.
Main Methods:
- Purification of recombinant murine prion protein (PrP) from Escherichia coli.
- Generation of a recombinant prion exhibiting aggregation, protease resistance, and self-propagation.
- Intracerebral injection of recombinant prion into wild-type mice.
Main Results:
- Mice injected with recombinant prion developed neurological signs around 130 days.
- Diseased mice reached terminal stages by approximately 150 days post-injection.
- Neuropathology, protease-resistant PrP, and disease transmission confirmed prion disease.
Conclusions:
- The study successfully generated a recombinant prion capable of causing disease.
- Findings provide direct experimental support for the prion hypothesis.
- Infectivity in mammalian prion disease is attributed to an altered PrP conformation.
