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United States Department of Agriculture

Agricultural Research Service

Title: Strain-Specific Prp-Res in Vitro Amplification by the Pmca Technique

Authors
item Zukas, Audrius
item Carter, John

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: February 14, 2006
Publication Date: March 7, 2006
Citation: Zukas, A.A., Carter, J.M. 2006. Strain-specific prp-res in vitro amplification by the pmca technique. [Abstract]. Cambridge Healthtec Institute, Transmissible Spongiform Encephalopathies. Poster #12.

Interpretive Summary: We have investigated in vitro conversion of physiological prion protein to the 263K and drowsy strains of the scrapie prion protein. We used the Protein Misfolding Cyclic Amplification (PMCA) technique to convert physiological prions to scrapie prions. PMCA resulted in a significant conversion for both strains. We found that the amplified 263K strain displayed similar characteristics to that of the preamplified 263K strain. However, the amplified drowsy strain became more resistant towards enzyme degradation suggesting that it converted to a different strain.

Technical Abstract: The 263K and drowsy (DY) strains of the scrapie prion protein (PrPSC) are post-translationally derived from the physiological prion protein (PrPC) precursor. In vivo, the two strain-specific conformers can self-propagate by converting the PrPC to the PrPSC 263K and DY conformers. The 263K and DY strains have different incubation periods, clinical symptoms, biochemical, and physical properties. The two distinct protein conformations result in different gel migration patterns due to differences in protease K (PK) cleavage site. In addition, the DY strain is more susceptible towards PK digestion compared to the 263K strain. We have investigated in vitro conversion of PrPC to the 263K and DY strains using the Protein Misfolding Cyclic Amplification (PMCA) technique. PMCA resulted in a significant conversion of PrPC to PK-resistant PrP (PrP-res) for both strains. We found that the amplified 263K strain displayed similar characteristics to that of the preamplified 263K strain. However, the amplified DY strain became more PK resistant suggesting a different conformation.

Last Modified: 11/1/2014
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