Methods for Studying Prion Protein (PrP) Metabolism and the Formation of Protease-Resistant PrP in Cell Culture and Cell-Free Systems

The pathogenesis of scrapie and other transmissible spongiform encephalopathies (TSEs) appears to be based on the posttranslational conversion of the host’s protease-sensitive prion protein (PrP-sen or PrP^c ) to abnormal protease-resistant forms (PrP-res or PrP^Sc ). In vitro studies using both scrapie-infected tissue culture cells and cell-free reactions have been used effectively to help define the cellular and molecular details of this process and how it might be inhibited. Here we discuss experimental approaches that we have used in these studies