the physical relationship between infectivity and prion protein aggregates is strain-dependent物理strain-dependent传染性和朊蛋白总量之间的关系.pdfVIP

The Physical Relationship between Infectivity and Prion Protein Aggregates Is Strain-Dependent ¨ ´ ˆ Philippe Tixador, Laetitia Herzog, Fabienne Reine, Emilie Jaumain, Jerome Chapuis, Annick Le Dur, ´ Hubert Laude*, Vincent Beringue* ´ INRA (Institut National de la Recherche Agronomique), UR892, Virologie Immunologie Moleculaires, Jouy-en-Josas, France Abstract Prions are unconventional infectious agents thought to be primarily composed of PrPSc, a multimeric misfolded conformer of the ubiquitously expressed host-encoded prion protein (PrPC). They cause fatal neurodegenerative diseases in both animals and humans. The disease phenotype is not uniform within species, and stable, self-propagating variations in PrPSc conformation could encode this ‘strain’ diversity. However, much remains to be learned about the physical relationship between the infectious agent and PrPSc aggregation state, and how this varies according to the strain. We applied a sedimentation velocity technique to a panel of natural, biologically cloned strains obtained by propagation of classical and atypical sheep scrapie and BSE infectious sources in transgenic mice expressing ovine PrP. Detergent-solubilized, infected brain homogenates were used as starting material. Solubilization conditions were optimized to separate PrPSc aggregates from PrPC. The distribution of PrPSc and infectivity in the gradient was determined by immunoblotting and mouse bioassay, respectively. As a general feature, a major proteinase K-resistant PrPSc peak was observed in the middle part of the gradient. This population approximately corresponds