Partial purification and evidence for multiple molecular forms of the scrapie agent.
A procedure for the partial purification of the scrapie agent from mouse spleen was developed based on its sedimentation profile. Differential centrifugation and detergent treatment with sodium deoxycholate yielded a fraction designated "P5" which was enriched for scrapie infectivity approximately 20-fold with respect to cellular protein. The P5 fraction was devoid of cellular membranes but heavily contaminated with ribosomes as judged by electron microscopy. On centrifugation of the fraction P5 to near equilibrium in a sucrose gradient scrapie infectivity was distributed over a range of densities from 1.08 to 1.30 g/cm3. Parallel rate-zonal analysis showed that the infectivity was distributed over a range of particle sizes with s20.w values from approximately 40 S to greater than 500 S. Incubation of P5 at 37 or 80 degrees C, under conditions that disrupt ribosomes, dramatically altered the rate-zonal gradient profile of the agent. Under these conditions, the agent sedimented as particles with s20.w greater than 500 S. The apparent heterogeneity of the scrapie agent with respect to both size and density and its ability to shift from one form to another suggest that the agent may contain hydrophobic domains on its surface.
D000818:Animals; D002499:Centrifugation, Density Gradient; D004279:DNA, Viral / Q000032:analysis; D051379:Mice; D008854:Microscopy, Electron; D011328:Prions / Q000032:analysis*; D012367:RNA, Viral / Q000032:analysis; D013154:Spleen / Q000382:microbiology*; D014764:Viral Proteins / Q000032:analysis
D004279:DNA, Viral; D011328:Prions; D012367:RNA, Viral; D014764:Viral Proteins