Since previous studies have shown that PrP-converting - Nature由于先前的研究已经表明，PRP转换性.docVIP

Supplementary Information The most infectious prion protein particles Jay R. Silveira1, Gregory J. Raymond1, Andrew G. Hughson1, Richard E. Race1, Valerie L. Sim1, Stanley F. Hayes2, and Byron Caughey1 1Laboratory of Persistent Viral Diseases and 2Electron Microscopy Core Facility, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840, USA Table of Contents 1. Supplementary Methods 2. Supplementary Figure S1: Filtration of converting activity in SDS-treated samples of PrP-res. 3. Supplementary Figure S2: 300-kDa filtration of converting activity in alkyl sulfate-treated samples of PrP-res. 4. Supplementary Figure S3: Suspension-based conversion assay of fractionated PrP-res. 5. Supplementary Figure S4: Titration of scrapie infectivity 6. Supplementary Figure S5: PAGE analyses of SUS-treated PrP-res. 7. Supplementary Figure S6: Denaturing PAGE analyses of fractionated PrP-res. 8. Supplementary Discussion 9. Supplementary References 1. Supplementary Methods Analysis of filtered converting activity Samples of purified, PK-digested 263K PrP-res (2 mg/mL) were pelleted (20,800 x g, 20 min, 4°C), and resuspended in an equal volume of 0.25 M Tris pH 7.0. Aliquots (0.5-7.5 uL) were diluted to 0.1 mg/mL in the same buffer containing various detergents, without sonication. Samples were subjected to a 5 min freeze/thaw cycle, incubated at 37°C for one hour, and filtered by centrifugation (14,000 x g, 3 min or 500 x g, 20 min) at 25 °C through 0.2 (m or 300 kDa (~30 nm) Nanosep centrifugal devices. Filtrates of the material were analyzed by solid-phase conversion assay, and the formation of small particles that retained converting activity was assessed by the levels of converting activity in filtrates, which were expressed as a percentage of the level of converting activity detected in untreated, unfiltered PrP-res. Suspension-based PrP conversion Samples (20 (L) of fractiona