NR AJSZ
AU Prusiner,S.B.; Garfin,D.E.; Cochran,S.P.; McKinley,M.P.; Groth,D.F.; Hadlow,W.J.; Race,R.E.; Eklund,C.M.
TI Experimental scrapie in the mouse: electrophoretic and sedimentation properties of the partially purified agent.
QU Journal of Neurochemistry 1980 Sep; 35(3): 574-82
PT journal article
AB Some biochemical and biophysical properties of the scrapie agent in a partially purified fraction P5 from murine spleen are described in this communication. The agent was stable in the nonionic detergents Triton X-100 and Nonidet P40 and stable in the nondenaturing, anionic detergents sodium cholate and sodium N-lauroyl sarcosinate. In contrast, sodium dodecyl sulfate (SDS) inactivated the agent at high concentrations (1% or >) when the detergent-to-protein ration approached 1.5 g SDS/g protein. The agent was resistant to inactivation by nucleases and proteases, even in the presence of 0.1% SDS. A broad peak of infectivity was exhibited in modified colloidal silica (Percoll) density gradients. Maximal titers were found at a Percoll density of 1.10 g/cm3 in the presence and absence of 0.05% SDS. Gel electrophoresis of the agent in the presence of 0.1% SDS resulted in inactivation of > 95% of the agent loaded onto the gel. Free-flow electrophoresis showed that > 99% of the agent in fraction P5 migrated toward the anode, but not as a discrete species. Sedimentation analysis of the agent in fraction P5 in the presence of 1% lysolecithin showed that the agent has a sedimentation coefficient of < 300S but > 30S. Heating P5 preparations caused the agent to associate with cellular elements and form aggregates with sedimentation coefficients > 10,000S. Removal by differential centrifugation of the large forms of the agent produced upon heating permitted characterization of a discrete subpopulation of scrapie agent particles. Rate-zonal sucrose gradient studies showed that > 95% of the infectivity in this subpopulation sedimented as uniform particles with a sedimentation coefficient of 240S.
MH Animal; Centrifugation, Density Gradient; DNA, Viral/analysis; Detergents/pharmacology; Electrophoresis, Polyacrylamide Gel; Female; Heat; Mice; Prions/*analysis/drug effects; RNA, Viral/analysis; Support, U.S. Gov't, Non-P.H.S.; Support, U.S. Gov't, P.H.S.; Viral Proteins/analysis
AD Stanley B. Prusiner, Department of Neurology and Biochemistry and Biophysics, University of California, San Francisco 94143, California; William J. Hadlow, Richard E. Race, Carl M. Eklund, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840, USA
SP englisch
PO England