NR ABLO
AU Bolton,D.C.; Meyer,R.K.; Prusiner,S.B.
TI Scrapie PrP 27-30 is a sialoglycoprotein
QU Journal of Virology 1985 Feb; 53(2): 596-606
IA http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=3918176
PT journal article
AB The major scrapie prion protein, designated PrP 27-30, exhibited both charge and size heterogeneity after purification from infected hamster brains. Eight or more discrete charge isomers of PrP 27-30 with isoelectric points ranging from approximately pH 4.6 to 7.9 were found by using non-equilibrium pH gradient electrophoresis in the first dimension followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. The charge isomers were detected by silver staining as well as by radioiodination. The procedures used to disaggregate PrP 27-30 before electrophoresis in the first dimension do not appear to be responsible for the charge heterogeneity. However, heating PrP 27-30 to 100 degrees C for 15 min in 0.1 N NaOH or 0.1 N HCl resulted in modification of the protein and alteration of its electrophoretic pattern. A PrP 27-30 fragment (molecular weight, 17,100 to 21,900) obtained by cyanogen bromide cleavage also exhibited charge and size heterogeneity. Periodic acid-Schiff staining of PrP 27-30 electrophoresed into sodium dodecyl sulfate-polyacrylamide gels demonstrated that carbohydrate residues are attached to the protein. Digestion of PrP 27-30 with neuraminidase and endo-beta-N-acetylglucosaminidase H resulted in significant changes in the isoelectric pH of PrP 27-30 isomers, whereas digestion with alkaline phosphatase had no effect. Our results demonstrate that PrP 27-30 is a sialoglycoprotein; this is consistent with several properties of this protein and of the scrapie prion.
MH Alkaline Phosphatase; Centrifugation, Density Gradient; Chemistry; Cyanogen Bromide; Electrophoresis, Polyacrylamide Gel; Glycoside Hydrolases; Hydrogen-Ion Concentration; Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; Molecular Weight; Neuraminidase; Periodic Acid-Schiff Reaction; Prions/*analysis; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.; Sialoglycoproteins/*analysis/isolation & purification; Viral Proteins/*analysis/isolation & purification
SP englisch
PO USA