Molecular mass, biochemical composition, and physicochemical behavior of the infectious form of the scrapie precursor protein monomer.
A highly purified fraction obtained from scrapie (263-K strain)-infected hamsters' brains by an alternative procedure without proteinase K treatment contained a protease-resistant form of the scrapie precursor protein (PrPSc) and infectivity of 9.9 +/- 0.7 log LD50/ml. Polyclonal antibodies produced against hamster scrapie amyloid protein (PrP27-30) and used in a neutralization test diminished infectivity of the PrPSc preparations by 1.6 log after intracerebral inoculation and by 1 log after intraperitoneal inoculation. PrPSc was subjected to size-exclusion HPLC; greater than or equal to 60% of the eluted infectious units were recovered from the peak with an apparent mass of 30.4 +/- 0.6 kDa. Characterization by UV absorption spectra, SDS/PAGE, immunoblots, N-terminal amino acid sequence, and neutral sugar and amino sugar analyses demonstrated homogeneity of the infectious units. The neutral sugar and amino sugar compositional analyses revealed high mannose, glucosamine, fucose, and sialic acid content. This demonstrated an extensive posttranslational modification by the complex type of N-linked glycosylation and glycane core of C-terminal glycolipid of PrPSc. The results correspond to the predicted size, composition, and sequence of PrPSc and indicate that this protein may be the only component of scrapie infectious unit or the infectious form of scrapie precursor.
Safar
J
J
Laboratory of Central Nervous System Studies, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892.
Wang
W
W
Padgett
M P
MP
Ceroni
M
M
Piccardo
P
P
Zopf
D
D
Gajdusek
D C
DC
Gibbs
C J
CJ
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001921:Brain / Q000382:microbiology; D002241:Carbohydrates / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D008647:Mesocricetus; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009500:Neutralization Tests; D010743:Phospholipids; D018620:PrPSc Proteins; D011328:Prions / Q000254:growth & development*; D011498:Protein Precursors / Q000302:isolation & purification*; D014764:Viral Proteins / Q000302:isolation & purification*
D000596:Amino Acids; D002241:Carbohydrates; D010743:Phospholipids; D018620:PrPSc Proteins; D011328:Prions; D011498:Protein Precursors; D014764:Viral Proteins