Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins.
Scrapie and Creutzfeldt-Jakob disease are transmissible, degenerative neurological diseases caused by prions. Considerable evidence argues that prions contain protease-resistant sialoglycoproteins, designated PrPSc, encoded by a cellular gene. The prion protein (PrP) gene also encodes a normal cellular protein designated PrPC. We established clonal cell lines which support the replication of mouse scrapie or Creutzfeldt-Jakob disease prions. Mouse neuroblastoma N2a cells were exposed to mouse scrapie prions and subsequently cloned. After limited proteinase K digestion, three PrP-immunoreactive proteins with apparent molecular masses ranging between 20 and 30 kilodaltons were detected in extracts of scrapie-infected N2a cells by Western (immuno-) blotting. The authenticity of these PrPSc molecules was established by using monospecific antiserum raised against a synthetic peptide corresponding to a portion of the prion protein. Those clones synthesizing PrPSc molecules possessed scrapie prion infectivity as measured by bioassay; clones without PrPSc failed to demonstrate infectivity. Detection of PrPSc molecules in scrapie-infected N2a cells supports the contention that PrPSc is a component of the infectious scrapie particle and opens new approaches to the study of prion diseases.
Butler
D A
DA
Department of Neurology, University of California, San Francisco 94143-0518.
Scott
M R
MR
Bockman
J M
JM
Borchelt
D R
DR
Taraboulos
A
A
Hsiao
K K
KK
Kingsbury
D T
DT
Prusiner
S B
SB
D000818:Animals; D000911:Antibodies, Monoclonal; D001681:Biological Assay; D002999:Clone Cells; D007562:Creutzfeldt-Jakob Syndrome / Q000235:genetics; D006224:Cricetinae; D019286:Endopeptidase K; D007163:Immunosorbent Techniques; D009447:Neuroblastoma / Q000378:metabolism / Q000382:microbiology*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011328:Prions; D012608:Scrapie / Q000235:genetics*; D012697:Serine Endopeptidases / Q000378:metabolism; D012795:Sialoglycoproteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D011328:Prions; D012795:Sialoglycoproteins; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D019286:Endopeptidase K