Protease-resistant human prion protein and ferritin are cotransported across Caco-2 epithelial cells: implications for species barrier in prion uptake from the intestine.
Foodborne transmission of bovine spongiform encephalopathy (BSE) to humans as variant Creutzfeldt-Jakob disease (CJD) has affected over 100 individuals, and probably millions of others have been exposed to BSE-contaminated food substances. Despite these obvious public health concerns, surprisingly little is known about the mechanism by which PrP-scrapie (PrP(Sc)), the most reliable surrogate marker of infection in BSE-contaminated food, crosses the human intestinal epithelial cell barrier. Here we show that digestive enzyme (DE) treatment of sporadic CJD brain homogenate generates a C-terminal fragment similar to the proteinase K-resistant PrP(Sc) core of 27-30 kDa implicated in prion disease transmission and pathogenesis. Notably, DE treatment results in a PrP(Sc)-protein complex that is avidly transcytosed in vesicular structures across an in vitro model of the human intestinal epithelial cell barrier, regardless of the amount of endogenous PrP(C) expression. Unexpectedly, PrP(Sc) is cotransported with ferritin, a prominent component of the DE-treated PrP(Sc)-protein complex. The transport of PrP(Sc)-ferritin is sensitive to low temperature, brefeldin-A, and nocodazole treatment and is inhibited by excess free ferritin, implicating a receptor- or transporter-mediated pathway. Because ferritin shares considerable homology across species, these data suggest that PrP(Sc)-associated proteins, in particular ferritin, may facilitate PrP(Sc) uptake in the intestine from distant species, leading to a carrier state in humans.
Mishra
Ravi Shankar
RS
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Basu
Subhabrata
S
Gu
Yaping
Y
Luo
Xiu
X
Zou
Wen-Quan
WQ
Mishra
Richa
R
Li
Ruliang
R
Chen
Shu G
SG
Gambetti
Pierluigi
P
Fujioka
Hisashi
H
Singh
Neena
N
D000368:Aged; D001921:Brain / Q000378:metabolism; D018938:Caco-2 Cells; D007562:Creutzfeldt-Jakob Syndrome / Q000378:metabolism; D005260:Female; D005293:Ferritins / Q000378:metabolism*; D006801:Humans; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D046912:Multiprotein Complexes / Q000378:metabolism; D010434:Pepsin A / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism; D018621:PrPC Proteins / Q000378:metabolism; D018620:PrPSc Proteins / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D013045:Species Specificity; D019108:Tight Junctions / Q000378:metabolism
D046912:Multiprotein Complexes; D010446:Peptide Fragments; D018621:PrPC Proteins; D018620:PrPSc Proteins; D005293:Ferritins; D010447:Peptide Hydrolases; D010434:Pepsin A