Open AccessPurification and properties of the cellular and scrapie hamster prion proteins
Author(s) -
TURK Eric,
TEPLOW David. B.,
HOOD Leroy E.,
PRUSINER Stanley B.
Publication year - 1988
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1988.tb14246.x
Subject(s) - scrapie , hamster , prion protein , prion proteins , microbiology and biotechnology , fungal prion , virology , chemistry , biology , medicine , pathology , disease
During scrapie infection an abnormal isoform of the prion protein (PrP), designated PrP sc , accumulates and is found to copurify with infectivity; to date, no nucleic acid has been found which is scrapie‐sepecific. Both uninfected and scrapie‐infected cells synthesize a Prp isoform, denoted PrP c , which exhibits physical properties that differntiate it from PrP sc was purified by immunoaffinity chromatography using a PrP‐ sepecific monoclonal antibody cross‐linked to protein‐A‐Avidgel. PrP sc was purified by detergent extraction, poly(ethylene glycol) precipation and repeated differentila centrifugation of PrP sc polymers. Both PrP isoforms were found to have the same N‐terminal amino acid sequence which begins at a predicted signal peptide cleavage site. The first 8 residues of PrP c found to be KKXPKPGG and the first 29 residues of PrP c were found to be KKXPKPGGWNTGGSXYPGQGSPGGNRYPP. Arg residue 3 and 15 in PrP sc and 3 PrP c appear to be modified since no contain an intramolecular disulfide bond, liking Cys 179 and 214, which creates a loop of 36 amino acid containing the two N‐ linked glycosylation sites. Developement of purification of how PrP sc is synthesized either form PrP c or a precursor.