NR AVDQ
AU Inoue,Y.; Yoshida,M.
TI In vitro assay for fragmentation of amyloid fibers of yeast prion protein
QU Methods (San Diego, Calif.) 2006 May; 39(1): 56-60
PT journal article
AB In prion propagation, fragmentation of amyloid fibers, as well as conformational conversion of prion protein, is critical: the latter increases the net amount of abnormal prion proteins and the former multiplies number of seeds. We present here a method for in vitro measurement of fragmentation of amyloid fibers of yeast Sup35 prion protein. In this method, amyloid fibers are tethered to the surface of magnetic beads. Fragmentation of the fibers results in release of fiber fragments into the medium, which are then quantified by immunoblotting. This method is versatile for other amyloid fibers.
MH Algorithms; Amyloid/*chemistry; Biotin/chemistry; Blotting, Western; Kinetics; Magnetics; Microspheres; Peptide Fragments/chemistry/metabolism; Prions/*chemistry; Recombinant Proteins/chemistry; Saccharomyces cerevisiae/*chemistry; Saccharomyces cerevisiae Proteins/chemistry; Streptavidin/chemistry
AD Chemical Resources Laboratory, Tokyo Institute of Technology, 4259 Nagatsutacho, Yokohama 226-8503, Japan.
SP englisch
PO USA