Abstract |
The pathogenesis of prion diseases is characterized by the accumulation of amyloid-like rods or scrapie-associated fibrils. The major protein component of scrapie-associated fibrils is an abnormally folded isoform of the normal cellular prion protein (PrP(C)) that is resistant to digestion by proteinase K and is referred to as PrP(Sc). Purified human recombinant (hr PrP) was used to characterize the binding of a set of RNAs with affinity to PrP proteins. We report here that hr PrP has two RNA-binding activities at physiological pH. One activity is capable of binding all of the screened RNAs with high affinity, whereas the other activity can bind only to a subset of the RNAs with high affinity in the presence of non-specific competitor RNAs. A novel RNA belonging to the latter class, RQ11+12, bound to hr PrP with high affinity in the presence of vast molar excesses of competing RNAs. Beads impregnated with the RQ11+12 RNA were used to construct a filtration column. The column efficiently bound hr PrP and native PrP(C) from serum and urine. Importantly, the filtration device was also capable of binding proteinase K-treated PrP(Sc) from serum and urine. The level of sensitivity of detection of PrP by standard Western blotting was increased at least 1000-fold by first concentrating PrP from solution with the filtration column.
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Authors | Brian Zeiler, Victor Adler, Valentin Kryukov, Abraham Grossman |
Journal | Biotechnology and applied biochemistry
(Biotechnol Appl Biochem)
Vol. 37
Issue Pt 2
Pg. 173-82
(Apr 2003)
ISSN: 0885-4513 [Print] United States |
PMID | 12630906
(Publication Type: Evaluation Study, Journal Article, Research Support, U.S. Gov't, P.H.S.)
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Chemical References |
- Membranes, Artificial
- PrPC Proteins
- RNA
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Topics |
- Chromatography, Affinity
(methods)
- Membranes, Artificial
- PrPC Proteins
(blood, chemistry, isolation & purification, urine)
- Protein Binding
- RNA
(chemical synthesis, chemistry)
- Ultrafiltration
(methods)
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