NR AYII
AU Sekiya,S.; Noda,K.; Nishikawa,F.; Yokoyama,T.; Kumar,P.K.; Nishikawa,S.
TI Characterization and application of a novel RNA aptamer against the mouse prion protein
QU Journal of Biochemistry 2006 Mar; 139(3): 383-90
PT journal article; research support, non-u.s. gov't
AB In order to isolate RNA aptamers against the mouse prion protein (mPrP), we carried out in vitro selection from RNA pools containing a 30-nucleotide randomized region. Aptamer 60-3 was found to have a high affinity for mPrP (K(d) = 5.6 +/- 1.5 nM), and 2'-fluoro-pyrimidine modifications for RNase resistance did not abolish its binding activity (K(d) = 22 +/- 4 nM). Following 5' biotinylation, aptamer 60-3 specifically detected PrP in mouse brain homogenate in a Northwestern blotting assay. To determine the mPrP-aptamer binding region, we performed protein-deletion-mutant analysis and competition-binding analysis using heparin. The results showed that aptamer 60-3 appears to have binding sites located between amino acids 23-108.
MH Amino Acid Sequence; Animals; Aptamers, Nucleotide/chemistry/*metabolism; Mice; Molecular Sequence Data; Peptide Fragments/metabolism; Prions/*metabolism; Protein Binding/physiology; RNA/chemistry/*metabolism
AD Cooperative Graduate School, University of Tsukuba, Tennodai, Tsukuba, Ibaraki.
SP englisch
PO Japan