NR APVS
AU Sayer,N.M.; Cubin,M.; Rhie,A.; Bullock,M.; Tahiri-Alaoui,A.; James,W.
TI Structural determinants of conformationally selective, prion-binding aptamers
QU The Journal of Biological Chemistry 2004 Mar 26; 279(13): 13102-9
PT journal article
AB We have recently described the isolation of 2'-fluoropyrimidine-substituted RNA aptamers that bind selectively to disease-associated beta-sheet-rich forms of the prion protein, PrP, from a number of mammalian species. These aptamers inhibit the accumulation of protease-resistant forms of PrP in a prion-seeded, in vitro conversion assay. Here we identify the minimal portions of two of these aptamers that retain binding specificity. We determine their secondary structures by a combination of modeling and solution probing. Finally, we identify an internal site for biotinylation of a minimized, synthetic aptamer and use the resultant reagent in the detection of abnormal forms of PrP in vitro.
MH Animals; Base Sequence; Binding Sites; Biotinylation; Cattle; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Escherichia coli/metabolism; Kinetics; Molecular Sequence Data; Prions/*chemistry; Protein Binding; Protein Conformation; Protein Folding; Protein Structure, Secondary; RNA/chemistry; Support, Non-U.S. Gov't; Transcription, Genetic; Urea/pharmacology
AD Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom.
SP englisch
PO USA