NR AMHT
AU Weiss,S.; Proske,D.; Neumann,M.; Groschup,M.H.; Kretzschmar,H.A.; Famulok,M.; Winnacker,E.L.
TI RNA aptamers specifically interact with the prion protein PrP
QU Journal of Virology 1997 Nov; 71(11): 8790-7
PT journal article
AB We have isolated RNA aptamers which are directed against the recombinant Syrian golden hamster prion protein rPrP23-231 (rPrPc) fused to glutathione S-transferase (GST). The aptamers did not recognize the fusion partner GST or the fusion protein GST::rPrP90-231 (rPrP27-30), which lacks 67 amino acids from the PrP N terminus. The aptamer-interacting region of PrPc was mapped to the N-terminal amino acids 23 to 52. Sequence analyses suggest that the RNA aptamers may fold into G-quartet-containing structural elements. Replacement of the G residues in the G quartet scaffold with uridine residues destroyed binding to PrP completely, strongly suggesting that the G quartet motif is essential for PrP recognition. Individual RNA aptamers interact specifically with prion protein in brain homogenates from wild-type mice (C57BL/6), hamsters (Syrian golden), and cattle as shown by supershifts obtained in the presence of anti-PrP antibodies. No interaction was observed with brain homogenates from PrP knockout mice (prn-p(0/0)). Specificity of the aptamer-PrP interaction was further confirmed by binding assays with antisense aptamer RNA or a mutant aptamer in which the guanosine residues in the G tetrad scaffold were replaced by uridine residues. The aptamers did not recognize PrP27-30 in brain homogenates from scrapie-infected mice. RNA aptamers may provide a first milestone in the development of a diagnostic assay for the detection of transmissible spongiform encephalopathies.
MH Animal; Base Sequence; Binding Sites; Cattle; Chimeric Proteins; Glutathione Transferase; Hamsters; Mice; Molecular Sequence Data; Nucleic Acid Conformation; Oligoribonucleotides/*chemistry; PrP 27-30 Protein/*chemistry; RNA-Binding Proteins/*chemistry; Species Specificity; Structure-Activity Relationship; Support, Non-U.S. Gov't
AD Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Munich, Germany. Weiss@lmb.uni-muenchen.de
SP englisch
PO USA