RNA aptamers generated against oligomeric Abeta40 recognize common amyloid aptatopes with low specificity but high sensitivity.
Aptamers are useful molecular recognition tools in research, diagnostics, and therapy. Despite promising results in other fields, aptamer use has remained scarce in amyloid research, including Alzheimer's disease (AD). AD is a progressive neurodegenerative disease believed to be caused by neurotoxic amyloid beta-protein (Abeta) oligomers. Abeta oligomers therefore are an attractive target for development of diagnostic and therapeutic reagents. We used covalently-stabilized oligomers of the 40-residue form of Abeta (Abeta40) for aptamer selection. Despite gradually increasing the stringency of selection conditions, the selected aptamers did not recognize Abeta40 oligomers but reacted with fibrils of Abeta40, Abeta42, and several other amyloidogenic proteins. Aptamer reactivity with amyloid fibrils showed some degree of protein-sequence dependency. Significant fibril binding also was found for the naïve library and could not be eliminated by counter-selection using Abeta40 fibrils, suggesting that aptamer binding to amyloid fibrils was RNA-sequence-independent. Aptamer binding depended on fibrillogenesis and showed a lag phase. Interestingly, aptamers detected fibril formation with > or =15-fold higher sensitivity than thioflavin T (ThT), revealing substantial beta-sheet and fibril formation undetected by ThT. The data suggest that under physiologic conditions, aptamers for oligomeric forms of amyloidogenic proteins cannot be selected due to high, non-specific affinity of oligonucleotides for amyloid fibrils. Nevertheless, the high sensitivity, whereby aptamers detect beta-sheet formation, suggests that they can serve as superior amyloid recognition tools.
Rahimi
Farid
F
Department of Neurology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, USA.
Murakami
Kazuma
K
Summers
Jamie L
JL
Chen
Chi-Hong B
CH
Bitan
Gal
G
D000544:Alzheimer Disease / Q000378:metabolism; D000682:Amyloid / Q000737:chemistry*; D016229:Amyloid beta-Peptides / Q000737:chemistry*; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D052160:Benzothiazoles; D004277:DNA, Single-Stranded / Q000737:chemistry; D003720:Densitometry / Q000379:methods; D019281:Dimerization; D006801:Humans; D008024:Ligands; D010446:Peptide Fragments / Q000737:chemistry*; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013844:Thiazoles / Q000737:chemistry
D000682:Amyloid; D016229:Amyloid beta-Peptides; D052157:Aptamers, Nucleotide; D052160:Benzothiazoles; D004277:DNA, Single-Stranded; D008024:Ligands; D010446:Peptide Fragments; D013844:Thiazoles; C078592:amyloid beta-protein (1-40); C009462:thioflavin T