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Tertiary RNA Folding-Targeted Drug Screening Strategy Using a Protein Nanopore
- Source :
- Analytical Chemistry. 93:2811-2819
- Publication Year :
- 2021
- Publisher :
- American Chemical Society (ACS), 2021.
-
Abstract
- Bacterial riboswitch RNAs are attractive targets for novel antibiotics against antibiotic-resistant superbacteria. Their binding to cognate metabolites is essential for the regulation of bacterial gene expression. Despite the importance of RNAs as therapeutic targets, the development of RNA-targeted, small-molecule drugs is limited by current biophysical methods. Here, we monitored the specific interaction between the adenine-sensing riboswitch aptamer domain (ARS) and adenine at the single-molecule level using α-hemolysin (αHL) nanopores. During adenine-induced tertiary folding, adenine-bound ARS intermediates exhibited characteristic nanopore events, including a two-level ionic current blockade and a ∼ 5.6-fold longer dwell time than that of free RNA. In a proof-of-concept experiment, tertiary RNA folding-targeted drug screening was performed using a protein nanopore, which resulted in the discovery of three new ARS-targeting hit compounds from a natural compound library. Taken together, these results reveal that αHL nanopores are a valuable platform for ultrasensitive, label-free, and single-molecule-based drug screening against therapeutic RNA targets.
- Subjects :
- Drug
Riboswitch
RNA Folding
Chemistry
media_common.quotation_subject
Aptamer
010401 analytical chemistry
Drug Evaluation, Preclinical
A protein
RNA
Computational biology
010402 general chemistry
01 natural sciences
0104 chemical sciences
Analytical Chemistry
Folding (chemistry)
Hemolysin Proteins
Nanopores
Nanopore
Gene expression
media_common
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 93
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....0edbdc7d27bbb196e60b8754d4cd4ae7