A high-throughput fluorescent indicator displacement assay and principal component/cluster data analysis for determination of ligand-nucleic acid structural selectivity
| Autor: | Robert D. Gray, Rafael del Villar-Guerra, Jonathan B. Chaires, John O. Trent |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0303 health sciences
Thermal shift assay Ligand 010402 general chemistry 01 natural sciences Fluorescence Combinatorial chemistry 0104 chemical sciences 03 medical and health sciences chemistry.chemical_compound chemistry Principal component analysis Nucleic acid Selectivity DNA Binding selectivity 030304 developmental biology |
| DOI: | 10.1101/192849 |
| Popis: | We describe a high-throughput fluorescence indicator displacement assay (HT-FID) to evaluate the affinity and the selectivity of compounds binding to different DNA structures. We validated the assay using a library of 30 well-known nucleic acid binders containing a variety chemical scaffolds. We used a combination of principal component analysis and hierarchical clustering analysis to interpret the results obtained. This analysis classified compounds based on selectivity for AT-rich, GC-rich and G4 structures. We used the FID assay as a secondary screen to test the binding selectivity of an additional 20 compounds selected from the NCI diversity set III library that were identified as G4 binders using a thermal shift assay. The results showed G4 binding selectivity for only a few of the 20 compounds. Overall, we show that this HT-FID assay, coupled with PCA and HCA, provides a useful tool for the discovery of ligands selective for particular nucleic acid structures. |
| Databáze: | OpenAIRE |
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