AI Article Synopsis
- Selective binding of ligands at the 5' and 3' ends of G-quadruplexes (G4s) can influence their physiological roles, but existing methods to assess this binding are not efficient or affordable.
- A new protocol called G4-AFQ is introduced to measure ligand selectivity on G4s by attaching a fluorophore, AMCA, to either end and observing which end's fluorescence is quenched.
- G4-AFQ offers several advantages over traditional techniques like nuclear magnetic resonance (NMR), including increased convenience, sensitivity, lower cost, and broader applicability for evaluating G4 ligands.
Article Abstract
The selective binding of ligand molecules towards the 5' and 3' ends of G-quadruplex (G4) may differentially affect the physiological function of G4s. However, there is still a lack of sensitive and low-cost approaches to accurately measure the binding preference of ligands on G4s, although multiple ways have been developed to evaluate the interaction between ligands and G4s. Here, we propose a new protocol named G4-AFQ to test the selectivity of ligands towards the two terminal G-tetrads of G4s. In this protocol, the fluorophore AMCA is respectively modified at the 5' or 3' end of G4, and which end of AMCA fluorescence is quenched means that the ligand binds to the G-tetrad at that end. Through G4-AFQ, the affinity constant of ligands towards the binding site can also be obtained. Compared with the commonly used nuclear magnetic resonance (NMR) method, G4-AFQ is more convenient, sensitive, cost-effective, and suitable for the measurement of the vast majority of G4 ligands, with a great potential for widespread application.
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| http://dx.doi.org/10.1016/j.talanta.2024.126732 | DOI Listing |
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