Slavkovic, SladjanaChurcher, Zachary RJohnson, Philip E2023-05-102023-05-102018-09-17Bioorganic & Medicinal Chemistry 26 (2018): 5427–5434https://doi.org/10.1016/j.bmc.2018.09.017http://hdl.handle.net/10315/41135An unusual feature of the cocaine-binding aptamer is that it binds quinine much tighter than the ligand it was selected for, cocaine. Here we expand the repertoire of ligands that this aptamer binds to include the quinine- based antimalarial compounds amodiaquine, mefloquine, chloroquine and primaquine. Using isothermal titra- tion calorimetry (ITC) we show that amodiaquine is bound by the cocaine-binding aptamer with an affinity of (7 ± 4) nM, one of the tightest aptamer-small molecule affinities currently known. Amodiaquine, mefloquine and chloroquine binding are driven by both a favorable entropy and enthalpy of binding, while primaquine, quinine and cocaine binding are enthalpy driven with unfavorable binding entropy. Using nuclear magnetic resonance (NMR) and ITC methods we show that these ligands compete for the same binding sites in the ap- tamer. Our identification of such a tight binding ligand for this aptamer should prove useful in developing new biosensor techniques and applications using the cocaine-binding aptamer as a model system.enAttribution-NonCommercial-NoDerivatives 4.0 InternationalAptamer; Isothermal titration calorimetry; NMR spectroscopy; Biomolecular interactionsArticle