Salt-mediated Two-site Ligand Binding by the Cocaine-binding Aptamer

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2016 Dec 28

PMID 28025391

Citations 14

Authors

Miguel A D Neves

Sladjana Slavkovic

Zachary R Churcher

Philip E Johnson

Affiliations

Soon will be listed here.

Abstract

Multisite ligand binding by proteins is commonly utilized in the regulation of biological systems and exploited in a range of biochemical technologies. Aptamers, although widely utilized in many rationally designed biochemical systems, are rarely capable of multisite ligand binding. The cocaine-binding aptamer is often used for studying and developing sensor and aptamer-based technologies. Here, we use isothermal titration calorimetry (ITC) and NMR spectroscopy to demonstrate that the cocaine-binding aptamer switches from one-site to two-site ligand binding, dependent on NaCl concentration. The high-affinity site functions at all buffer conditions studied, the low-affinity site only at low NaCl concentrations. ITC experiments show the two ligand-binding sites operate independently of one another with different affinities and enthalpies. NMR spectroscopy shows the second binding site is located in stem 2 near the three-way junction. This ability to control ligand binding at the second site by adjusting the concentration of NaCl is rare among aptamers and may prove a useful in biotechnology applications. This work also demonstrates that in vitro selected biomolecules can have functions as complex as those found in nature.

Citing Articles

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Xie Y, Huang D, Xu L, Wan T, Cao Y, Salminen K Mikrochim Acta. 2024; 191(9):510.

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Aptamer-based nanotrains and nanoflowers as quinine delivery systems.

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Aptamers as Theragnostic Tools in Prostate Cancer.

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A proof of concept application of aptachain: ligand-induced self-assembly of a DNA aptamer.

Neves M, Slavkovic S, Reinstein O, Shoara A, Johnson P RSC Adv. 2022; 9(3):1690-1695.

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Unraveling the binding mode of a methamphetamine aptamer: A spectroscopic and calorimetric study.

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