Journal article

Engineering a structure switching mechanism into a steroid-binding aptamer and hydrodynamic analysis of the ligand binding mechanism.

Oren Reinstein, Miguel AD Neves, Makbul Saad, Sherry N Boodram, Stephanie Lombardo, Simone A Beckham, Jason Brouwer, Gerald F Audette, Patrick Groves, Matthew CJ Wilce, Philip E Johnson

Biochemistry | Published : 2011

Abstract

The steroid binding mechanism of a DNA aptamer was studied using isothermal titration calorimetry (ITC), NMR spectroscopy, quasi-elastic light scattering (QELS), and small-angle X-ray spectroscopy (SAXS). Binding affinity determination of a series of steroid-binding aptamers derived from a parent cocaine-binding aptamer demonstrates that substituting a GA base pair with a GC base pair governs the switch in binding specificity from cocaine to the steroid deoxycholic acid (DCA). Binding of DCA to all aptamers is an enthalpically driven process with an unfavorable binding entropy. We engineered into the steroid-binding aptamer a ligand-induced folding mechanism by shortening the terminal stem b..

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University of Melbourne Researchers