Title
Rational design of a structure-switching DNA aptamer for potassium ions
Author
Andrew T. Catherine, Shephanie N. Shishido, Gregga A. Robbins-Welty, Amy Diegelman-Parente
Year
2014
Journal
FEBS Open Bio
Abstract
Structure-switching molecules provide a unique means for analyte detection, generating a response to analyte concentration through a binding-specific conformational change between non-binding and binding-competent states. While most ligand-binding molecules are not structure switching by default, many can be engineered to be so through the introduction of an alternative non-binding (and thus non-signalling) conformation. This population-shift mechanism is particularly effective with oligonucleotides and has led to the creation of structure-switching aptamers for many target ligands. Here, we report the rational design of structure-switching DNA aptamers, based on the thrombin binding aptamer (TBA), that bind potassium with affinities that bridge the gap between previously reported weak-binding and strong-binding aptamers. We also demonstrate a correlation between the free energy of the experimentally determined binding affinity for potassium and the computationally estimated free energy of the alternative (non-binding) structure.
Instrument
J-810
Keywords
Circular dichroism, DNA structure, Ligand binding, Protein folding, Biochemistry