In vitro selection or SELEX has allowed for the identification of functional nucleic acids (FNAs) that can potentially mimic and replace protein enzymes. These FNAs likely interact with cofactors, just like enzymes bind cofactors in their active sites. Investigating how FNA binding affects cofactor properties is important for understanding how an active site is formed and for developing useful enzyme mimics. Oxidoreductase enzymes contain cofactors in their active sites that allow the enzymes to do redox chemistry. In certain applications, these redox cofactors act as electron-transfer shuttles that transport electrons between the enzymes’ active sites and electrode surfaces. Three redox cofactors commonly found in oxidoreductases are flavin adenine dinucleotide, nicotinamide adenine dinucleotide (NAD+), and pyrroloquinoline quinone (PQQ). We are interested in investigating how DNA aptamers that bind these cofactors influence the cofactors’ redox abilities and if these aptamer-cofactor complexes could serve as redox catalysts. We employed cyclic voltammetry and amperometry to study the electrochemical properties of NAD+ and PQQ when bound to DNA aptamers. Our results suggest that the aptamers provide a stable environment for the cofactor to participate in redox reactions, although enhanced redox activity was not observed. This work provides a foundation for the development of new FNAs capable of redox activity.
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The authors thank Dr. Tomasz Heyduk for assistance with the fluorescence anisotropy studies. The authors also thank members of the Baum Lab for technical assistance, especially Praveen Bagavandoss and Lucy Freitag who worked in the lab as part of the Students and Teachers as Research Scientists (STARS) program administered by the University of Missouri – St. Louis.
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The authors declare that they have no conflict of interest.
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Emahi, I., Gruenke, P.R. & Baum, D.A. Effect of Aptamer Binding on the Electron-Transfer Properties of Redox Cofactors. J Mol Evol 81, 186–193 (2015). https://doi.org/10.1007/s00239-015-9707-7