Quantitative detection ofAg+ and cysteine using G-quadruplex–hemin DNAzymes

Abstract

A highly sensitive and selective Ag⁺ detection method was developed based on the Ag⁺-mediated formation of G-quadruplex–hemin DNAzymes. In this method, two unlabelled oligonucleotides with different lengths are used. In the absence of Ag⁺, the two oligonucleotides hybridize to each other to form an intermolecular duplex. The addition of Ag⁺ can disrupt the intermolecular duplex and promote a part of the sequence of the longer oligonucleotide to fold into an intramolecular duplex, in which cytosine–cytosine (C–C) mismatches are stabilized by C–Ag⁺–C base pairs. As a result, the G-rich sequence of the same oligonucleotide can fold into a G-quadruplex, which is able to bind hemin to form a catalytically active G-quadruplex–hemin DNAzyme. This can be reflected by an absorbance increase when monitored in the H₂O₂–ABTS (2,2′-azinobis(3-ethylbenzothiozoline)-6-sulfonic acid) reaction system by using UV-vis absorption spectroscopy. This ‘turn-on’ process allows the detection of aqueous Ag⁺ at concentrations as low as 20 nM using a simple colorimetric technique. Considering that Cysteine (Cys) is a strong binder of Ag⁺, the presence of Cys may disrupt the C–Ag⁺–C base pairs in the intramolecular duplex, resulting in the reformation of the intermolecular duplex and the decrease of the catalytic activity of the sensing system. Therefore, the Ag⁺-sensing system can be further developed as a Cys-sensing system. This method allows the detection of Cys with a detection limit of 25 nM. With the development of the studies on DNA–metal base pairs, this Ag⁺-sensing method can be easily extended to the analysis of other metal ions.