Methylene blue as a G-quadruplex binding probe for label-free homogeneous electrochemical biosensing.
Methylene blue as a G-quadruplex binding probe for label-free homogeneous electrochemical biosensing.
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DOI：
10.1021/ac502540m
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发表时间：
2014-09
影响因子：
7.4
通讯作者：
Fang-ting Zhang;Ji Nie;De-Wen Zhang;Ji-Tao Chen;Yinglin Zhou;Xinxiang Zhang
中科院分区：
文献类型：
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作者：
Fang-ting Zhang;Ji Nie;De-Wen Zhang;Ji-Tao Chen;Yinglin Zhou;Xinxiang Zhang
Herein, G-quadruplex sequence was found to significantly decrease the diffusion current of methylene blue (MB) in homogeneous solution for the first time. Electrochemical methods combined with circular dichroism spectroscopy and UV-vis spectroscopy were utilized to systematically explore the interaction between MB and an artificial G-quadruplex sequence, EAD2. The interaction of MB and EAD2 (the binding constant, K ≈ 1.3 × 10(6) M(-1)) was stronger than that of MB and double-stranded DNA (dsDNA) (K ≈ 2.2 × 10(5) M(-1)), and the binding stoichiometry (n) of EAD2/MB complex was calculated to be 1.0 according to the electrochemical titration curve combined with Scatchard analysis. MB was proved to stabilize the G-quadruplex structure of EAD2 and showed a competitive binding to G-quadruplex in the presence of hemin. EAD2 might mainly interact with MB, a positive ligand of G-quadruplex, through the end-stacking with π-system of the guanine quartet, which was quite different from the binding mechanism of dsDNA with MB by intercalation. A novel signal read-out mode based on the strong affinity between G-quadruplex and MB coupling with aptamer/G-quadruplex hairpin structure was successfully implemented in cocaine detection with high specificity. G-quadruplex/MB complex will function as a promising electrochemical indicator for constructing homogeneous label-free electrochemical biosensors, especially in the field of simple, rapid, and noninvasive biochemical assays.
