Institute of Analytical Chemistry, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, 81237 Bratislava, Slovakia.
Institute of Organic Chemistry, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, 81237 Bratislava, Slovakia.
Bioelectrochemistry. 2018 Oct;123:182-189. doi: 10.1016/j.bioelechem.2018.05.004. Epub 2018 May 3.
The understanding of DNA-drug interaction mechanism is among the important aspects of biological studies for drug design, discovery and pharmaceutical development processes. Published rather detailed FTIR and UV-visible spectroscopic studies on the interactions of theophylline, theobromine and caffeine with calf thymus DNA have shown effective binding of these methylxanthine derivatives to DNA and RNA involving H-bonds. However, to our knowledge, there is no such investigation using electrochemical approach. As a novelty of the study, in this paper the bioelectrochemical approach has been chosen for the investigation of an interaction of low molecular salmon sperm dsDNA, ssDNA and mononucleotides with theophylline (TP) in aqueous phosphate buffered medium using DNA-based electrochemical biosensors and biosensing in solution phase. Exploitation of the electrochemical approach via changes in square wave voltammetric responses of deoxyguanosine (dGuo) and deoxyadenosine (dAdo) provided a new indication on preferential association of TP with dGuo in the case of double helical dsDNA structure which was not reported previously. Moreover, an attachment of TP molecules outside DNA was found in the presence of high concentration of 3.3 × 10 M TP in solution which diminishes the electron transfer and leads to the difficulties in quantitative evaluation of the TP and dGuo voltammetric responses. The changes in UV-vis and FTIR spectra obtained in the same medium confirmed the association interaction of TP with both nucleobases. Utilizing the model and the published energies of hydrogen bonding stabilization, the formation of a DNA-TP complex was predicted through the intermolecular H-bonds between TP and the NH-CO moiety of guanine and the N-NH moiety of adenine.
DNA-药物相互作用机制的理解是药物设计、发现和药物开发过程中生物研究的重要方面之一。已发表的关于茶碱、可可碱和咖啡因与小牛胸腺 DNA 相互作用的相当详细的傅里叶变换红外（FTIR）和紫外-可见光谱研究表明，这些甲基黄嘌呤衍生物通过氢键有效地与 DNA 和 RNA 结合。然而，据我们所知，目前还没有使用电化学方法进行这样的研究。作为这项研究的新颖之处，本文选择生物电化学方法来研究低分子鲑鱼精子 dsDNA、ssDNA 和单核苷酸与茶碱（TP）在含磷酸盐缓冲介质中的相互作用，使用基于 DNA 的电化学生物传感器和溶液相中的生物传感。脱氧鸟苷（dGuo）和脱氧腺苷（dAdo）的方波伏安响应变化的电化学方法的利用提供了一个新的指示，即在双螺旋 dsDNA 结构中，TP 优先与 dGuo 结合，这在以前的报道中没有提到。此外，在溶液中存在高浓度的 3.3×10M TP 的情况下，发现 TP 分子附着在 DNA 之外，这会阻碍电子转移，导致难以对 TP 和 dGuo 的伏安响应进行定量评估。在相同介质中获得的 UV-vis 和 FTIR 光谱的变化证实了 TP 与两种核碱基的缔合相互作用。利用模型和已发表的氢键稳定能，预测了通过 TP 与鸟嘌呤的 NH-CO 部分和腺嘌呤的 N-NH 部分之间的分子间氢键形成 DNA-TP 复合物。
