Chung Chan Ho, Kim Joong Hyun, Chung Bong Hyun
BioNano Health Guard Research Center, Korea Research Institute of Bioscience and Biotechnology , 125 Gwahak-ro, Yuseong-gu, Daejeon 305-806, South Korea.
Anal Chem. 2014 Dec 2;86(23):11586-91. doi: 10.1021/ac503577t. Epub 2014 Nov 19.
In this paper, we report for the first time that graphene oxide (GO) can interact with mutagenic DNA but not intact DNA. After UV-irradiated fluorophore-linked DNA containing thymine repeats was mixed with GO, a decrease in fluorescence was observed in a time-dependent manner. In contrast, no fluorescence change was observed with intact DNA, indicating that UV irradiation of DNA resulted in the formation of mutagenic bases. Because GO is known to act as a fluorescence quencher, the decreased fluorescence implies adsorption of the UV-irradiated DNA onto GO. It appears that the decreased fluorescence might result from the greater accessibility of hydrophobic methyl groups and phenyl rings of thymine dimers to GO and from deformed DNA structures with less effective charge shielding under salt-containing conditions. Using this affinity of GO for mutagenic DNA, we could detect UV-irradiated DNA at concentrations as low as 100 pM. We were also able to analyze the ability of phototoxic drugs to catalyze the formation of mutagens under UV irradiation with GO. Because our method is highly sensitive and feasible and does not require the pretreatment of DNA, we propose that it could accelerate the screening of potential phototoxic drug candidates that would be able to sensitize mutagenic dsDNA.
在本文中,我们首次报道氧化石墨烯(GO)可与诱变DNA相互作用,但不与完整DNA相互作用。将紫外线照射过的含胸腺嘧啶重复序列的荧光团连接DNA与GO混合后,观察到荧光随时间呈下降趋势。相比之下,完整DNA未观察到荧光变化,这表明DNA的紫外线照射导致了诱变碱基的形成。由于已知GO可作为荧光猝灭剂,荧光降低意味着紫外线照射过的DNA吸附到了GO上。荧光降低似乎可能是由于胸腺嘧啶二聚体的疏水甲基和苯环比完整DNA更易接近GO,以及在含盐条件下DNA结构变形导致电荷屏蔽效果降低所致。利用GO对诱变DNA的这种亲和力,我们能够检测低至100 pM浓度的紫外线照射过的DNA。我们还能够分析光毒性药物在紫外线照射下与GO催化诱变剂形成的能力。由于我们的方法高度灵敏且可行,且无需对DNA进行预处理,我们认为它可以加速对能够使诱变双链DNA敏感的潜在光毒性药物候选物的筛选。