A Universal Nucleoside with Strong Two-Band Switchable Fluorescence and Sensitivity to the Environment for Investigating DNA Interactions
A Universal Nucleoside with Strong Two-Band Switchable Fluorescence and Sensitivity to the Environment for Investigating DNA Interactions
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DOI：
10.1021/ja3030388
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发表时间：
2012-06-20
影响因子：
15
通讯作者：
Burger, Alain
中科院分区：
文献类型：
--
作者：
Dziuba, Dmytro;Postupalenko, Viktoriia Y.;Burger, Alain
为了开发一种新的研究DNA相互作用的工具，合成了一种含有3-羟色酮(3HC)荧光基团的核苷类似物作为碱基模拟物，并将其掺入到寡核苷酸链中。与现有的荧光核苷类似物相比，这种染料具有特殊的环境敏感性，可以在两个分辨率良好的荧光带之间切换。在标记的DNA中，这种核苷类似物不会改变双链构象，并显示出高的荧光量子产率。这种探针的亮度比荧光核苷标准2-氨基嘌呤高50倍。此外，双发射对环境的极性高度敏感，因此观察到侧翼基座对水的强烈屏蔽作用。有了这种核苷，病毒伴侣蛋白对DNA碱基堆积的影响被选择性地监测。
With the aim of developing a new tool to investigate DNA interactions, a nucleoside analogue incorporating a 3-hydroxychromone (3HC) fluorophore as a nucleobase mimic was synthesized and incorporated into oligonucleotide chains. In comparison with existing fluorescent nucleoside analogues, this dye features exceptional environmental sensitivity switching between two well-resolved fluorescence bands. In labeled DNA, this nucleoside analogue does not alter the duplex conformation and exhibits a high fluorescence quantum yield. This probe is up to 50-fold brighter than 2-aminopurine, the fluorescent nucleoside standard. Moreover, the dual emission is highly sensitive to the polarity of the environment; thus, a strong shielding effect of the flanking bases from water was observed. With this nucleoside, the effect of a viral chaperone protein on DNA base stacking was site-selectively monitored.