Fluorescent DNA-Protected Silver Nanoclusters for Ligand-HIV RNA Interaction Assay.
Fluorescent DNA-Protected Silver Nanoclusters for Ligand-HIV RNA Interaction Assay.
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DOI：
10.1021/acs.analchem.5b03166
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发表时间：
2015-10
影响因子：
7.4
通讯作者：
Liang Qi;Yuan Huo;Huan Wang;Jing Zhang;Fuquan Dang;Zhiqi Zhang
中科院分区：
文献类型：
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作者：
Liang Qi;Yuan Huo;Huan Wang;Jing Zhang;Fuquan Dang;Zhiqi Zhang
研究配体与生物大分子的相互作用为创造能够有效调节特定生物过程的新化合物提供了机会。核糖核酸（RNA）分子在调节基因表达方面的作用被发现后，已成为有吸引力的药物靶点，而只有有限数量的研究调查了配体与功能性RNA分子之间的相互作用，特别是基于纳米技术的研究。本研究首次使用dna保护银纳米簇（agnc）来研究配体与rna的相互作用。发现蒽环类抗癌药物米托蒽醌（MTX）可猝灭agnc的荧光。在agnc -MTX混合物中加入人类免疫缺陷病毒反式激活反应区（TAR） RNA或Rev-response element (RRE) RNA后，由于MTX与RNA的相互作用，agnc的荧光恢复。通过理论计算确定MTX与TAR和RRE RNA的结合常数和结合位点数量。荧光偏振和质谱实验进一步证实了MTX-RNA的相互作用。探讨了mtx基荧光猝灭agnc的机理。本研究提供了一种新的配体- rna结合相互作用测定方法。
Studying ligand-biomacromolecule interactions provides opportunities for creating new compounds that can efficiently regulate specific biological processes. Ribonucleic acid (RNA) molecules have become attractive drug targets since the discovery of their roles in modulating gene expression, while only a limited number of studies have investigated interactions between ligands and functional RNA molecules, especially those based on nanotechnology. DNA-protected silver nanoclusters (AgNCs) were used to investigate ligand-RNA interactions for the first time in this study. The anthracycline anticancer drug mitoxantrone (MTX) was found to quench the fluorescence of AgNCs. After adding human immunodeficiency virus trans-activation responsive region (TAR) RNA or Rev-response element (RRE) RNA to AgNCs-MTX mixtures, the fluorescence of the AgNCs recovered due to interactions between MTX with RNAs. The binding constants and number of binding sites of MTX to TAR and RRE RNA were determined through theoretical calculations. MTX-RNA interactions were further confirmed in fluorescence polarization and mass spectrometry experiments. The mechanism of MTX-based fluorescence quenching of the AgNCs was also explored. This study provides a new strategy for ligand-RNA binding interaction assay.