Competitive DNA-Binding Studies between Metal Complexes and GelRed as a New and Safe Fluorescent DNA Dye
Competitive DNA-Binding Studies between Metal Complexes and GelRed as a New and Safe Fluorescent DNA Dye
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DOI：
10.1007/s10895-016-1850-z
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发表时间：
2016-07-01
影响因子：
2.7
通讯作者：
Torkzadeh-Mahani, Masoud
中科院分区：
文献类型：
--
作者：
Anjomshoa, Marzieh;Torkzadeh-Mahani, Masoud
本工作的重点是介绍GelRed（GR）作为一种稳定，灵敏和环境安全的荧光DNA染料，而不是高毒性的溴化乙锭（EB）。金属配合物之间的竞争性DNA结合研究，[Cu（phen-dion）（phen）Cl]Cl（1），[Cu（phen-dione）（bpy）Cl]Cl（2）、[Cu（dppt）（2）（H2O）]PF6（3）、[Ni（dppt）（2）Cl-2]（4）、[Zn（dppt）（2）Cl-2]（5）和K-3[Fe（CN）（6）]（6）（其中phen-dione是1，10-菲咯啉-5，6-二酮，phen是1，10-菲咯啉，bpy是2，2 '-联吡啶，dppt是5，6-二苯基-3-（2-吡啶基）-1，2，4-三嗪），和GelRed在生理条件下通过荧光光谱进行了研究。该方法简单易行，能较好地揭示金属配合物与DNA的结合亲和力和结合方式。该方法是基于由金属络合物从DNA中置换GelRed所产生的荧光的减少。荧光降低%与DNA结合的程度直接相关。结果表明，配合物与DNA的结合能力顺序为3 > 4 > 1 > 2 > 5 > 6.显着淬灭的GR-DNA的发射带与另外的配合物1，3，和4表明，配合物竞争的DNA结合位点与GR和取代GR从GR-DNA，这通常是特征的嵌入相互作用的化合物与DNA。一个小淬灭的发射带的GR-DNA与添加的复合物2观察到，显示复合物较弱的竞争与GR的DNA结合位点比复合物1，3和4。结果表明，配合物5和6不能与GR竞争DNA结合位点，它们与DNA的结合是外部结合（沟结合或静电结合）。
The focus of this work is introduction of GelRed (GR) as a stable, sensitive and environmentally safe fluorescent DNA dye instead of the highly toxic ethidium bromide (EB). Competitive DNA-binding studies between metal complexes, [Cu(phen-dion)(phen)Cl]Cl (1), [Cu(phen-dione)(bpy)Cl]Cl (2), [Cu(dppt)(2)(H2O)]PF6 (3), [Ni(dppt)(2)Cl-2] (4), [Zn(dppt)(2)Cl-2] (5), and K-3[Fe(CN)(6)] (6) (where phen-dione is 1,10-phenanthroline-5,6-dione, phen is 1,10- phenanthroline, bpy is 2,2'-bipyridine, and dppt is 5,6-diphenyl-3-(2-pyridyl)-1,2,4-triazine), and GelRed have been investigated under physiological conditions by fluorescence spectroscopy. This simple method can reveal the binding affinity and mode of metal complexes with DNA. The method is based on the decrease of fluorescence derived from the displacement of GelRed from DNA by metal complexes. The % fluorescence decrease is directly related to the extent of DNA binding. Results indicate the DNA binding affinities of complexes follow the order 3 > 4 > 1 > 2 > 5 > 6. The significant quenching of the emission band of the GR-DNA with the addition of complexes 1, 3, and 4 suggests that complexes compete for DNA-binding sites with GR and displace GR from the GR-DNA, which is usually characteristic of the intercalative interaction of compounds with DNA. A small quenching of the emission band of the GR-DNA with the addition of the complex 2 was observed that show the complex weaker competes for DNA-binding sites with GR than complexes 1, 3, and 4. Results show complexes 5 and 6 cannot compete for DNA-binding sites with GR and their interaction with DNA is external binding (groove or electrostatic bindig).