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DNA与镍的水溶性复合物的相互作用及DNA交联的形成。

Interaction of DNA with water soluble complex of Nickle and formation of DNA cross-links.

作者信息

Nejat Dehkordi Maryam, Akerman Bjorn

机构信息

Department of Basic Science, Islamic Azad University, Shahrekord Branch, Shahrekord, Iran; Department of Chemical and Biological Engineering, Chalmers University of Technology, SE-412 96, Gothenburg, Sweden.

Department of Chemical and Biological Engineering, Chalmers University of Technology, SE-412 96, Gothenburg, Sweden.

出版信息

Chem Biol Interact. 2018 Feb 25;282:55-62. doi: 10.1016/j.cbi.2018.01.007. Epub 2018 Jan 8.

Abstract

Interaction of double stranded DNA with bulky and hydrophobic Salen type Schiff base complex: [N, N' Bis [3- tert-butyl-5-[triphenyl-phosphonium - methyl] - salicylidene] 1,2 ethylene-diamine nickel(III) acetate (refer to Ni Salen complex) was extensively investigated using the spectroscopic techniques and gel electrophoresis. Absorption titration experiment showed the hypochromic effect and the significant red shift of the complex absorption. In competition experiments with ethidium bromide (EB), Ni Salen complex exhibited non-competitive binding at high concentrations. UV-vis absorption and fluorescence emission data agreed on a binding constant of (1.64 ± 0.01) μM, thereby showing the strong interaction of the complex with DNA; also, a binding site size of 2.33 ± 0.01 base pairs per complex was achieved. Thermal denaturation experiment showed that T of calf thymus-DNA was increased by approximately 10 °C at a molar ratio of the dye/base of 0.2. The CD spectra of DNA exhibited an increase in both positive and negative peaks without any shift in the position of bands upon addition of the complex. The amplitude of the LD spectra of DNA was decreased in the presence of the complex. Reduced linear dichroism (LD) revealed that the transition moment of complex was parallel to the DNA helix axis. Gel electrophoresis experiments confirmed that Ni Salen complex had no nuclease/DNA cleaving activity; also, DNA-DNA cross links were formed at high concentrations of complex, leading to the aggregation of DNA.

摘要

双链DNA与体积较大且具疏水性的Salen型席夫碱配合物的相互作用:[N, N'-双[3-叔丁基-5-[三苯基-鏻-甲基]-亚水杨基]1,2-乙二胺镍(III)乙酸盐(简称Ni Salen配合物),运用光谱技术和凝胶电泳进行了广泛研究。吸收滴定实验表明存在减色效应以及配合物吸收峰的显著红移。在与溴化乙锭(EB)的竞争实验中,Ni Salen配合物在高浓度时表现出非竞争性结合。紫外可见吸收和荧光发射数据得出结合常数为(1.64 ± 0.01) μM,从而表明该配合物与DNA有强烈相互作用;此外,每个配合物的结合位点大小为2.33 ± 0.01个碱基对。热变性实验表明,当染料/碱基的摩尔比为0.2时,小牛胸腺DNA的熔解温度升高了约10°C。加入配合物后,DNA的圆二色光谱在正负峰均有所增加,且谱带位置无任何偏移。在配合物存在的情况下,DNA的线性二色性(LD)光谱幅度降低。降低的线性二色性显示配合物的跃迁矩与DNA螺旋轴平行。凝胶电泳实验证实Ni Salen配合物没有核酸酶/DNA切割活性;此外,在高浓度配合物时会形成DNA-DNA交联,导致DNA聚集。

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