Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S.C. Mullick Road, Kolkata 700 032, West Bengal, India.
J Phys Chem B. 2010 May 6;114(17):5851-61. doi: 10.1021/jp909127a.
The DNA binding property of a Cu(II) complex, viz., Cu(mal)(2)(2).2H(2)O, (mal)(2) = malonic acid, picH = protonated 2-amino-4-picoline, has been investigated in this study. The binding of this complex with plasmid and chromosomal DNA has been characterized by different biophysical techniques. From the absorption and fluorescence spectroscopic studies, it has been observed that the said copper complex binds strongly with pUC19 plasmid and CT DNA with a binding affinity of 2.368 x 10(3) and 4.0 x 10(3) M(-1), respectively, in 10 mM citrate-phosphate buffer, pH 7.4. Spectrofluorimetric studies reveal that the copper complex exhibits partial DNA intercalation as well as partial DNA minor groove binding properties. Consequently, in agarose gel electrophoresis study, it has been observed that the complex alone induces positive supercoiling in plasmid DNA while in the presence of H(2)O(2) it exhibits nuclease activity. The induction of the breakage in DNA backbone depends upon the relative concentrations of H(2)O(2) and copper complex followed by the time of incubation with DNA. Optical DNA melting study, isothermal titration calorimetry, and absorption spectroscopy have been used to characterize the nuclease activity of this complex in the presence of H(2)O(2). Further, (1)H NMR study indicates that Cu(II) in the complex is converted into the Cu(I) state by the reduction of H(2)O(2). Finally, agarose gel electrophoresis study with different radical scavengers concludes that the production of both hydroxyl radicals and reactive oxygen species is responsible for this nuclease activity.
本研究考察了一种 Cu(II) 配合物 [Cu(mal)(2)] (picH) (2)·2H(2)O(mal(2)= 丙二酸,picH = 质子化 2-氨基-4-吡啶)的 DNA 结合性质。通过不同的物理化学技术研究了该配合物与质粒和染色体 DNA 的结合。从吸收和荧光光谱研究中,观察到该铜配合物与 pUC19 质粒和 CT DNA 具有很强的结合作用,在 10 mM 柠檬酸盐-磷酸盐缓冲液(pH 7.4)中,其结合亲和力分别为 2.368×10(3)和 4.0×10(3) M(-1)。荧光光谱研究表明,该铜配合物表现出部分 DNA 嵌入和部分 DNA 小沟结合性质。因此,在琼脂糖凝胶电泳研究中,观察到该配合物单独诱导质粒 DNA 产生正超螺旋,而在 H(2)O(2)存在下,它表现出核酸酶活性。DNA 骨架断裂的诱导取决于 H(2)O(2)和铜配合物的相对浓度,以及与 DNA 孵育的时间。光诱导 DNA 熔解研究、等温滴定量热法和吸收光谱法用于研究 H(2)O(2)存在下该配合物的核酸酶活性。此外,(1)H NMR 研究表明,配合物中的 Cu(II)通过 H(2)O(2)的还原转化为 Cu(I)态。最后,不同自由基清除剂的琼脂糖凝胶电泳研究表明,羟基自由基和活性氧物质的产生是导致这种核酸酶活性的原因。