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[染料木黄酮的铬(III)配合物与DNA的相互作用研究]

[Research on the interaction of Cr(III) complex of genistein with DNA].

作者信息

Yu Yan-Ying, Li Hua, Hu Xin, Cao Shu-Wen

机构信息

Department of Chemistry, Nanchang University, Nanchang 330031, China.

出版信息

Guang Pu Xue Yu Guang Pu Fen Xi. 2008 Jul;28(7):1587-91.

Abstract

The interaction of the Cr(III) complex of genistein (GEN-Cr) with calf thymus DNA (ctDNA) in Tris (pH 7.2) buffer was investigated using UV spectra, DNA melting, fluorescence spectra and viscosity. From the absorption titration experiment, no obvious red shift was found, but the notable hypochromicities were observed. When C(DNA)/C(GEN-Cr) = 3, the pi-pi* transitions of the complex at 272 nm showed a decrease in intensity of 29.1%, which indicated that there was remarkable intercalation between complex and DNA base pairs, involving a strong pi-stacking interacting between them. The binding constant for the complex was K = 1.9 x 10(5) mol x L(-1). From the melting curves of ctDNA in the absence and presence of the complex, the melting temperature of ctDNA was found to increase by 5.5 degrees C from 74 to 79.5 degrees C, owing to the increased stability of the helix in the presence of the complex that was intercalated into the double helix. The complex could emit weak luminescence in Tris buffer. The emission intensity of the complex at 340 nm increased steadily with the addition of ctDNA. The result suggested that the complex got into a hydrophobic environment inside the DNA and avoided the effect of solvent water molecules. The strong interaction of the complex and ctDNA also resulted in greatly enhanced intensity of the resonance light scattering spectra. The emission intensity of DNA-EB system at 600 nm decreased remarkably with increasing the complex concentration, which indicated that the complex could be intercalated into DNA and replace EB from the DNA-EB system. According to the classical Stern-Volmer equation, the quenching plots at 25 and 37 degrees C both appeared approximately linear. These results showed that there was one predominant quenching style in this process. Viscosity experiments were carried out by an Ubbelodhe viscometer at 20.0 (+/- 0.1) degrees C. The relative viscosity of ctDNA increased steadily with the increased in the complex. The result clearly showed that the complex could be intercalated between DNA base pairs, causing an extension of the helix, and thus increased the viscosity of DNA. The results above indicated that there is a relatively strong interaction between the GEN-Cr complex and ctDNA, and the complex could bind ctDNA mainly by intercalation. The research suggested that the GEN-Cr complex may be a promising candidate for anticancer, which deserves further research.

摘要

利用紫外光谱、DNA熔解、荧光光谱和粘度法研究了染料木黄酮的Cr(III)配合物(GEN-Cr)与小牛胸腺DNA(ctDNA)在Tris(pH 7.2)缓冲液中的相互作用。从吸收滴定实验可知,未发现明显的红移,但观察到显著的减色效应。当C(DNA)/C(GEN-Cr)=3时,配合物在272 nm处的π-π*跃迁强度降低了29.1%,这表明配合物与DNA碱基对之间存在显著的嵌入作用,涉及它们之间强烈的π-堆积相互作用。配合物的结合常数为K = 1.9×10⁵ mol·L⁻¹。从有无配合物存在时ctDNA的熔解曲线可知,由于嵌入到双螺旋中的配合物使螺旋稳定性增加,ctDNA的熔解温度从74℃升高到79.5℃,升高了5.5℃。配合物在Tris缓冲液中能发出微弱的荧光。随着ctDNA的加入,配合物在340 nm处的发射强度稳步增加。结果表明配合物进入了DNA内部的疏水环境,避免了溶剂水分子的影响。配合物与ctDNA的强相互作用还导致共振光散射光谱强度大大增强。随着配合物浓度的增加,DNA-EB体系在600 nm处的发射强度显著降低,这表明配合物可嵌入DNA并从DNA-EB体系中取代EB。根据经典的Stern-Volmer方程,25℃和37℃下的猝灭曲线均近似呈线性。这些结果表明在此过程中存在一种主要的猝灭方式。在20.0(±0.1)℃下用乌氏粘度计进行粘度实验。ctDNA的相对粘度随着配合物的增加而稳步增加。结果清楚地表明配合物可嵌入DNA碱基对之间,导致螺旋伸长,从而增加了DNA的粘度。上述结果表明GEN-Cr配合物与ctDNA之间存在较强的相互作用,且配合物主要通过嵌入作用与ctDNA结合。该研究表明GEN-Cr配合物可能是一种有前景的抗癌候选物,值得进一步研究。

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