Li Junfen, Shuang Shaomin, Dong Chuan
Department of Chemistry, Center of Environmental Science and Engineering Research, Shanxi University, Taiyuan, China.
Talanta. 2009 Jan 15;77(3):1043-9. doi: 10.1016/j.talanta.2008.08.012. Epub 2008 Aug 27.
The spectroscopic characterizations of solid substrate room temperature phosphorescence (SS-RTP) of palmatine (Pal) have been studied. Strong RTP signal at 615nm can be induced on filter paper in the presence of TIAc. The interaction between calf thymus DNA (ctDNA) and Pal has been investigated at pH 6.90 using fluorescence, UV-vis, SS-RTP and cyclic voltammogram spectroscopy. Strong binding affinity of Pal with DNA is revealed from the absorption and fluorescence studies in the liquid state. With the addition of ctDNA, the fluorescence intensity of Pal is enhanced greatly and UV-vis spectra show no apparent hypochromicity and red shift, which indicates that Pal intercalates into ctDNA bases. However, this conclusion could not explain the phenomena from fluorescence polarization and denatured DNA measurements, which indicate that groove binding is at least the main binding mode. Binding constant and binding site size have been calculated to be 2.57x10(4)L/mol and 0.16 based on Scatchard plot from fluorescence titration data. Groove binding has also been supported by phosphorescence lifetime and anion quenching experiments. Above studies demonstrate that there should exist intercalative binding and groove binding in the interaction of Pal and DNA. Furthermore, cyclic voltammogram study suggests that electrostatic binding exists at the same time exactly. Taken together, the binding model obtained in this study is mixed-mode.
研究了巴马汀(Pal)的固体基质室温磷光(SS-RTP)光谱特性。在三碘乙酸(TIAc)存在下,可在滤纸上诱导出615nm处的强RTP信号。在pH 6.90条件下,利用荧光、紫外可见、SS-RTP和循环伏安光谱研究了小牛胸腺DNA(ctDNA)与Pal之间的相互作用。从液态下的吸收和荧光研究中揭示了Pal与DNA具有很强的结合亲和力。加入ctDNA后,Pal的荧光强度大大增强,紫外可见光谱未显示明显的减色和红移,这表明Pal插入到ctDNA碱基中。然而,这一结论无法解释荧光偏振和变性DNA测量的现象,这些现象表明沟槽结合至少是主要的结合模式。根据荧光滴定数据的Scatchard图计算出结合常数和结合位点大小分别为2.57×10⁴L/mol和0.16。磷光寿命和阴离子猝灭实验也支持沟槽结合。上述研究表明,Pal与DNA相互作用中应存在插入结合和沟槽结合。此外,循环伏安研究表明同时确实存在静电结合。综上所述,本研究获得的结合模型为混合模式。