荧光探针法研究百草枯与DNA的相互作用

以中性红（NR）为分子探针,应用紫外光谱法和荧光光谱法研究了除草剂百草枯（PQ）与小牛胸腺DNA（ctDNA）之间的相互作用。在pH7.3的Tris-HCl（5×10-2mol.L-1）缓冲溶液中,NR分子主要以嵌插方式结合到ctDNA双螺旋结构上,百草枯的加入抑制了NR与ctDNA之间的结合,用Stern-Volmer方程进行数据处理,表明百草枯对ctDNA-NR的荧光猝灭不是单纯的静态或动态猝灭方式,属于混合型的。综合紫外光谱、离子强度等研究证实,在该实验条件下,百草枯与DNA之间存在静电和嵌插两种作用。     

染料木素酯化修饰物与ctDNA的相互作用

在pH7.2的Tris-HCl缓冲溶液中,采用紫外光谱和荧光光谱法研究了2种新型染料木素酯化修饰物,染料木素7-乙酰阿魏酸酯(GenA)和染料木素7,4′-二-乙酰阿魏酸酯(GenDA)与小牛胸腺DNA(ctDNA)的相互作用。随着ctDNA的加入,GenA与GenDA的紫外吸收和荧光光谱的强度均发生不同程度的降低。ctDNA对化合物的荧光猝灭为静态猝灭过程。在293K时化合物与ctDNA的结合常数分别为2.81×106L.mol-1和1.19×104L.mol-1。同时通过I-离子效应、离子强度、DNA熔点、粘度法等研究证实,在该实验条件下,GenA与ctDNA之间具有较强的作用,主要以嵌插方式结合;GenDA主要以沟槽方式与ctDNA作用。     

荧光光谱法研究紫草素与DNA的相互作用

在pH 7.4的Tris-HCl介质中,以盐酸小檗碱(BR)为荧光探针,研究了紫草素(SHI)与鲱鱼精DNA的相互作用。结果表明,SHI与BR竞争结合DNA上的位点;SHI对DNA-BR体系的荧光有猝灭作用,猝灭机制为静态猝灭和动态猝灭共存,SHI与DNA之间为嵌插和静电两种作用方式;热变性实验进一步证明嵌插结合是SHI与DNA的主要结合模式。最后由Scatchard方程求得二者的结合常数为3.48×104 L·mol-1。     

氧氟沙星和左氧氟沙星与DNA的相互作用研究

采用紫外光谱、荧光光谱、荧光偏振以及K₃Fe(CN)₆荧光猝灭实验研究了氧氟沙星(Ofloxacin,OFLX)和左氧氟沙星(Levofloxacin,L-OFLX)与小牛胸腺DNA(ctDNA)的相互作用差异性与作用模式。紫外光谱的结果表明,当向OFLX和L-OFLX溶液中加入ctDNA并且浓度增大时,OFLX和L-OFLX的吸收光谱都呈现略微的减色效应,但吸收峰位置没有发生偏移,L-OFLX的减色效应略强于OFLX的减色效应;从荧光光谱以及OFLX和L-OFLX的Scatchard方程,获得其键合常数分别为1.15×105 L·mol-1,3.75×105 L·mol-1,表明L-OFLX与ctDNA的相互作用要略强于OFLX与ctDNA的相互作用;荧光偏振实验、单双链ctDNA与药物作用实验、K₃Fe(CN)₆荧光猝灭实验都表明OFLX、L-OFLX与ctDNA的作用模式可能是沟槽结合。     

荧光光谱法研究拜复乐与小牛胸腺DNA的作用机理

用荧光光谱法研究了拜复乐(MXFX)与小牛胸腺DNA(ctDNA)之间的相互作用。在pH=7.4的TrisHCl缓冲溶液中,MXFX的荧光激发峰和发射峰分别位于291 nm和462 nm。ctDNA的加入对MXFX的荧光有静态猝灭作用,这种荧光静态猝灭作用是由ctDNA和MXFX结合引起的,作用力为氢键或范德华力,结合常数为1.28×105L/mol(25℃)。采用离子强度的影响、碘离子猝灭及溴乙锭竞争作用实验研究了MXFX与ctDNA间的相互作用,结果表明MXFX与DNA的结合是MXFX嵌入到DNA中相邻2个碱基对之间,属于嵌入结合模式。     

荧光光谱法研究拜复乐与小牛胸腺DNA的作用机理

用荧光光谱法研究了拜复乐（MXFX）与小牛胸腺DNA（ctDNA）之间的相互作用。在pH=7.4的Tris-HCl缓冲溶液中,MXFX的荧光激发峰和发射峰分别位于291 nm和462 nm。ctDNA的加入对MXFX的荧光有静态猝灭作用,这种荧光静态猝灭作用是由ctDNA和MXFX结合引起的,作用力为氢键或范德华力,结合常数为1.28×10⁵ L/mol（25 ℃）。采用离子强度的影响、碘离子猝灭及溴乙锭竞争作用实验研究了MXFX与ctDNA间的相互作用,结果表明MXFX与DNA的结合是MXFX嵌入到DNA中相邻2个碱基对之间,属于嵌入结合模式。     

光谱法研究橙皮素与DNA分子间的相互作用机制

以盐酸小檗碱(BR)为荧光探针,研究了鲱鱼精DNA与橙皮素的相互作用.采用荧光光谱,紫外可见吸收光谱,盐效应,Scatchard方程等手段,探讨了橙皮素与DNA的作用机制.结果表明,在生理酸度(pH=7.4)下,橙皮素对DNA-BR体系的荧光有猝灭作用,猝灭类型为静态猝灭与动态猝灭共存的模式,其作用方式为嵌插和静电作用...     

靶向分子羧甲基多糖与DNA作用机理研究

以溴化乙锭(EB)为荧光探针,结合紫外光谱法研究了壳聚糖、羧甲基壳聚糖、羧甲基纤维素与DNA的相互作用,药物分子与DNA作用程度可用作用常数D来表示,实验表明: 几种生物多糖与DNA分子的作用强弱顺序为: 壳聚糖＞羧甲基壳聚糖＞羧甲基纤维素;多糖主要以嵌插方式与DNA作用, 使EB-DNA荧光猝灭。     

重金属Cu2+增强农药甲萘威对ctDNA作用的光谱法研究

运用荧光光谱和紫外吸收光谱研究了在Cu2 的参与下农药甲萘威与小牛胸腺DNA(ctDNA)的相互作用.实验结果表明,甲萘威可以嵌入小牛胸腺DNA的双链中形成DNA加合物,从而使甲萘威的荧光光谱发生猝灭.通过计算这种猝灭为静态猝灭.当有Cu2 的参与下,甲萘威的猝灭常数增大,结合位点数n也有很大的增加.通过对热力学函数的计算和分析,在有Cu2 参与时,金属离子可能在甲萘威分子与ctD-NA分子间起"离子架桥作用",使甲萘威分子与ctDNA分子间静电相互作用增强,故△H对△G的贡献增大.     

氨与牛血清白蛋白的相互作用

用荧光光谱法和紫外吸收光谱法研究了在生理pH值条件下氨与牛血清白蛋白(BSA)的相互作用,结果表明:氨对BSA的荧光有较强的猝灭作用,该猝灭属于同时具有动态猝灭和静态猝灭特征的联合猝灭,但以静态猝灭为主.根据猝灭结果求得了不同温度下氨与BSA相互作用的结合位点数、结合常数及反应热力学参数,并据此推测它们之间主要的相互作...     

Study on the Interaction of an Anthracycline Disaccharide with DNA by Spectroscopic Techniques and Molecular Modeling

This study was designed to examine the interaction of 4'-O-(a-L-Cladinosyl) daunorubicin (DNR-D5), a disaccharide anthracycline with calf thymus deoxyribonucleic acid (ctDNA) by UV/Vis in combination with fluorescence spectroscopy and molecular modeling techniques under physiological conditions (Britton-Robinson buffer solutions, pH?=?7.4). By the analysis of UV/Vis spectrum, it was observed that upon binding to ctDNA the anthraquinone chromophore of DNR-D5 could slide into the base pairs. Moreover, the large binding constant indicated DNR-D5 had a high affinity with ctDNA. At the same time, fluorescence spectra suggested that the quenching mechanism of the interaction of DNR-D5 to ctDNA was a static quenching type. The binding constants between DNR-D5 and ctDNA were calculated based on fluorescence quenching data at different temperatures. The negative ?G implied that the binding process was spontaneous, and negative ?H and negative ΔS suggested that hydrogen bonding force most likely played a major role in the binding of DNR-D5 to ctDNA. Moreover, the results obtained from molecular docking corroborate the experimental results obtained from spectroscopic investigations.     

染料木素铬(Ⅲ)配合物与DNA相互作用的研究

在pH 7.2 Tris缓冲溶液中,采用紫外、DNA熔点、荧光、粘度等手段研究了染料木素铬(Ⅲ)配合物与小牛胸腺DNA(ctDNA)的作用机制,探讨了其作用模式。当加入一定量的ctDNA时,染料木素铬(Ⅲ)配合物的紫外吸收光谱的最大吸收峰产生明显的减色效应;而配合物体系的荧光强度、共振光散射信号随ctDNA的加入逐渐增强。配合物的存在能有效猝灭EB-DNA体系的荧光,且猝灭方式为静态猝灭。同时ctDNA的熔点和ctDNA溶液的粘度随配合物的加入而增大。据此推断,染料木素铬(Ⅲ)配合物与ctDNA之间具有较强的作用,配合物主要以插入方式与DNA结合,二者的键合常数为1.9×105 L·mol-1。该研究结果提示染料木素铬(Ⅲ)配合物有望作为抗癌活性候选药物,值得进一步深入研究。     

Spectroscopic,Viscositic and Molecular Modeling Studies on the Interaction of 3′-Azido-Daunorubicin Thiosemicarbazone with DNA

A new daunorubicin has been synthesized and structurally characterized. The interaction of native calf thymus DNA (ctDNA) with 3′-azido-daunorubicin thiosemicarbazone (ADNRT) was investigated under simulated physiological conditions by multi-spectroscopic techniques, viscometric measurements and molecular modeling study. It concluded that ADNRT could intercalate into the base pairs of ctDNA, and the fluorescence quenching by ctDNA was static quenching type. Thermodynamic parameters calculated suggested that the binding of ADNRT to ctDNA was mainly driven by hydrophobic interactions. The relative viscosity of ctDNA increased with the addition of ADNRT, which confirmed the intercalation mode. Furthermore, molecular modeling studies corroborate the above experimental results.     

两种卤代荧光素与DNA作用荧光特性的研究

对四溴荧光素(TBF)、四氯四溴荧光素(TTF)两种卤代荧光素与DNA作用的荧光特性进行了研究,结果表明：TBF和TTF的最大λ_ex/λ_em为518/540 nm和540/560 nm,DNA的存在会使TBF和TTF的荧光强度发生变化;荧光猝灭实验和偏振实验表明：TBF与DNA的作用方式可能是沟槽键合和嵌插作用,而TTF 嵌插在了DNA碱基对之间;盐效应实验表明：溶液离子强度的大小会影响TBF和TTF与DNA的作用;荧光法测得TBF和TTF与DNA的结合常数为1×106和2×106 L·mol-1,结合位点数为0.62和0.16。     

染料木素及其葡萄糖苷与DNA相互作用的研究

在pH 7.2 Tris缓冲溶液中,采用紫外、荧光、粘度等方法研究了染料木素、染料木素葡萄糖苷、染料木素7,4’-二-O-β-D-葡萄糖苷与小牛胸腺ctDNA的作用。结果表明,在ctDNA存在下,染料木素及其葡萄糖苷的紫外吸收光谱的最大吸收峰均产生明显的减色效应。三种化合物的加入均能有效猝灭EB-DNA体系的荧光,而猝灭方式并不唯一。另外,ctDNA溶液的粘度也都随三种化合物的加入而增大。据此推断,染料木素及其葡萄糖苷与ctDNA之间具有较强的作用,可能以部分插入及氢键作用与ctDNA结合,其作用强弱顺序为染料木素葡萄糖苷>染料木素7,4’-二-O-β-D-葡萄糖苷>染料木素。结果提示染料木素的7位或4’位葡萄糖基化修饰有望作为抗癌活性候选物,值得进一步深入研究。     

Interaction Between Methylene Blue and CalfThymus Deoxyribonucleic Acid by Spectroscopic Technologies

Characterization of the interaction between methylene blue (MB) and calf thymus deoxyribonucleic acid (ctDNA) was investigated by UV absorption spectra, fluorescence spectra, fluorescence polarization and fluorescence quenching experiments by ferrocyanide. The above results indicated that the binding modes of MB to ctDNA were relative to the molar ratio γ (γ=[DNA]/[MB]). At low γ ratios (γ < 4), remarkable hypochromic effect with no shift of λ_max in the absorption spectra of MB was observed in the presence of increasing amounts of ctDNA, the fluorescence of MB was efficiently quenched by the ctDNA bases and the fluorescence polarization of MB was slightly increased, which indicated that MB cations bound to phosphate groups of ctDNA by electrostatic interaction and then stacked on the surface of ctDNA helix. While at high γ ratios (γ > 6), besides the fluorescence of MB was quenched efficiently by the ctDNA bases, a red shift (about 3 nm) in the absorption spectra of MB was observed and the fluorescence polarization of MB was obviously increased, which indicated the intercalation binding that MB molecules were intercalated into the space of two neighbouring DNA base pairs was the preferred mode. Effects of K₄Fe(CN)₆ on the fluorescence quenching of the MB-ctDNA system at low and high γ ratios were also performed. The results showed that at γ = 1.7, the quenching effect by ferrocyanide was higher than that of pure MB, while at γ = 13.6 a decreased quenching of the fluorescence intensity was observed as compared with that of pure MB, which further proved the above conclusion. In addition, the mechanisms of the hypochromic effect and the fluorescence quenching were also discussed in detail.     

Research on the interaction of cr(III) complex of genistein with DNA

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.     

光谱法及分子模拟研究青蒿素与小牛胸腺DNA的相互作用

在pH 7.4的Tris-HCl缓冲溶液中,采用紫外吸收光谱、荧光光谱结合溴化乙锭（EB）荧光探针、共振散射光谱以及DNA熔点（T_m）实验和分子模拟等技术,研究了青蒿素（QHS）与小牛胸腺DNA（ctDNA）分子间结合位点与结合机制。光谱实验结果显示,QHS与DNA发生减色效应,QHS的加入使EB-DNA体系发生静态荧光猝灭,QHS与DNA作用后其467 nm处共振散射峰锐增,与QHS作用引起DNA的T_m值升高5℃,说明QHS竞争性地嵌插入DNA的碱基对中。通过计算获得QHS与DNA间结合常数K_a为1.43×10³ L/mol（298 K）、0.99×10³ L/mol（304 K）。分子模拟结果表明,QHS吡喃环部分结构嵌插到DNA小沟区域GA碱基对间,氢键和范德华力是两者间结合的主要非共价作用方式,该结论与光谱法和热力学所得结果一致。