光谱及分子对接技术研究小分子药物与DNA、蛋白的相互作用

发布时间：2018-01-12 01:30

本文关键词：光谱及分子对接技术研究小分子药物与DNA、蛋白的相互作用　出处：《长春师范大学》2017年硕士论文　论文类型：学位论文

【摘要】：本文主要采用光谱法和分子对接法研究了几种小分子药物与血清白蛋白和DNA的相互作用,主要研究内容如下:在pH=7.40的条件下,采用多种光谱技术研究了隐丹参酮与人血清白蛋白(HSA)的相互作用。隐丹参酮对HSA的猝灭过程为静态猝灭,并获得了在不同温度下隐丹参酮与HSA作用的结合常数和结合位点数。热力学参数研究发现隐丹参酮与HSA的主要结合力为静电作用力。位点竞争实验结果表明隐丹参酮结合在HSA的site I位。同步荧光光谱法和圆二色光谱法的结果表明隐丹参酮的加入使HSA的二级结构发生变化。考察了金属离子对隐丹参酮-HSA体系的影响。根据F?rster理论,计算得到隐丹参酮与HSA之间的结合距离r0为2.86 nm。利用光谱法和分子对接法研究了昔萘酸沙美特罗与牛血清白蛋白(BSA)的相互作用。紫外光谱法表明BSA的加入对昔萘酸沙美特罗的紫外吸收光谱产生影响。荧光猝灭和时间分辨荧光光谱法的结果表明昔萘酸沙美特罗对BSA的猝灭过程为静态猝灭。得到了不同温度下的结合常数和结合位点数,结合作用力为静电作用。位点竞争实验和分子对接法的结果显示昔萘酸沙美特罗结合在BSA的site I位。红外光谱法、同步荧光光谱法和圆二色光谱法的结果表明昔萘酸沙美特罗的加入对BSA的二级结构产生影响。此外,还探究了K+、Ca2+、Cu2+、Zn2+和Fe3+对昔萘酸沙美特罗-BSA体系的影响。计算得到昔萘酸沙美特罗与BSA之间的结合距离为2.37 nm。采用光谱法、粘度法、电化学法和分子对接法研究氢溴酸右美沙芬与DNA的相互作用。荧光猝灭和时间分辨荧光光谱法的结果表明二者的猝灭过程为静态猝灭。获得了291、301和311 K下的结合常数和结合位点数,热力学参数的结果表明氢溴酸右美沙芬与DNA以范德华力或氢键相结合,结合距离为2.54 nm。红外光谱法和圆二色光谱法的结果表明氢溴酸右美沙芬的加入对DNA的二级结构产生了影响。紫外吸收光谱法、离子强度实验、粘度法、熔解温度实验和与单链DNA(ssDNA)作用的结果均表明氢溴酸右美沙芬与DNA的结合是嵌插作用。分子对接的结果表明二者以嵌插模式相结合,在结合的过程中,氢键发挥了重要作用。循环伏安法和微分脉冲伏安法的结果显示DNA的加入使氢溴酸右美沙芬的峰电位发生了移动。以吖啶橙(AO)为探针研究了硫酸新霉素和硫酸巴龙霉素与DNA的相互作用。荧光猝灭法和时间分辨荧光光谱法的结果表明硫酸新霉素/硫酸巴龙霉素与AO-DNA的作用是静态猝灭过程。在291 K时,硫酸新霉素和硫酸巴龙霉素与AO-DNA的结合常数分别为6.70×103和1.44×103 L·mol-1。ΔH、ΔS和ΔG的结果表明硫酸新霉素/硫酸巴龙霉素与AO-DNA以范德华力和氢键发生结合。红外光谱法和圆二色光谱法的结果表明硫酸新霉素/硫酸巴龙霉素的加入对DNA的二级结构产生了影响。离子强度实验、粘度法、熔解温度实验和与ssDNA作用的实验结果均证明硫酸新霉素/硫酸巴龙霉素与DNA以沟区模式相结合。分子对接法进一步表明硫酸新霉素/硫酸巴龙霉素结合在DNA小沟区的A-T富裕区。采用光谱法、粘度法和分子对接法研究了以溴化乙锭(EB)为荧光探针小诺霉素/妥布霉素与DNA的相互作用。荧光猝灭法和时间分辨荧光光谱法的结果表明小诺霉素/妥布霉素与EB-DNA的作用是静态猝灭过程。得到了不同温度下小诺霉素/妥布霉素与EB-DNA的结合常数和结合位点数,ΔH、ΔS和ΔG的结果表明小诺霉素/妥布霉素与EB-DNA以范德华力和氢键发生结合。红外光谱法和圆二色光谱法表明小诺霉素/妥布霉素的加入对DNA的二级结构产生了影响。离子强度实验、粘度法、熔解温度实验和单双链DNA对比实验的结果均表明小诺霉素/妥布霉素与DNA的作用模式为沟区结合。分子对接法表明小诺霉素/妥布霉素与DNA以沟区模式相结合,在结合过程中氢键发挥了重要作用。
[Abstract]:This paper mainly uses the interaction of spectroscopy and molecular docking method of several small molecule drugs and serum albumin and DNA, the main research contents are as follows: under the condition of pH=7.40 of cryptotanshinone and human serum albumin (HSA) using a variety of spectroscopic techniques. The interactions of cryptotanshinone on quenching process for HSA static quenching, and the binding constants obtained at different temperatures of cryptotanshinone and HSA and the number of binding sites. The study found the main thermodynamic parameters of the binding force of cryptotanshinone and HSA electrostatic interaction. Experimental results show that the site competition with Cryptotanshinone in HSA site I. Synchronous fluorescence spectroscopy and circular two color spectroscopy results showed cryptotanshinone with two level structure makes the change of HSA. The effects of metal ions on the cryptotanshinone -HSA system. According to the F? Rster theory, calculation of cryptotanshinone With the combination of HSA and the distance between the R0 is studied with naphthalene Shah Mette Lo acid and bovine serum albumin in 2.86 nm. by spectroscopy and molecular docking method (BSA). The interactions of UV spectra showed that the addition of BSA on UV in naphthalene acid Shah Mette Lo had influenced the absorption spectra. The fluorescence quenching and time-resolved fluorescence spectroscopy the results show that with naphthalene acid Shah Mette Lo BSA quenching is static quenching. The binding constants and binding sites under different temperatures, binding force is electrostatic interaction. The results of site competition experiments and molecular docking method show that with naphthalene acid sand Emmett combination in BSA site, I. Infrared spectroscopy, synchronous fluorescence round two color spectroscopy and spectrometry results show that with naphthalene acid Shah Mette Lo joined the two stage structure influence BSA. In addition, it also explores the K+, Ca2+, Cu2+, Zn2+ and Fe3+ in Shah Mette Lo -BSA of naphthalene acid The effects obtained. Combined with the distance of 2.37 nm. by spectroscopy, viscosity method between naphthalene acid and BSA in Shah Mette Lo, the interaction of electrochemical method and molecular docking method of Dextromethorphan Hydrobromide and DNA. Fluorescence quenching and time-resolved fluorescence spectroscopy results show that the quenching process of the two is a static quenching procedure. The binding constants and binding sites of 291301 and 311 K, the results show that the thermodynamic parameters of Dextromethorphan Hydrobromide combined with DNA by Fan Dehua force or hydrogen bonding, the binding distance is 2.54 nm. the results of infrared spectroscopy and circular dichroism method showed that two Dextromethorphan Hydrobromide added two level structure of the influence of DNA. UV absorption spectroscopy, ionic strength experiments, viscosity, melting temperature and experiment with single stranded DNA (ssDNA) results show that the combined effect of Dextromethorphan Hydrobromide and DNA is intercalation. The docking results showed that the two combination in the mode of intercalation, in the process of combining hydrogen bonds play an important role. Cyclic voltammetry and differential pulse voltammetry. The results showed that DNA was added to make the peak potential of Dextromethorphan Hydrobromide shifted. Using acridine orange (AO) as a probe of neomycin sulfate and by interaction of Barone sulfate peptide and DNA. Fluorescence quenching and time-resolved fluorescence spectroscopy results showed that neomycin sulfate / sulfuric acid azithromycin and Barone AO-DNA is a static quenching process. In 291 K, neomycin sulfate and sulfuric acid by Barone and AO-DNA binding constants were 6.70 * 103 and 1.44 * 103 L - mol-1. Delta H, Delta S and Delta G showed that neomycin sulfate / Barone sulfate kanamycin and AO-DNA by van Edward force and hydrogen bonding. Infrared spectroscopy and circular two color spectrometry results show that neomycin sulfate sulfate / Barone Two adding the lignin structure of DNA was influenced. The ionic strength experiments, viscosity, melting temperature and ssDNA effect experiment and experimental results show that neomycin sulfate / Barone sulfate kanamycin and DNA in groove pattern combining. Molecular docking method further showed that neomycin sulfate sulfur acid / Barone zymoid combination in DNA groove A-T rich area. By using spectroscopy, viscosity and molecular docking method was studied by using ethidium bromide (EB) as a fluorescence probe interaction of micronomicin / tobramycin and DNA. Fluorescence quenching and time-resolved fluorescence spectroscopy results show that micronomicin / tobramycin and the role of EB-DNA is static quenching at different temperatures are obtained. The process of micronomicin / tobramycin and EB-DNA binding constants and binding sites, Delta H, Delta S and delta G results indicated that micronomicin / EB-DNA and tobramycin by van Edward force and hydrogen bond. . infrared spectroscopy and circular dichroism method showed that two micronomicin / tobramycin adding two structure of DNA was influenced. The ionic strength experiments, viscosity, melting temperature and experiment of single stranded DNA experimental results showed that the micronomicin / role model and DNA for groove tobramycin with the show. The molecular docking method of micronomicin / tobramycin and DNA in groove combination mode, play an important role in the process of combining hydrogen bonds.

【学位授予单位】：长春师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R96;O657.3

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