莱克多巴胺与小牛胸腺DNA的沟槽结合模式研究
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- 英文论文题名：Groove binding mode of ractopamine with calf thymus DNA
- 论文作者：夏开心 ; 张国文
- 英文论文作者：Xia Kaixin ; Zhang Guowen ; State Key Laboratory of Food and Technology ; Nanchang University
- 年：2016
- 作者机构：南昌大学食品科学与技术国家重点实验室;
- 论文关键词：莱克多巴胺 ; 小牛胸腺DNA ; 沟槽结合 ; 光谱法 ; 分子模拟
- 英文论文关键词：ractopamine ; calf thymus DNA ; groove binding ; spectroscopy ; molecular modeling
- 会议召开时间：2016-11-09
- 会议录名称：中国食品科学技术学会第十三届年会论文摘要集
- 英文会议录名称：Abstracts of the 13th Annual Meeting of CIFST
- 语种：中文
- 分类号：Q75
- 学会代码：ZGSP
- 会议名称：中国食品科学技术学会第十三届年会
- 会议地点：中国北京
- 主办单位：中国食品科学技术学会
- 学会名称：中国食品科学技术学会
- 页数：2
- 文件大小：114k
- 原文格式：O
- 会议级别：全国
摘要
在生理酸度(pH 7.4)的条件下,构建了基于化学计量学多元曲线分辨-交替最小二乘法(MCR-ALS)结合紫外-可见光谱、荧光光谱、圆二色谱(CD)、红外光谱(FT-IR)以及分子模拟和熔点、粘度测量的方法体系,研究了莱克多巴胺(RAC)与小牛胸腺DNA(ctDNA)的相互作用。结果表明,ctDNA以静态方法猝灭RAC的内源荧光,25℃时ctDNA与RAC的结合常数为1.45×10~4 L·mol~(-1),氢键和范德华力是两者结合的主要驱动力。运用MCR-ALS方法解析RAC与ctDNA反应体系的紫外扩展矩阵,获得反应组分(RAC、ctD NA和RAC-ctDNA复合物)的浓度变化和纯组分的光谱曲线,实现了RAC-ctDNA相互作用的定量监测。盐效应、单双链实验、碘化钾猝灭实验、ctDNA熔点和粘度测量及FT-IR分析结果表明,RAC结合在ctDNA的小沟富集区,且主要与胸腺嘧啶发生作用,分子模拟证实这一实验结果。圆二色谱结果显示,RAC与胸腺嘧啶作用引起ctDNA的B构象向A构象转变。该研究为深入认识RAC与ctDNA的作用机制以及RAC的毒理行为提供了理论依据
The interaction of ractopamine with calf thymus DNA(ctDNA) was investigated in physiological buffer(pH 7.4)by the multivariate curve resolution-alternating(MCR-ALS) chemometrics algorithm combined with UV-vis absorption,fluorescence,circular dichroism(CD) and Fourier transform infrared(FT-IR) spectroscopy as well as molecular simulation,melting and viscosity measurements.The results showed that ctDNA could quench the intrinsic fluorescence of RAC by static quenching.The binding constant between RAC and ctDNA was calculated to be 1.45 × 10~4 L·mol~(-1) at 25℃,and the binding of RAC to ctDNA was driven mainly by hydrogen bonding and van der Waals interaction.The MCR-ALS analysis extracted simultaneously the concentration profiles and spectra for the three components(RAC,ctDNA and RAC-ctDNA complex) to quantitatively monitor the RAC-ctDNA interaction.The binding mode of RAC to ctDNA was principally through groove binding and RAC preferentially bound to thymine base of ctDNA,as evidenced by ctDNA melting temperature studies,viscosity measurements,salt effect,iodide quenching effect,single stranded ctDNA and double stranded ctDNA quenching experiments,and FT-IR analysis,these results were further confirmed by molecular simulation.The CD spectra indicated that the binding of RAC to thymine base led to a transformation B-like DNA structure to A-like conformation.This study has provided insights into the binding mechanism of ractopamine with ctDNA and toxicologic behavior of the compound.
The interaction of ractopamine with calf thymus DNA(ctDNA) was investigated in physiological buffer(pH 7.4)by the multivariate curve resolution-alternating(MCR-ALS) chemometrics algorithm combined with UV-vis absorption,fluorescence,circular dichroism(CD) and Fourier transform infrared(FT-IR) spectroscopy as well as molecular simulation,melting and viscosity measurements.The results showed that ctDNA could quench the intrinsic fluorescence of RAC by static quenching.The binding constant between RAC and ctDNA was calculated to be 1.45 × 10~4 L·mol~(-1) at 25℃,and the binding of RAC to ctDNA was driven mainly by hydrogen bonding and van der Waals interaction.The MCR-ALS analysis extracted simultaneously the concentration profiles and spectra for the three components(RAC,ctDNA and RAC-ctDNA complex) to quantitatively monitor the RAC-ctDNA interaction.The binding mode of RAC to ctDNA was principally through groove binding and RAC preferentially bound to thymine base of ctDNA,as evidenced by ctDNA melting temperature studies,viscosity measurements,salt effect,iodide quenching effect,single stranded ctDNA and double stranded ctDNA quenching experiments,and FT-IR analysis,these results were further confirmed by molecular simulation.The CD spectra indicated that the binding of RAC to thymine base led to a transformation B-like DNA structure to A-like conformation.This study has provided insights into the binding mechanism of ractopamine with ctDNA and toxicologic behavior of the compound.
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