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苯菲啶生物碱 nitidine 将低 pH 值、低温诱导的质子化 DNA 形式结构改变为规范形式:光谱研究。

Structural alteration of low pH, low temperature induced protonated form of DNA to the canonical form by the benzophenanthridine alkaloid nitidine: Spectroscopic exploration.

机构信息

Physical Chemistry Section, Department of Chemistry, Jadavpur University, Raja S. C. Mullick Road, Jadavpur, Kolkata 700 032, India.

Physical Chemistry Section, Department of Chemistry, Jadavpur University, Raja S. C. Mullick Road, Jadavpur, Kolkata 700 032, India.

出版信息

Int J Biol Macromol. 2018 Nov;119:1106-1112. doi: 10.1016/j.ijbiomac.2018.08.031. Epub 2018 Aug 8.

DOI:10.1016/j.ijbiomac.2018.08.031
PMID:30098360
Abstract

Polymorphism of DNA plays a very important part of research relating to the drug-DNA interactions. Here main focus of our investigation is to monitor the interaction of the benzophenanthridine plant alkaloid, nitidine (NIT) with two different forms of DNA i.e. B-DNA and protonated form of DNA maintaining proper temperatures and buffer conditions. Binding interaction of NIT was ascertained from the UV-Visible spectroscopic and spectrofluorimetric titration experiments. Binding constants of the interactions of NIT with different polymorphic forms were calculated from UV-absorption study. The binding constants were 3.8 × 10 M and 1.3 × 10 M for B-DNA and protonated DNA respectively. Red shift in the absorption maxima of NIT on binding with DNA, comparatively greater relative quenching of fluorescence intensity of free NIT than bound NIT, perturbation in the CD spectrum of DNA in presence of NIT confirmed the mode of binding as intercalation. Moreover, spectropolarimetric experiment confirms that left handed protonated form of DNA gets partially converted to the canonical B form of DNA while binds with NIT. Besides the CD experiment, thermal melting experiment also showed that on binding with NIT stabilization of protonated DNA was increased to an appreciable extent.

摘要

DNA 的多态性在与药物-DNA 相互作用相关的研究中起着非常重要的作用。我们的研究重点主要是监测苯并菲啶植物生物碱氮啶(NIT)与两种不同形式的 DNA 即 B-DNA 和质子化 DNA 在适当温度和缓冲条件下的相互作用。通过紫外可见光谱和荧光光谱滴定实验确定了 NIT 的结合相互作用。从紫外吸收研究中计算了 NIT 与不同多态形式相互作用的结合常数。B-DNA 和质子化 DNA 的结合常数分别为 3.8×10 M 和 1.3×10 M。NIT 与 DNA 结合时吸收最大值的红移、自由 NIT 荧光强度的相对猝灭程度明显大于结合 NIT、NIT 存在时 DNA 的 CD 光谱的扰动均证实了结合模式为嵌入。此外,旋光光谱实验证实,与 NIT 结合时,左手质子化形式的 DNA 部分转化为典型的 B 形式 DNA。除了 CD 实验,热融实验还表明,与 NIT 结合后,质子化 DNA 的稳定性得到了相当大的提高。

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