Detection of DNA structural changes upon drug-binging by a combination of site-selective deutaretation and difference Raman intensity changes

通过结合位点选择性氘化和差异拉曼强度变化来检测药物结合时 DNA 结构的变化

• 批准号: 15590036
• 负责人: TOYAMA Akira
• 金额: $ 1.92万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590036/
• 关键词: UV resonance Raman spectroscopy; Drug-nucleic acid interaction; Site-selective deuterated DNA; Nucleic acid structure; 共鳴ラマン分光; DNA-薬物相互作用; 核酸-薬物相互作用

The goal of my research is to understand the mechanism of drug-DNA interaction by using of"isotope-edited UV resonance Raman spectroscopy -a combination of site-selective isotope labeling and ultraviolet resonance Raman (UVRR) spectroscopy The aim of this project is to find the correlation between the structure and UVRR intensity of DNA bases.UV Raman spectra of adenine and guanine derivatives were recorded in several solvents and the intensities of UV Raman bands were analyzed by using three solvent parameters proposed by Kamlet and Taft : the hydrogen-bond donor acidy, hydrogen-bond acceptor basicity, and polarity-polarizability. As a result, we have found that the Raman intensity of the purine rings depends mostly on hydrogen bonding but little on environmental polarity-polarizability.However, it is also known that base stacking interactions suppress the Raman intensity of DNA bases (Raman hypochromism). In order to determine either hydrogen-bonding or stacking is dominant in Raman … More intensity, isotope-edited UVRR spectra were measured of a 22-mer oligonucleotide (LacDNA) bound by cyclic AMP (cAMP) receptor protein (CRP). CRP-(cAMP)2 complex forms strong hydrogen bonds with guanine residues of LacDNA, but base stacking of the guanine rings are not significantly altered by the complex. For isotope-edited UVRR spectra of the guanine residues of LacDNA, some Raman bands showed appreciable frequency changes but did not show meaningful intensity changes upon CRP-(cAMP)2 binding. It is indicated that the UVRR intensities of aqueous DNA must predominantly be determined by the strength of the base stacking. Isotope edited UVRR spectra of a 19-mer oligonucleotide containing a AC-mismatched base pair were also measured to evaluate the sensitivity of the UVRR intensity to the base stacking. A significant intensity change was detected at the pre-melting state of the DNA. This observation demonstrates the utility of isotope-edited UVRR spectroscopy as a probe of the base staking of DNA bases. Less

本研究的目的是利用“同位素编辑紫外共振拉曼光谱--一种结合位点选择性同位素标记和紫外共振拉曼（UVRR）的方法”来了解药物与DNA相互作用的机制。本课题的目的是寻找DNA碱基的结构与UVRR强度之间的相关性，记录了腺嘌呤和鸟嘌呤衍生物在几种溶剂中的紫外拉曼光谱，并计算了它们的UVRR强度。采用Kamlet和塔夫脱提出的三个溶剂参数：氢键给体酸性、氢键受体碱性和极性-极化率对紫外拉曼谱带进行了分析。结果表明，嘌呤环的拉曼强度主要取决于氢键，而与环境极性-极化率关系不大，但碱基堆积作用抑制了DNA碱基的拉曼强度（拉曼减色效应）。为了确定在拉曼光谱中是氢键还是堆积起主导作用， ...更多信息 强度，测量由环AMP（cAMP）受体蛋白（CRP）结合的22-mer寡核苷酸（LacDNA）的同位素编辑的UVRR光谱。CRP-（cAMP）2复合物与LacDNA的鸟嘌呤残基形成强氢键，但复合物不显著改变鸟嘌呤环的碱基堆积。对于LacDNA鸟嘌呤残基的同位素编辑的UVRR光谱，一些拉曼带显示出明显的频率变化，但在CRP-（cAMP）2结合后没有显示出有意义的强度变化。结果表明，DNA水溶液的UVRR强度主要取决于碱基堆积的强度。还测量了含有AC错配碱基对的19-mer寡核苷酸的同位素编辑的UVRR光谱，以评估UVRR强度对碱基堆积的灵敏度。在DNA的预解链状态下检测到显著的强度变化。这一观察结果证明了同位素编辑的UVRR光谱作为DNA碱基的碱基固定的探针的实用性。少

项目成果

Correlation between UV Raman intensity and hydrogen bonding of the adenine ring

紫外拉曼强度与腺嘌呤环氢键的相关性

• 发表时间: 2005
• 期刊: Journal of Molecular Structure 735-736
• 作者: T.Honda;H.Namiki;H.Nagase;H.Mizutani;Akira Toyama
• 通讯作者: Akira Toyama