Detection of small structural changes of DNA under physiological conditions by a combination of site-selective deuteration and Raman difference spectroscopy

结合位点选择性氘化和拉曼差异光谱检测生理条件下 DNA 的微小结构变化

• 批准号: 13672248
• 负责人: TOYAMA Akira
• 金额: $ 2.24万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672248/
• 关键词: site-selective deuterated DNA; resonance Raman spectroscopy; nucleic acid structure; nucleic acid-drug interaction

Small structural changes of selected guanine residues in DNA have been detected by a combination of site-selective isotope labeling and ultraviolet resonance Raman (UVRR) spectroscopy, I.e. isotope-edited Raman spectroscopy. UVRR spectra were measured for [8-^2H]guaninelabeled and unlabeled oligonucleotides excited at 251nm light. In the difference spectrum between the unlabeled and labeled DNA, Raman signals arise from the residue at the labeled position and the other components were canceled out. Thus, it becomes possible to extract the Raman signals of the selected guanine in olinonucleotides.To obtain basic data for applying isotope-edited Raman spectroscopy to guanine residues, vibrational modes of UVRR bands of the C8-deuterated guanine ring were studied by examining the wavenumber shifts upon seven isotopic substitutions (2-^<13>C, 2-^<15>N, 6-^<18>O, 7-^<15>N, 8-^<13>C, 9-^<15>N and 1'-^<13>C). The hydrogen bond sensitivities of the Raman bands were also investigated by compari … More ng the Raman spectra recorded in several solvents of different hydrogen bonding properties. Some of the Raman bands have been found to be hydrogen bonding markers at specific donor or acceptor sites on the guanine ring, A negative peak around 1525 cm^<-1> a strong positive/negative peak pair around 1485/1465 cm^<-1>, and a weak positive/negative peak pair around 1025/1040cm^<-1> serve as markers of hydrogen bonding at N7, C6=O, and N1-H/C2-NH_2, respectively.The applicability of isotopeedited Raman spectroscopy has been tested by using a 22-mer oligonucleotide duplex. UVRR spectra of the labeled and unlabeled oligomers were measured in the absence and presence of cAMP receptor protein (CRP),Which is Known to hydrogen-bonded to guanine residues of duplex DNA. The isotope-edited spectra showed that CRP is hydrogen-bonded strongly to C6=O of guanine residues but not N7 atom. Isotope-edited Raman spectroscopy can provide structural information of a selected guanine residue for DNA-drug systems under physiological conditions. Less

通过位点选择性同位素标记和紫外线共振拉曼（UVRR）光谱（即同位素编辑拉曼光谱）的结合，可以检测到DNA中选定的鸟嘌呤残基的微小结构变化。在251nm光激发下，测量了[8-^2H]鸟嘌呤标记和未标记的寡核苷酸的UVRR光谱。在未标记和标记DNA的差谱中，拉曼信号来自标记位置的残基，其他成分被抵消。因此，可以提取所选鸟嘌呤的拉曼信号。为了获得将同位素编辑拉曼光谱应用于鸟嘌呤残基的基础数据，通过检测7个同位素取代（2-^<13>C, 2-^<15>N, 6-^<18>O, 7-^<15>N, 8-^<13>C, 9-^<15>N和1'-^<13>C）的波数移位，研究了c8 -氘化鸟嘌呤环UVRR波段的振动模式。通过比较几种不同氢键性质的溶剂中记录的拉曼光谱，研究了拉曼带的氢键灵敏度。在鸟嘌呤环的特定供体或受体位置上发现了一些拉曼带作为氢键标记，在1525 cm^<-1>附近有一个负峰，在1485/1465 cm^<-1>附近有一个强的正/负峰对，在1025/1040cm^<-1>附近有一个弱的正/负峰对，分别作为N7、C6=O和N1-H/C2-NH_2的氢键标记。用22聚寡核苷酸双链测试了同位素编辑拉曼光谱的适用性。在没有和存在cAMP受体蛋白（CRP）的情况下，测量了标记和未标记的低聚物的UVRR光谱，该蛋白已知与双链DNA的鸟嘌呤残基氢键结合。结果表明，CRP与鸟嘌呤残基的C6=O原子呈强氢键关系，而与N7原子无强氢键关系。同位素编辑拉曼光谱可以为生理条件下的dna -药物系统提供选定的鸟嘌呤残基的结构信息。少

项目成果

Akira Toyama: "Raman spectra and normal coordinate analysis of the N1-H and N3-H tautomers of 4-methylimidazole :"Journal of Physical Chemistry A. 106. 3403-3412 (2002)

Akira Toyama：“4-甲基咪唑的 N1-H 和 N3-H 互变异构体的拉曼光谱和正态坐标分析：”物理化学杂志 A. 106. 3403-3412 (2002)

Akira Toyama: "Assignments and hydrogen bonding sensitivity of UV resonance Raman bands of C8-deuterated guanine ring"Journal of Raman Spectroscopy. 33. 699-708 (2002)

Akira Toyama：“C8-氘化鸟嘌呤环的紫外共振拉曼带的归属和氢键敏感性”拉曼光谱杂志。

Akira Toyama: "Raman spectra and normal coordinate analysis of the N1-H and N3-H tautomers of 4-methylimidazole"The Journal of Physical Chemistry. (発表予定).

Akira Toyama：“4-甲基咪唑的 N1-H 和 N3-H 互变异构体的拉曼光谱和正态坐标分析”《物理化学杂志》（待出版）。

Ajdra Toyama: "Raman spectra and normal coordinate analysis of the N1-H and N3-H tautomers of 4-methylimidazole"Journal of Physical Chemistry A. 106. 3403-3412 (2002)

Ajdra Toyama：“4-甲基咪唑的 N1-H 和 N3-H 互变异构体的拉曼光谱和正态坐标分析”物理化学杂志 A. 106. 3403-3412 (2002)

Akira Toyama: "Raman spectra and normal coordinate analysis of the N1-H and N3-H tautomers of 4-methylimidazole"The Journal of Physical Chemistry A. 106・14. 3403-3412 (2002)

Akira Toyama：“4-甲基咪唑的N1-H和N3-H互变异构体的拉曼光谱和正态坐标分析”物理化学杂志A. 106・14（2002年）。