Mechanistic study for porpyrin metallation occurring in DNA-quartet structure

DNA四重体结构中卟啉金属化的机理研究

• 批准号: 11640563
• 负责人: TAJIMA Kunihiko
• 金额: $ 2.3万
• 依托单位: Kyoto Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11640563/
• 关键词: DNA; DNA-enzyme; heme; DNA-heme interaction; heme-metallation; 銅錯体; ポルフィリン; 四重鎖構造; G-カルテット; 金属挿入反応; 核磁気共鳴法; 円二色性分光

In this study, the structural study on porphyrin-DNA interactions was performed in order to clarify whether it is possible for single-stranded DNAs to fold to create specific binding pockets for anionic porphyrins, which do not naturally bind to nucleic acids.The kinetics of the metallation of Cu^<2+> into MPIX was first examined in the presence and absence of PS5. ST1 (dTCGTGGGTCATTGTGGGTGGGTGTGGCTGGTCC) and other G-rich oligomers by HPLC.The results were almost identical to that as previously reported (3).CD spectroscopy has been used to distinguish between parallel and antiparallel G-quadruplexes (4, 5) The former quadruplexes exhibit a positive CD band around 260 nm and a negative CD band at 240 nm. The latter quadruplexes exhibit a positive CD band around 290 nm and a weaker negative band at 265 nm. CD spectroscopy was carried out for PS5. ST1 (25 μM) at various temperatures in the absence and presence of K^+ ion. In the absence of K^+, this sample showed two positive peaks at 265 and 285 nm at 0 ℃, while at increasing temperatures, one at 265 nm disappeared, and the other at 285 nm decreased. Temperature-dependent CD spectroscopy was also performed for PS5. ST 1 in the presence of K^+ ion in the temperature range of 0-80 ℃. Both the positive peaks appeared below 25 ℃, and the intensity of the peak at 265 nm was much stronger than that in the absence of K^+ ion. These results indicate that even in the absence of K^+ ion, PS5. ST 1 takes an ordered structure, and that the addition of K^+ ion induces components of parallel G-quadruplexes.^1H NMR measurements were carried out for PS5. ST 1 in the absence and presence of K^+ ion. The iminoproton signals are an indicator of symmetry and stoichiometry. The NMR spectra show some change in the chemical shifts of the peaks, which may correspond with the change in the CD spectra.

在这项研究中，对卟啉-DNA相互作用的结构进行了研究，以澄清单链DNA是否有可能折叠以产生特异性结合阴离子卟啉的口袋，阴离子卟啉不与核酸结合。ST1（dTCGTGGGTCATTGTGGGTGGGTGTGGCTGGTCC）和其它富G寡聚体的HPLC分析结果与文献报道的结果基本一致（3）. CD光谱用于区分平行和反平行G-四链体（4，5）.后者四链体在290 nm附近显示正CD带，在265 nm处显示较弱的负带。对PS5进行CD光谱分析。ST 1（25 μ M）在不存在和存在K ^+离子的情况下在不同温度下。在不存在K ^+的情况下，该样品在0 ℃时在265和285 nm处显示出两个正峰，而随着温度的升高，265 nm处的一个峰消失，285 nm处的另一个峰减少。还对PS5进行了温度依赖性CD光谱。ST 1在0 - 80 ℃温度范围内，K ^+离子存在时，晶体结构稳定。两个正峰均出现在25 ℃以下，且在265 nm处的峰强度远强于无K ^+存在时的峰强度。这些结果表明，即使在没有K ^+离子的情况下，PS5. ST 1具有有序结构，K ^+离子的加入诱导了平行G-四链体的成分。对PS5进行1H NMR测量。ST 1在不存在和存在K ^+离子的情况下。亚氨基质子信号是对称性和化学计量的指示剂。NMR谱显示出峰的化学位移的一些变化，这可能与CD谱的变化相对应。

项目成果

S.Oka, K.Tajima, and H,Sakurai: "A Chemical Model of Tryptophan 2, 3-Dioxygenase : Reactivity and Formation of a Reactive Intermadiate in the Dioxygenase"Chem. Pharm. Bull.. 46. 377-383 (1991)

S.Oka、K.Tajima 和 H，Sakurai：“色氨酸 2, 3-双加氧酶的化学模型：双加氧酶中反应性中间体的反应性和形成”Chem。

Kenji Kanaori: "Structure and stability of consecutive stereoregulated chiral phosphorothioate DNA duplex"Biochemistry. 38. 16058-16066 (1999)

Kenji Kanaori：“连续立体调节手性硫代磷酸酯 DNA 双链体的结构和稳定性”生物化学。

Yuji Kajikawa: "Crystallographic and ESR Studies of π-π Stacked Binuclear Copper(II) Complexes of 1,3-Bis(quinoline-2-carboxamido) and 1,3-Bis(isoquinoline-3-carboxamido) propane,"Inorganica Chimica Acta. 288. 90-100 (1999)

Yuji Kajikawa：“1,3-Bis(quinoline-2-carboxamido) 和 1,3-Bis(isoquinoline-3-carboxamido) propane π-π Stacked Binuclear Copper(II) Complexes 的晶体学和 ESR 研究，”Inorganica Chimica法。288。90-100（1999）

Yuji Kajikawa: "Crystallographic and ESR Studies of π-π Stacked Binuclear Copper (II) Complexes of 1,3-Bis (quinoline-2-carboxamido) and 1,3-Bis (isoquinoline-3-carboxamido) propane,"Inorganica Chimica Acta. 288. 90-100 (1999)

Yuji Kajikawa：“1,3-Bis（喹啉-2-甲酰胺基）和 1,3-Bis（异喹啉-3-甲酰胺基）丙烷的 π-π 堆叠双核铜 (II) 配合物的晶体学和 ESR 研究，”Inorganica Chimica法。288。90-100（1999）

Yuji Kajikawa: "Crystallographic and ESR studies of π-π stacked binuclear copper (II) complexes of bis (quinoline-3-caeboxiamido) propane"Inorg.Chimica.Acta. 288. 90-100 (1999)

Yuji Kajikawa：“双（喹啉-3-羧酰胺基）丙烷的 π-π 堆叠双核铜 (II) 配合物的晶体学和 ESR 研究”Inorg.Chimica.Acta 288. 90-100 (1999)。