STUDY OF HYDROLYTIC ACTIVITIES OF THREE COMPONENT COMPLEXES OF DNA・LIGANDS・METAL ION

DNA·配体·金属离子三组分配合物的水解活性研究

• 批准号: 07680635
• 负责人: OGINO Kenji
• 金额: $ 1.47万
• 依托单位: OSAKA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07680635/
• 关键词: PLASMIDE DNA; HYDROLYSIS; METAL ION COMPLEX; RESTRICTION ENZYME MODEL

Metal ion effects on the phosphoryl and nucleotidyl hydrolyses and transfer reactions have been a long lasting subject of broad interests. We studied the site-selective hydrolyses of UTPs (ATP and UTP) promoted by metallo-complexes of appropriate ligands. The ligands (L) synthesized and examined were 2,2'-dipyridylamine (dpa) derivatives in which dpa is linked to adenine, thymine, and guanine mojities with a methylene-spacer of different lengths.The hydrolyses of ATP and UTP were carried out at pH 7.3 and 40ﾟC for 24 h and the amounts of the hydrolyses (NDP and NMP) were monitored by an anion-exchange HPLC.The 1 : 1 stoichiometric complexes of Cu(L)^<2+>, adenine containing ligand, were found to be superior in both the activities (hydrolyses %) and site-selectivities (ratios of ADP/AMP and UDP/UMP) as compared to other divalent metal ions (Cu^<2+>, Mg^<2+>, Zn^<2+> and Ni^<2+>) alone or their M (L) ^<2+>-complexes. The Cu(L)^<2+>-complexes showed the preferential cleavages of the terminal gamma-phosphate groups, although Cu(dpa)^<2+>, the active site of these ligands, inhibited their hydrolyses. The effect of reactivities of these complexes and their spacer-lengths suggests the interactions of the base-base moieties between ATP or UTP and the ligands. The formation of the ternary complex of Zn^<2+> : L : ATP was studies by using ^1H-and ^<31>P- NMR spectra. The hydrolytic activities of other ligands are investigated now.As DNA cleaving reagents, these M^<2+>-ligand complexes were examined to investigate their reactivities for thecleavages of plasmide DNA (pCU 18) and the Zn(L)^<2+> complexes, ademine containing ligand, were found to be effective (supercoil plasmide DNA (Form I) hydrolyzed to nicked (Form II) and linear DNA (Form III), measured after electrophyrisis) and indicated to work as a kind of models of distriction enzymes.

金属离子对磷酰基和核苷酸基水解和转移反应的影响一直是人们广泛关注的一个长期课题，我们研究了适当配体的金属配合物促进的UTP（ATP和UTP）的位点选择性水解。合成并检查了 2,2'-二吡啶胺 (dpa) 衍生物，其中 dpa 与腺嘌呤、胸腺嘧啶和ATP和UTP的水解在pH 7.3和40℃下进行24小时，并通过阴离子交换HPLC监测水解物（NDP和NMP）的量。发现含有腺嘌呤的配体 Cu(L)^2+ 的 1:1 化学计量配合物在与其他二价金属离子（Cu^<2+>、Mg^<2+>、Zn^<2+> 和 Ni 相比）的活性（水解%）和位点选择性（ADP/AMP 和 UDP/UMP 的比率） ^<2+>)单独或其M(L)^<2+>-复合物Cu(L)^<2+>-复合物显示末端的优先裂解。 γ-磷酸基团，尽管这些配体的活性位点Cu(dpa)^2+抑制了它们的水解。这些复合物的反应性及其间隔基长度的影响表明了ATP之间碱基-碱基部分的相互作用。或UTP和配体Zn^2+:L:ATP的三元复合物的形成通过使用^1H-和^31P-NMR光谱进行研究。现在研究其他配体的水解活性。作为DNA切割试剂，这些M^<2+>-配体复合物被检查以研究它们对质粒DNA(pCU 18)和Zn(L)^<2+的切割的反应性> 含有配体的腺胺复合物被发现是有效的（超螺旋质粒 DNA（I 型）水解为带切口（II 型）和线性 DNA（III 型），电熔后测量）并表明其可作为一种区域酶模型。

项目成果

M.Sawa, K.Ogino, and W.Tagaki: "Reactivities and Site-Selectivities of Hydrolysis of ATP and UTP Promoted by Metal Complexes of Adenine-Linked Di-2-pyridylamine Derivatives" Bull.Chem.Soc.Jpn.69. 765-774 (1996)

M.Sawa、K.Ogino 和 W.Tagaki：“腺嘌呤连接的二-2-吡啶胺衍生物金属配合物促进的 ATP 和 UTP 水解的反应性和位点选择性”Bull.Chem.Soc.Jpn.69。

Masaaki Sawa: "Reactivities and Site-Selectivities of Hydrolyses of ATP and UTP Promoted by Metal Complexes of Adenine-linked Di-2-pyridylamine Derivatives" Bull.Chem.Soc.Jpn.69. 765-774 (1996)

Masaaki Sawa：“腺嘌呤连接的二-2-吡啶胺衍生物金属配合物促进的 ATP 和 UTP 水解的反应性和位点选择性”Bull.Chem.Soc.Jpn.69。

Masaaki SAWA: "Reactivities and Site-Selectivities of Hydrolyses of ATP and UTP Promoted by Metal Complexes of Adenine-linked Di-2-pyridylamine Derivatives" Bull.Chem.Soc.Jpn.69 (3). 765-774 (1996)

Masaaki SAWA：“腺嘌呤连接的二-2-吡啶胺衍生物的金属配合物促进的 ATP 和 UTP 水解的反应性和位点选择性”Bull.Chem.Soc.Jpn.69 (3)。

K.Ogino, M.Sawa, and W.Tagaki: "Site-Selective Hydrolysis of ATP and UTP Promoted by Cu^<2+>-Complexes of Adenine-Linked 2,2'-Dipyridylamine Derivatives" The Fourth International Conference of Heteroatom Chemistry, Sheraton Walker Hill Hotel, Seoul, Korea

K.Ogino、M.Sawa 和 W.Tagaki：“Cu^<2>-腺嘌呤连接的 2,2-二吡啶胺衍生物的复合物促进 ATP 和 UTP 的位点选择性水解”第四届国际杂原子化学会议

Masaaki Sawa: "Reactivities and Site-Selectivities of Hydrolyses of ATP and UTP Promoted by Metal Complexes of Adenine-linked Di-2-pyridylamine Derivatives" Bull.Chem.Soc.Jpn.69(3月号掲載予定). (1996)

Masaaki Sawa：“Reactivities and Site-Selectivity of Hydrolyses of ATP and UTP Promotiond by Metal Complexes of Adenine-linked Di-2-pyridylamine Derivatives”Bull.Chem.Soc.Jpn.69（计划于 3 月号出版）（ 1996）