Study of functional model for metal containing hydrases by new tripot metal complexes

新型三罐金属配合物对含金属水合酶功能模型的研究

• 批准号: 12640535
• 负责人: FUJII Yuki
• 金额: $ 2.3万
• 依托单位: IBARAKI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12640535/
• 关键词: tripot; metal complex; hydrolysis; peptide; DNA; TACH; reaction mechanism; enzyme model; 銅(II); 亜鉛(II); 加水分解酵素; 切断; X線構造; speciation

Cu(II)TACH and Zn(II)TACH (TACH=cis, cis,1,3,5-triaminocyclohexane) were synthesized and their strained structures were confirmed by X-ray structure analysis. Hydrolysis and speciation study of several dipeptides with Cu(II)TACH was performed at pH 7-10, 70℃ to estimate the mechanism and the reaction intermediate. Hydrolysis of several depeptides with ZnCl_2, ZnHEN, ZnCD-AMP were followed to evaluate the acceleration effect. Also kinetic and thermodynamic data of free peptides (GlyGly, GlySer, SerGly, LeuGly, GlyLeu, SerMet, AlaSer, LeuSer, ValSer, GlyThr, AlaThr, GlyGlyGly) were measured at pH 7-10, 70℃. Hydrolytic cleavage of pBR322 DNA with Cu(II)TACH-X(X=H, Me, Et, 2Et, 3Et) were investigated at pH 7-9.5, 35℃ to study the reaction mechanism and the effect of X. Cyclic DNA(pHSG398, pUC19, pBR322, φX174, ColE1) were also cleaved with Cu(II)TACH to investigate the DNA size effect. Also, a new method to determine the scission site of DNA was established.

合成了铜(II)TACH和锌(II)TACH(TACH=顺式、顺式、1，3，5-三氨基环己烷)，其应变结构经X-射线结构分析确证。在pH为7~10、70℃的条件下，用铜(II)TACH对几种二肽进行了水解和形态研究，以推测其作用机理和反应中间体。用氯化锌、乙二醇锌、环腺苷锌对几种脱肽进行了水解实验，以评价其加速效果。在pH为7~10、70℃的条件下，测定了GlyGly、GlySer、Sergly、LeuGly、GlyLeu、Sermet、AlaSer、Leuser、Valser、GlyThr、AlaThr、GlyGlyGly的动力学和热力学数据。在pH 7~9.5，35℃的条件下，研究了铜(II)TACH-X(X=H，Me，Et，2Et，3Et)对pBR322DNA的切割作用，探讨了反应机理及X对环状φ的影响。此外，还建立了一种确定DNA断裂位置的新方法。

项目成果

Y.Fujii, et al.: "Metal-ion-assisted hydrolysis of dipeptides involving a serine residue in a neutral aqueous solution"Org.Biomol.Chem.. 1. 629-632 (2003)

Y.Fujii等人：“中性水溶液中涉及丝氨酸残基的二肽的金属离子辅助水解”Org.Biomol.Chem..1.629-632(2003)

F.Sakamoto, E.Suzuki, Y.Fujii: "Noel approach for the effective determination of DNA scission site using the Sanger method"J.Biochem.Biophys.Methods. 52. 97-109 (2002)

F.Sakamoto、E.Suzuki、Y.Fujii：“使用 Sanger 方法有效测定 DNA 断裂位点的 Noel 方法”J.Biochem.Biophys.Methods。

N.Yashiro, Y.Sonobe, A.Yamamura, T.Takarada, M.Komiyama, Y.Fujii: "Metal-ion-assisted hydrolysis of dipeptides involving a serine residue in a neutral aqueous solution"Org.Biomol.Chem.. 1. 629-632 (2003)

N.Yashiro、Y.Sonobe、A.Yamamura、T.Takarada、M.Komiyama、Y.Fujii：“中性水溶液中涉及丝氨酸残基的二肽的金属离子辅助水解”Org.Biomol.Chem..

T.Hirohama, M.Chikira, Y.Fujii: "EPR study of the dynamic behavior of cis, cis-1,3,5-triamino-cyclohexane Cu(II) complex on B-form DNA-fibers"J.Inourg.Biochem.. 98. 371-376 (2004)

T.Hirohama、M.Chikira、Y.Fujii：“顺式、顺式 1,3,5-三氨基-环己烷 Cu(II) 复合物在 B 型 DNA 纤维上动态行为的 EPR 研究”J.Inourg。

Y.Fujii, et al.: "Recent development of artificial nuclease with specificity of cleavage site"Recent Res.Devel.Nucleosides & Nucleotides. 2. 1-27 (2004)

Y.Fujii 等人：“具有切割位点特异性的人工核酸酶的最新开发”Recent Res.Devel.Nucleosides