Development of zinc detection system with novel zinc-fluorophores

新型锌荧光团锌检测系统的开发

• 批准号: 12559006
• 负责人: KOIKE Tohru
• 金额: $ 5.06万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12559006/
• 关键词: Zinc Fuorophore; Zinc(II) Ion; Fluorometry; Macrocyclic Polyamine; Chelate; 大環状ポリアミン

Quantitative analysis of trace zinc(II) ion with a selective analytical reagent has become extremely important for environmental and biological applications. While remarkable development has been made for other biologically important divalent metal ions, in particular calcium(II) ion, with several selective fluorophores such as Quin-2 and Fura-2, there have been few zinc(II)-selective analytical reagents available for biological research on zinc(II) ion. Currently, the most widely used probes for detecting intracellular zinc(II) ion are 8-aminoquinoline sulfonamide derivatives such as Zinquine. Recently, fluorescein-appended zinc(II) chelates such as azacrownfluorone have been developed as improved proves for understanding the zinc(II) chemistry in biological systems. While working on elucidation of intrinsic properties of zinc(II) ion in the catalytic sites of zinc enzymes by use of macrocyclic polyamine complexes (e.g., zinc(II) complexes with 1,5,9-triazacyclododecane ([12]aneN_3) a … More nd 1,4,7,10-tetraazacyclododecane (cyclen)), we have discovered unique acid properties of zinc(II) ion. One of the most outstanding zinc(II) properties is a strong affinity to aromatic sulfonamides, as illustrated by formation of the strong bonds between the deprotonated sulfonamide N^- anions and zinc(II) in the macrocyclic polyamines at physiological pH. On the basis of these findings, we have designed 5-dimethylamino-naphthalene-1-sulfonic acid [2-(1,4,7,10-tetraazacyclododec-1-yl)-ethyl]-amide (dansylamidoethyl-cyclen) that serves as a novel type of zinc(II)-fluorophore by forming a highly fluorescent 1:1 complex with zinc(II) ion at physiological pH. The zinc(II) dissociation constant is much smaller than intracellular zinc(II) concentrations, which is a drawback on the dynamic analysis of intracellular zinc(II) ion. In order to shift the analytical range to the higher concentrations (I.e., biological conditions), we have designed some zinc-fluorophores (e.g., dansylamidoethyl-[12]aneN_3) having a larger K_d value. In this study we deterimined the synthetic method and characterization of those novel fluorophores and those zinc(II) complexes. Less

具有选择性分析试剂的痕量锌离子（II）离子的定量分析对于环境和生物学应用非常重要。虽然已经为其他重要的生物学二价金属离子（尤其是钙（II）离子进行了显着的发展，具有几种选择性的荧光团，例如Quin-2和Fura-2，但很少有锌（II）选择性分析试剂可用于锌（II）离子的生物学研究。目前，检测细胞内锌（II）离子的最广泛使用的问题是8-氨基喹啉磺酰胺衍生物，例如Zinquine。最近，荧光素成熟的锌（II）螯合物（如Azacrownfluorone）已被开发，因为改进的证明是了解生物系统中锌（II）化学的证明。通过使用大环多胺复合物（例如，具有1,5,5,9- triazacyclododecane（[12] Anen_3）A…More ND 1,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,4,l，在锌酶催化位点阐明锌（II）离子的固有性能的同时（II）。锌离子的独特酸性能。最杰出的锌（II）性质之一是与芳族磺酰胺的强亲和力，如生理pH值的大环聚胺中的去质子甲酰胺N^ - 阴离子和锌（II）之间的强键所示。根据这些发现，我们设计了5-二甲基氨基氨基1-磺酸[2-（1,4,7,10-二甲基二硫酸） - 抗氧基] -Amide（Dansylamidoethylamidoethyal-乙基） - 与Zinc（II） - 富集的ZINC（II）相关（II）（II）（II）（II）（II）（II）（II）（II）（II）（II）（II）（II）（II）（II）（1） ph。锌（II）解离常数比细胞内锌（II）浓度小得多，这是对细胞内锌（II）离子动态分析的缺点。为了将分析范围转移到较高的浓度（即生物条件），我们设计了一些具有较大K_D值的锌氟噬菌体（例如Dansylamidoethyl- [12] anen_3）。在这项研究中，我们确定了那些新型荧光团和那些锌（II）复合物的合成方法和表征。较少的

项目成果

Emiko Kinoshita-Kikuta: "A novel procedure for simple and efficient genotyping of single nucleotide polymorphisms by using the Zn-cyclen complex"Nucleic Acid Research. 30. e126(1-6) (2002)

Emiko Kinoshita-Kikuta：“利用 Zn-cyclen 复合物对单核苷酸多态性进行简单有效的基因分型的新方法”核酸研究。

Eiichi Kimura: "Elaboration of Selective and Efficient Recognition of Tymine Base in Dinucleotides, Single-stranded and Double-stranded DNA by Zn(II)-acrydinylcyclen"J. Am. Chem. Soc.. 122. 4668-4677 (2000)

Eiichi Kimura：“Zn(II)-acrydinylcyclen 对二核苷酸、单链和双链 DNA 中酪氨酸碱基的选择性和高效识别的阐述”J。

Tohru Koike: "Synthesis and characterization of the zinc(II)-fluorophore, 5-dimethylamino-naphthalene-1-sulfonic acid [2-(1, 5, 9-triazacyclo-dodec-1-yl)-ethyl]-amide and its zinc(II) complex"J.Chem.Soc., Dalton Transactions. (in press). (2002)

Tohru Koike：“锌(II)-荧光团、5-二甲氨基-萘-1-磺酸[2-(1, 5, 9-三氮杂环十二-1-基)-乙基]-酰胺的合成和表征以及

Emiko Kikuta: "Controlling gene expression by zinc(II)-macrocyclic tetraamine complexes"Journal of Inorganic Biochemistry. 79. 253-259 (2001)

Emiko Kikuta：“通过锌(II)-大环四胺复合物控制基因表达”无机生物化学杂志。

Eiichi Kimura: "Elaboration of Selective and A Efficient Recognition of Thymine Base in Dinucleotides, Single-stranded, and Double-stranded DNA by Zn-Acridinycyclen"J.Am.Chem.Soc.. 122. 4668-4677 (2000)

Eiichi Kimura：“通过 Zn-Acridinycyclen 对二核苷酸、单链和双链 DNA 中的胸腺嘧啶碱基进行选择性和有效识别的阐述”J.Am.Chem.Soc.. 122. 4668-4677 (2000)