双核钌配合物与c-myc G-四链体DNA的键合性能

doi:10.3969/j.issn.1000-1565.2021.01.004

摘要/Abstract

摘要： 通过紫外可见吸收光谱和荧光光谱滴定、Job plot、圆二色光谱、聚合酶链式扩增反应(PCR)、显色反应以及FRET实验,研究了苯酚基双核钌配合物与c-myc G-四链体DNA(c-myc Pu27和c-myc Pu22)之间的相互作用. 结果表明,配合物对c-myc Pu27和c-myc Pu22 DNA都有较强结合,键合常数分别为6.21 × 10⁷ L/mol 和2.33×10⁶ L/mol,键和模式为沟槽结合,键合物质的量比为2∶1;配合物在浓度为4 μmol/L(c-myc Pu27)、6 μmol/L(c-myc Pu22)时,可完全抑制PCR扩增产物生成,表明其能够诱导c-myc DNA形成G-四链体结构. 此外,配合物可使F27T和F22T的熔解温度分别升高13 ℃和12 ℃,且在双链DNA存在时F27T和F22T的熔解温度几乎没有变化,表明其对c-myc G-四链体DNA具有显著的稳定性及选择性.

关键词: 钌配合物, 四链体, DNA

Abstract: The interactions of a binuclear phenol-containing ruthenium(Ⅱ)complex with c-myc G-quadruplex DNA(c-myc Pu27 and c-myc Pu22)were investigated by UV-visible absorption spectra and fluorescence spectra titrations, Job plot, CD spectra, PCR(polymerase chain reaction)stop assay, color reaction, and FRET(fluorescence resonance energy transfer)experiments. The complex bind to c-myc Pu27 and c-myc Pu22 DNA strongly with the bonding constants of 6.21 × 10⁷ L/mol(c-myc Pu27)and 2.33×10⁶ L/mol(c-myc Pu22)via groove mode and the binding stoichiometry is 2∶1. The replication of c-myc DNA was completely blocked by the complex at concentrations of 4 μmol/L for c-myc Pu27 and 6 μmol/L for c-myc Pu22, indicating that the complex can induce c-myc DNA to form a G-quadruplex structure. In addition, the complex can increase the melting temperatures of F27T and F22T by 13 ℃(c-myc Pu27)and- DOI:10.3969/j.issn.1000-1565.2021.01.004双核钌配合物与c-myc G-四链体DNA的键合性能严双美¹,米雅萱¹,崔玥¹,郑泽宝²,赵晓珑¹ (1. 河北大学化学与环境科学学院, 河北保定 071002;2. 泰山学院化学化工学院,山东泰安 271021)摘要:通过紫外可见吸收光谱和荧光光谱滴定、Job plot、圆二色光谱、聚合酶链式扩增反应(PCR)、显色反应以及FRET实验,研究了苯酚基双核钌配合物与c-myc G-四链体DNA(c-myc Pu27和c-myc Pu22)之间的相互作用. 结果表明,配合物对c-myc Pu27和c-myc Pu22 DNA都有较强结合,键合常数分别为6.21 × 10⁷ L/mol 和2.33×10⁶ L/mol,键和模式为沟槽结合,键合物质的量比为2∶1;配合物在浓度为4 μmol/L(c-myc Pu27)、6 μmol/L(c-myc Pu22)时,可完全抑制PCR扩增产物生成,表明其能够诱导c-myc DNA形成G-四链体结构. 此外,配合物可使F27T和F22T的熔解温度分别升高13 ℃和12 ℃,且在双链DNA存在时F27T和F22T的熔解温度几乎没有变化,表明其对c-myc G-四链体DNA具有显著的稳定性及选择性.关键词:钌配合物;四链体;DNA中图分类号:Q523 文献标志码:A 文章编号:1000-1565(2021)01-0023-09Binding properties of a dinuclear ruthenium complex to c-myc G-quadruplex DNAYAN Shuangmei¹, MI Yaxuan¹, CUI Yue¹, ZHENG Zebao², ZHAO Xiaolong¹(1. College of Chemistry and Environmental Science, Hebei University, Baoding 071002, China; 2. College of Chemistry and Chemical Engineering, Taishan University, Taian 271021, China)Abstract: The interactions of a binuclear phenol-containing ruthenium(Ⅱ)complex with c-myc G-quadruplex DNA(c-myc Pu27 and c-myc Pu22)were investigated by UV-visible absorption spectra and fluorescence spectra titrations, Job plot, CD spectra, PCR(polymerase chain reaction)stop assay, color reaction, and FRET(fluorescence resonance energy transfer)experiments. The complex bind to c-myc Pu27 and c-myc Pu22 DNA strongly with the bonding constants of 6.21 × 10⁷ L/mol(c-myc Pu27)and 2.33×10⁶ L/mol(c-myc Pu22)via groove mode and the binding stoichiometry is 2∶1. The replication of c-myc DNA was completely blocked by the complex at concentrations of 4 μmol/L for c-myc Pu27 and 6 μmol/L for c-myc Pu22, indicating that the complex can induce c-myc DNA to form a G-quadruplex structure. In addition, the complex can increase the melting temperatures of F27T and F22T by 13 ℃(c-myc Pu27)and- 收稿日期:2020-04-20 基金项目:国家自然科学基金资助项目(21401038);河北省自然科学基金资助项目(B2016201122);保定市科学技术研究与发展计划项目(14ZF009);泰安市科学技术研究与发展计划项目(2015GX2048) 第一作者:严双美(1995—),女,河北秦皇岛人,河北大学在读硕士研究生.E-mail:2986558284@qq.com 通信作者:赵晓珑(1980—),女,河北邯郸人,河北大学副教授,博士,主要从事钌配合物与生物大分子相互作用研究.E-mail: longlong_666@sina.com郑泽宝(1980—),男,山东泰安人,泰山学院副教授,博士,主要从事钌配合物生物传感研究.E-mail: zhengzebao@163.com第1期严双美等:双核钌配合物与c-myc G-四链体DNA的键合性能12 ℃(c-myc Pu22), and the melting temperatures of F27T and F22T were almost unchanged in the presence of double-stranded DNA, suggesting that the complex has significant stability and selectivity for c-myc G-quadruplex DNA.

Key words: ruthenium complex, quadruplex, DNA

中图分类号:

Q523

严双美,米雅萱,崔玥,郑泽宝,赵晓珑. 双核钌配合物与c-myc G-四链体DNA的键合性能[J]. 河北大学学报(自然科学版), 2021, 41(1): 23-31.

YAN Shuangmei, MI Yaxuan, CUI Yue, ZHENG Zebao, ZHAO Xiaolong. Binding properties of a dinuclear ruthenium complex to c-myc G-quadruplex DNA[J]. Journal of Hebei University(Natural Science Edition), 2021, 41(1): 23-31.

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