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化学学报
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化学学报 ›› 2004, Vol. 62 ›› Issue (2): 188-193. 上一篇    下一篇

研究论文

荧光及ESI质谱法研究溶菌酶与磷酰化黄酮的相互作用

陈晓岚1, 于斐1, 屈凌波1, 赵玉芬1,2   

  1. 1. 郑州大学化学系, 化学生物学重点实验室, 郑州, 450052;
    2. 清华大学教育部生命有机磷化学和化学生物重点实验室, 北京, 100084
  • 投稿日期:2003-05-08 修回日期:2003-09-24 发布日期:2014-01-26
  • 通讯作者: 屈凌波,E-mail:qulingbo@zzu.edu.cn;Fax:0371-7763220 E-mail:qulingbo@zzu.edu.cn

Study on the Interaction of Lysozyme and Diethyl Flavon-7-yl Phosphate by ESI-MS and Fluorescence Method

CHEN Xiao-Lan1, YU Fei1, QU Ling-Bo1, ZHAO Yu-Fen1,2   

  1. 1. Department of Chemistry, Key Laboratory of Chemistry and Biology, Zhengzhou University, Zhengzhou 450052;
    2. Key Laboratory for Bioorganic Phosphorus Chemistry, Ministry of Education, Department of Chemistry, School of Life Sciences and Engineering, T
  • Received:2003-05-08 Revised:2003-09-24 Published:2014-01-26

PDF

410

被引次数

69

分别用荧光法和ESI质谱法研究了磷酰化黄酮和溶菌酶的相互作用.结果均显示磷酰化黄酮能够和溶菌酶发生弱相互作用,与黄酮相比它对溶菌酶更具亲和力.根据荧光猝灭双倒数图计算了磷酰化黄酮与溶菌酶之间的结合常数为k20℃=1.68×104 L/mol,k37℃=1.06×104 L/mol,实验证明随着温度的升高,磷酰化黄酮与溶菌酶的结合常数逐渐降低,说明了两者之间形成了复合物,此荧光猝灭过程为静态猝灭.根据Föster能量传递原理计算出磷酰化黄酮在溶菌酶上的结合距离,并根据热力学参数确定了磷酰化黄酮与溶菌酶之间的作用力类型为电荷作用力.

关键词: ESI, 荧光猝灭, 溶菌酶, 磷酰化黄酮, 能量传递

ESI MS and fluorescence methods were used to study the interactions between lysozyme and diethyl flavon 7-yl phosphate and 7-hydroxyflavone. The results showed that the phosphorylated flavonoid could form non covalent complexes lysozyme and showed higher binding affinity with the protein than 7-hydroxyflavone did. The association constants of lysozyme and diethyl flavon 7-yl phosphate were determined from a double reciprocal Lineweaver Burk plot. The binding constants were k20℃=1.68×104 L/mol and k37℃=1.06×104 L/mol respectively. Experiments demonstrated that the higher the temperature was, the lower the slop of quenching curve of lysozyme was in presence of different amounts of diethyl flavon 7-yl phosphate. It was confirmed that the combination of diethyl flavon 7-yl phosphate with lysozyme was a single static quenching process. According to the Föster dipole dipole energy transfer, the distance between the diethyl flavon 7-yl phosphate and tryptophane was measured. From thermodynamical coordination it could be judged that the binding power between diethyl flavon 7-yl phosphate and lysozyme was static electric power.

Key words: ESI, fluorescence quenching, lysozyme, diethyl flavon-7-yl phosphate, energy transition