荧光及ESI质谱法研究溶菌酶与磷酰化黄酮的相互作用

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. 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-Fen^1,2

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

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摘要/Abstract

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k_20℃=1.68×10⁴ L/mol,k_37℃=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 k_20℃=1.68×10⁴ L/mol and k_37℃=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

引用此文

陈晓岚, 于斐, 屈凌波, 赵玉芬. 荧光及ESI质谱法研究溶菌酶与磷酰化黄酮的相互作用[J]. 化学学报, 2004, 62(2): 188-193.

CHEN Xiao-Lan, YU Fei, QU Ling-Bo, ZHAO Yu-Fen. Study on the Interaction of Lysozyme and Diethyl Flavon-7-yl Phosphate by ESI-MS and Fluorescence Method[J]. Acta Chimica Sinica, 2004, 62(2): 188-193.

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[1] Liu, Z.-Z. Chinese Traditional and Herbal Drugs 1987, 18(1), 178 (in Chinese).(刘仲则, 中草药, 1987, 18(1), 178.)
[2] Wang, X.-L.; Xu, M.-X.; Xie, Y.-N. West China J. Pharm. Sci. 1999, 14(5-6), 309 (in Chinese).(王晓莉, 徐鸣夏, 谢益农, 华西药学杂志, 1999, 14(5-6), 309.)
[3] Chen, R.-Y.; Liu, L.-Z. Research on Organic Phosphorus, High Education Press, Beijing, 2002 (in Chinese)(陈茹玉, 刘纶祖著, 有机磷化学研究, 高等教育出版社, 北京, 2002.)
[4] Birendra, N. P.; Peter, L. B.; Urooj, A. M.; Liu, Y.-H.; Asshit, K. G. J. Mass Spectrom. 1998, 33, 911.
[5] Fenn, J. B.; Meng, C. K.; Wong, S. F.; Whitehouse, C. M. Science 1989, 246, 64.
[6] Chait, B. T.; Kent, S. B. H. Science 1992, 257, 1885.
[7] Smith, R. D.; Loo, J. A.; Edmonds, C. G.; Barinaga, C. J.; Udseth, H. R. Anal. Chem. 1990, 62, 882.
[8] Smith, D. L.; Zhang, Z. Mass Spectrom. Rev. 1994, 13, 411.
[9] Smith, R. D.; Light-Wahl, K. J. Biol. Mass Spectrom. 1993, 22, 493.
[10] Kelly, M. A.; Vesling, M. M.; Fenselaau, C. C.; Edmonds, C. G.; Smith, R. D. Rapid Commum. Mass Spectrom. 1991, 5, 101.
[11] He, M.-Y. Modern Organic and Biological Mass Spectrometry, Beijing University Press, Beijing, 2002 (in Chinese).(何美玉, 现代有机与生物质谱, 北京大学出版社, 北京, 2002.)
[12] Mann, M.; Meng, C.-K.; Fenn, J. B. Anal. Chem. 1989, 61, 1702.
[13] Verge, V.; Tristan, R.; Serge, M.; Suzanne, R. D.; Joseph-Claude, P.; Jean-Claude, M.; Jean-Pierre, M. Tetrahedron Lett. 2002, 43, 2363.
[14] Chen, G.-Z. The Methods of Fluoresence Analysis, Science Press, Beijing, 1975 (in Chinese).(陈国珍著, 荧光分析法, 科学出版社, 北京, 1975.)
[15] Lakowicz, J. R. Principle of Fluorescence Spectroscopy, Plenum Press, New York, 1983, pp. 257～295.
[16] Zhu, K.; Tong, S.-Y. Chem. J. Chin. Univ. 1996, 17(4), 53 (in Chinese).(朱铿, 童沈阳, 高等学校化学学报, 1996, 17(4), 53.)
[17] He, J.-X.; Jiang, C.-Q.; Wang, H.-J.; Wang, J.-Z. Chem. J. Chin. Univ. 1999, 20(10), 1548 (in Chinese).(贺吉香, 江崇球, 王洪鉴, 王敬政, 高等学校化学学报, 1999, 20(10), 1548.)
[18] Ma, G.-B.; Yang, P. Chin. J. Biochem. 1992, 8(5), 62 (in Chinese). (马贵斌, 杨频, 生物化学杂志, 1992, 8(5), 62.)
[19] Sarkar, B. Metal Ions in Biological Systems, Marcel Dekker, New York, 1991, p. 233.
[20] Yang, P. A Guide to Bioinorganic Chemistry, Xi′an Jiaotong University Press, Xi′an, 1991, p. 163 (in Chinese).(杨频著, 生物无机化学导论, 西安交通大学出版社, 西安, 1991, p. 163.)
[21] Yang, M.-M; Yang, P.; Zhang, L.-W. Chin. Sci. Bull. 1994, 39(1), 31 (in Chinese).(杨曼曼, 杨频, 张立伟, 科学通报, 1994, 39(1), 31.)

荧光及ESI质谱法研究溶菌酶与磷酰化黄酮的相互作用

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

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