高等学校化学学报 ›› 2017, Vol. 38 ›› Issue (8): 1341-1346.doi: 10.7503/cjcu20170103

• 分析化学 • 上一篇    下一篇

离子淌度质谱结合荧光光谱法研究5-氟尿苷与铜锌超氧化物歧化酶的相互作用

赵兵1,2, 庄小禹1, 刘舒1, 郑重1, 刘志强1, 宋凤瑞1   

  1. 1. 中国科学院长春应用化学研究所国家质谱中心(长春)&吉林省中药化学与质谱重点实验室, 长春 130022;
    2. 中国科学技术大学, 合肥 230029
  • 收稿日期:2017-02-22 修回日期:2017-07-12 出版日期:2017-08-10 发布日期:2017-07-12
  • 通讯作者: 宋凤瑞,女,博士,研究员,博士生导师,主要从事质谱方法、代谢及代谢组学研究.E-mail:songfr@ciac.ac.cn E-mail:songfr@ciac.ac.cn
  • 基金资助:

    国家自然科学基金(批准号:21673219,81530094)资助.

Interaction Between 5-Furd and Cu,Zn-SOD1 by Ion Mobility Mass Spectrometry and Fluorescence Spectroscopy

ZHAO Bing1,2, ZHUANG Xiaoyu1, LIU Shu1, ZHENG Zhong1, LIU Zhiqiang1, SONG Fengrui1   

  1. 1. National Center of Mass Spectrometry in Changchun & Jilin Province Key Laboratory of Chinese Medicine Chemistry and Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China;
    2. University of Science and Technology of China, Hefei 230029, China
  • Received:2017-02-22 Revised:2017-07-12 Online:2017-08-10 Published:2017-07-12
  • Supported by:

    Supported by the National Natural Science Foundation of China(Nos.21673219, 81530094).

摘要:

分别利用离子淌度质谱和荧光光谱方法研究了5-氟尿苷(5-Furd)与铜锌超氧化物歧化酶(SOD1)的相互作用.离子淌度质谱研究结果表明,5-Furd能够稳定SOD1构型,减缓其碰撞诱导去折叠;外源荧光光谱方法进一步证明5-Furd可以抑制SOD1的聚集.利用荧光光谱研究了不同温度下5-Furd与SOD1的相互作用,阐明了二者的作用机制、结合常数和结合位点数,并根据热力学函数推断二者的作用力类型为典型的疏水作用.

关键词: 电喷雾电离-离子淌度质谱, 荧光光谱, 铜锌超氧化物歧化酶, 5-氟尿苷, 相互作用

Abstract:

The interaction between 5-Furd and Cu, Zn superoxide dismutase(SOD1) was investigated by means of electrospray-ion mobility mass spectrometry(ESI-IMS-MS) and fluorescence spectroscopy. The result of ESI-IMS-MS showed that 5-Furdis could stabilize the conformation of SOD1 against unfolding transition at different collision voltages. Through the ThT fluorescence method, it was further testified that 5-Furd could inhibit the aggregation of apo-SOD1. In addition, the interaction between SOD1 and 5-Furd at different temperatures was investigated by fluorescence spectrometry, and the characters of their interaction were elucidated including the interaction mechanism, the binding constant, binding site number and the corresponding thermo-dynamic parameters(△H, △S and △G). According to the thermo-dynamic parameters of interaction, it was found that the hydrophobic forces play an important role on their interaction.

Key words: Electrospray-ion mobility mass spectrometry(ESI-IMS-MS), Fluorescence spectroscopy, Cu, Zn superoxide dismutase(SOD1), 5-Furd, Interaction

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