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同步荧光光谱探究头孢西丁钠与溶菌酶的结合机制

张红彩,刘保生,程旭

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张红彩, 刘保生, 程旭. 同步荧光光谱探究头孢西丁钠与溶菌酶的结合机制[J]. , 2020, 13(3): 492-500. doi: 10.3788/CO.2019-0112
引用本文: 张红彩, 刘保生, 程旭. 同步荧光光谱探究头孢西丁钠与溶菌酶的结合机制[J]. , 2020, 13(3): 492-500.doi:10.3788/CO.2019-0112
ZHANG Hong-cai, LIU Bao-sheng, CHENG Xu. Study on the binding mechanism of cefoxitin sodium to lysozyme by synchronous fluorescence spectroscopy[J]. Chinese Optics, 2020, 13(3): 492-500. doi: 10.3788/CO.2019-0112
Citation: ZHANG Hong-cai, LIU Bao-sheng, CHENG Xu. Study on the binding mechanism of cefoxitin sodium to lysozyme by synchronous fluorescence spectroscopy[J].Chinese Optics, 2020, 13(3): 492-500.doi:10.3788/CO.2019-0112

同步荧光光谱探究头孢西丁钠与溶菌酶的结合机制

doi:10.3788/CO.2019-0112
基金项目:国家自然科学基金资助项目(No.21375032)
详细信息
    作者简介:

    张红彩(1993—),女,河北石家庄人,硕士研究生,2017年于保定学院获得学士学位,主要从事分子发光学理论与应用研究。E-mail:1179497976@qq.com

    刘保生(1963—),男,河北保定人,硕士,研究员,1986年、1992年于河北大学分别获得分析化学专业学士、硕士学位,主要从事分子发光学理论与应用研究。E-mail:lbs@hbu.edu.cn

    通讯作者:

    刘保生(1963—),男,河北保定人,硕士,研究员,1986年、1992年于河北大学分别获得分析化学专业学士、硕士学位,主要从事分子发光学理论与应用研究。E-mail:lbs@hbu.edu.cn

  • 中图分类号:O657.3

Study on the binding mechanism of cefoxitin sodium to lysozyme by synchronous fluorescence spectroscopy

Funds:Supported by National Natural Science Foundation of China (No. 21375032)
More Information
  • 摘要:在模拟生理学条件下(pH=7.40),采用同步荧光法研究了头孢西丁钠(CFXS)和溶菌酶(LYSO)中的荧光基团酪氨酸(Tyr)残基、色氨酸(Trp)残基之间的相互作用。结果表明:CFXS以静态猝灭的方式猝灭LYSO中的Tyr、Trp残基的荧光,结合位点数 n≈1。310 K时,Tyr与Trp残基反应的荧光猝灭比率分数 N SFQR(Trp)(60.25%)> N SFQR(Tyr)(39.75%),结合位置更靠近Trp残基。Hill系数 n H约为1,表明CFXS与LYSO中Tyr与Trp残基的结合不会影响后继配体与蛋白质的结合。CFXS与LYSO中Tyr残基的药物结合率 W( Q)为0.19%~0.13%,Trp残基的药物结合率 W( Q)为0.23%~0.14%,游离的药物含量几乎不变,这表明CFXS与LYSO中Tyr与Trp残基的结合基本不影响药物的疗效。Tyr残基的蛋白结合率 W( B)为52.69%~54.67%,Trp残基的蛋白结合率 W( B)为67.67%~69.39%,因此,蛋白中游离的氨基酸残基数目会明显降低。CFXS-LYSO结合体系的主要作用力类型是疏水作用,分子对接结果表明CFXS与LYSO之间还存在氢键作用,且两者的最佳结合位置在LYSO的活性中心附近,两者的结合改变了活性中心处氨基酸残基的微环境。

  • 图 1CFXS-LYSO体系的同步荧光光谱 (T=310 K) (a) Δλ=15 nm;(b) Δλ=60 nm

    Figure 1.Synchronous fluorescence spectra of CFXS-LYSO system (T=310 K) (a) Δλ=15 nm; (b) Δλ=60 nm

    图 2CFXS-LYSO体系荧光强度随NaCl浓度的变化 (T=310 K)

    Figure 2.Fluorescence intensity of CFXS-LYSO system as a function of NaCl concentrationCNacl(T=310 K)

    图 3CFXS与LYSO相互作用的分子对接图

    Figure 3.Molecular docking model of the interaction between CFXS and LYSO

    表 1CFXS-LYSO体系的同步荧光猝灭反应参数

    Table 1.Reactive parameters of synchronous fluorescence quenching for CFXS-LYSO system

    Δλ/nm T/K Ksv/L·mol−1·s−1 kq/L·mol−1 r1 Ka/L·mol−1 n r2
    Δλ=60 298 2.18×104 2.18×1012 0.990 5 1.73×104 1.18 0.992 2
    310 1.61×104 1.61×1012 0.992 4 1.58×104 1.03 0.991 3
    318 1.26×104 1.26×1012 0.994 1 1.39×104 0.96 0.993 5
    Δλ=15 298 1.33×104 1.33×1012 0.993 8 0.96×104 1.07 0.995 7
    310 1.17×104 1.17×1012 0.991 4 0.84×104 1.17 0.992 2
    318 0.83×104 0.83×1012 0.995 2 0.68×104 1.12 0.993 4
    r1为方程I0/I~[L]的线性相关系数;r2为方程lg[(I0-I)/I]~lg{[L]-n[Bt](I0-I)/I0}的线性相关系数;[Bt]=5.0×10−7mol/L。
    下载: 导出CSV

    表 2不同温度下CFXS-LYSO体系的热力学参数

    Table 2.Thermodynamic parameters of CFXS-LYSO system at different temperatures

    System T/K Ka/L·mol−1 ΔH/kJ·mol−1 ΔS/J·mol−1K−1 ΔG/kJ·mol−1
    Δλ=15 nm 298 0.96×104 −13.15 32.10 −22.71
    310 0.84×104 32.70 −23.28
    318 0.68×104 32.01 −23.33
    Δλ=60 nm 298 1.73×104 −11.68 41.93 −24.17
    310 1.58×104 42.70 −24.91
    318 1.39×104 42.58 −25.22
    下载: 导出CSV

    表 3不同温度下CFXS和LYSO中Tyr、Trp残基的Hill系数

    Table 3.Hill coefficients of Tyr and Trp residues in CFXS and LYSO at different temperatures

    T/K Δλ=60 nm Δλ=15 nm
    nH r3 nH r3
    298 0.94 0.993 7 0.99 0.997 1
    310 1.15 0.994 8 1.07 0.993 3
    318 0.92 0.996 7 0.94 0.995 2
    nH为体系的Hill系数;r3为方程lg[Y/(1-Y)]~lg[L]的线性相关系数。
    下载: 导出CSV

    表 4CFXS-LYSO体系的对接能量(单位:kJ/mol)

    Table 4.Docking energy of CFXS-LYSO system (unit: kJ/mol)

    Protein PDB ID ΔG0 ΔE1 ΔE2 ΔE3
    2LYZ −24.81 −39.78 −36.44 −3.34
    下载: 导出CSV
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  • 收稿日期:2019-07-23
  • 修回日期:2019-08-20
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