多光谱法和分子对接模拟法研究黄腐酸与胃蛋白酶的相互作用

doi:10.7503/cjcu20190252

高等学校化学学报 ›› 2019, Vol. 40 ›› Issue (9): 1840.doi: 10.7503/cjcu20190252

多光谱法和分子对接模拟法研究黄腐酸与胃蛋白酶的相互作用

王晓霞^1,^*(),马力通¹,聂智华²,王正德¹,崔金龙¹,赵文渊¹,赛华征¹

^1. 内蒙古科技大学化学与化工学院, 包头 014010
^2. 清华大学生命科学学院, 北京 100084

收稿日期:2019-04-30 出版日期:2019-09-10 发布日期:2019-09-09
通讯作者: 王晓霞 E-mail:wxx572369@163.com
基金资助:
国家自然科学基金(21766025);内蒙古自治区科技计划项目(201702027);内蒙古自治区高等学校科学研究项目(No.NJZY17166)

Interaction Between Fulvic Acid and Pepsin Investigated by Multispectral and Molecular Docking Simulation ^†

WANG Xiaoxia^1,^*(),MA Litong¹,NIE Zhihua²,WANG Zhengde¹,CUI Jinlong¹,ZHAO Wenyuan¹,SAI Huazheng¹

^1. School of Chemistry and Chemical Engineering, Inner Mongolia University of Science and Technology,Baotou 014010, China
^2. School of Life Sciences, Tsinghua University, Beijing 100084, China

Received:2019-04-30 Online:2019-09-10 Published:2019-09-09
Contact: WANG Xiaoxia E-mail:wxx572369@163.com
Supported by:
? Supported by the National Natural Science Foundation of China(21766025);the Inner Mongolia Autonomous Region Science and Technology Plan Funding Project, China(201702027);the Inner Mongolia Autonomous Region Higher Education Research Project, China(No.NJZY17166)

摘要/Abstract

摘要：

利用荧光光谱、同步荧光光谱、三维荧光光谱、紫外-可见吸收光谱、傅里叶变换红外光谱和圆二色光谱以及分子对接模拟方法研究了黄腐酸(FA)与胃蛋白酶(PEP)之间的相互作用. 荧光光谱分析表明, FA-PEP荧光猝灭的类型为静态猝灭. 根据Stern-Volmer方程和静态猝灭双对数公式计算得到猝灭常数K_sv和结合位点数n. 根据Vant’t Hoff方程计算得到热力学常数ΔH=-59.86 kJ/mol, ΔS=-98.13 J·mol ^-1·K ^-1, ΔG=-30.62 kJ/mol(298 K). 热力学分析表明, 氢键和范德华力是PEP与FA之间的主要结合力, 其反应为自发过程. 根据F?rster非辐射能量转移理论, 计算得到PEP和FA之间的结合距离为2.436 nm, 表明在FA与PEP之间发生了非辐射能量转移. 三维荧光光谱分析表明, 在FA存在下PEP的肽链骨架结构发生了改变. 此外, 紫外-可见吸收光谱、同步荧光光谱和红外光谱结果表明, FA使PEP的二级构象发生变化. 分子对接模拟结果表明, FA引起PEP荧光猝灭的结合作用力不仅有氢键和范德华力, 还有疏水作用力.

关键词: 黄腐酸, 胃蛋白酶, 多光谱法, 分子对接模拟法

Abstract:

The interaction of fulvic acid(FA) with pepsin(PEP) was investigated using fluorescence spectra, UV-Vis absorption spectra, three-dimensional fluorescence spectra, synchronous fluorescence spectra, Fourier transform infrared spectroscopy, circular dichroism and molecular docking simulation method. Fluorescence spectrum analyses shows that the type of fluorescence quenching by FA-PEP is static quenching. According to the Stern-Volmer equation and the static quenching double logarithmic formula, the quenching constants(K_sv) and the number of binding sites(n) were calculated. According to the Vant’t Hoff equation, the thermodynamic parameters were calculated to be ΔH=-59.86 kJ/mol, ΔS=-98.13 J·mol ^-1·K ^-1 and ΔG=-30.62 kJ/mol(298 K). Thermodynamic analysis suggests that hydrogen bonds and van der Waal’s forces are the main forces between PEP and FA, and the interaction is spontaneous process. According to the theory of F?rster’s non-radiative energy transfer, the binding distance between PEP and FA was calculated to be 2.436 nm, implying that non-radiative energy transfer occurs between FA. The molecular docking simulation technique displays that the binding force of FA-PEP includes not only hydrogen bonds and van der Waals forces but also hydrophobic interaction forces.

Key words: Fulvic acid, Pepsin, Multispectroscopy, Molecular docking simulation method

中图分类号:

O657.3

TrendMD:

王晓霞, 马力通, 聂智华, 王正德, 崔金龙, 赵文渊, 赛华征. 多光谱法和分子对接模拟法研究黄腐酸与胃蛋白酶的相互作用. 高等学校化学学报, 2019, 40(9): 1840.

WANG Xiaoxia, MA Litong, NIE Zhihua, WANG Zhengde, CUI Jinlong, ZHAO Wenyuan, SAI Huazheng. Interaction Between Fulvic Acid and Pepsin Investigated by Multispectral and Molecular Docking Simulation ^†. Chem. J. Chinese Universities, 2019, 40(9): 1840.

图/表 10

Fig.1 Fluorescence quenching spectra of FA-PEP interaction system at 298 K

Table 1 Stern-Volmer linear equations correlation coefficients and thermodynamic parameters

T/K	10^-4K_sv/ (L·mol^-1)	10^-12K_q/ (L·mol^-1·s^-1)	10^-5K_A/ (L·mol^-1)	Binding-site number, n	ΔH/ (kJ·mol^-1)	ΔS/ (J·mol^-1·K^-1)	ΔG/ (kJ·mol^-1)
298	9.906	9.906	2.421	1.368			-30.62
303	9.089	9.089	1.435	1.322	-59.86	-98.13	-30.12
308	8.860	8.860	1.107	1.294			-29.63

Fig.2 Overlapping of the fluorescence spectrum of PEP(a) with the absorption spectrum of FA(b) a. c(PEP)=5.0×10-5 mol/L; b. c(FA)=5.0×10-5 mol/L. T=298 K, pH=2.0.

Fig.3 Synchronous fluorescence of FA and PEP (1)Δλ=15 nm; (B) Δλ=60 nm. c(PEP)=5.0×10-5 mol/L. 104c(FA)/(mol·L-1): a—i. 0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6. T=298 K, pH=2.0.

Fig.4 UV spectra of FA and PEP c(PEP)=6.0×10-4 mol/L; 104c(FA)/(mol·L-1): a—i. 0, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4, 1.6. T=298 K, pH=2.0.

Fig.5 Three-dimensional fluorescence spectra(A) and contour diagrams(B) of PEP c(PEP)=5×10-5 mol/L, T=298 K, pH=2.0.

Fig.6 Three-dimensional fluorescence spectra(A) and contour diagrams(B) of FA-PEP system c(PEP)=5×10-5 mol/L, c(FA)=5×10-5 mol/L, T=298 K, pH=2.0.

Fig.7 Circular dichroism spectra of the PEP-FA system a. c(PEP)=5.0×10-5 mol/L, c(PEP)∶c(FA)=1∶0; b. c(PEP)=5.0×10-5 mol/L, c(FA)=2.5×10-4 mol/L, c(PEP)∶c(FA)=1∶5. T=298 K, pH=2.0.

Fig.8 Infrared spectra of PEP(a) and FA-PEP system[m(PEP)∶m(FA)=1∶1](b) at 298 K

Fig.9 Molecular docking simulation of FA and PEP

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多光谱法和分子对接模拟法研究黄腐酸与胃蛋白酶的相互作用

Interaction Between Fulvic Acid and Pepsin Investigated by Multispectral and Molecular Docking Simulation ^†

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多光谱法和分子对接模拟法研究黄腐酸与胃蛋白酶的相互作用

Interaction Between Fulvic Acid and Pepsin Investigated by Multispectral and Molecular Docking Simulation †

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Interaction Between Fulvic Acid and Pepsin Investigated by Multispectral and Molecular Docking Simulation ^†