Interaction between Fleroxacin and Lysozyme by Using Multi-Spectral Techniques and Molecular Docking
FANG Qing1, WANG Yi-run1, GUO Chen-hui1, LIU Ying1,2*
1. College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China
2. Beijing Engineering Research Center of Food Environment and Public Health, Minzu University of China, Beijing 100081, China
Abstract:The interaction of fleroxacin (FLE) with extracelluar protein (lysozyme, LYSO) was investigated by using multi-spectral techniques and molecular docking in vitro. Fluorescence spectra studies indicated that FLE quenched LYSO fluorescence in a static mode with binding constants (Ka) of 4.10×104 and 0.74×104 L·mol-1 at 298 and 310 K, respectively. The thermodynamic parameters demonstrated that hydrogen bonds and van der Waals forces played the major role in the binding process. Based on the Frster theory of nonradiative energy transfer, the binding distance (r) between FLE and the inner tryptophan residues of LYSO was calculated to be 3.16 nm. The Hill’s coefficient (nH) was calculated by using involved equations, implying the negative cooperativity between them. Circular dichroism spectra (CD) indicated the secondary structure of LYSO was partially destroyed by FLE with the α-helix percentage decreasing from 21.1% to 8.8%. UV-Vis spectral, synchronous fluorescence and three-dimensional fluorescence spectra revealed the binding interaction could cause conformational and micro-environmental changes of LYSO. The results of molecular docking showed that FLE was mainly bound in the active site hinge region where ASP-52, TRP-62 and TRP-63 were located, and well supported the thermodynamic results. Besides, FIE made the activity of lysozyme decrease with the increasing concentration of fleroxacin, indicating that structural changes in lysozyme can cause the inhibition of lysozyme activity. The work clarified the interaction mechanism of FLE with LYSO at molecular level.
基金资助: The National Natural Science Foundation of China (21177163), 111 Project B08044, Special Guidance Fund of Building World First-class Universities (Disciplines) and Characteristic Development of Minzu University of China (2016), Coordinate Development of First-Class and First-Class University Discipline Construction Funds(10301-0150200604), The Academic Team Construction Project of Minzu University of China (2015MDTD25C&13C), First-class Universities and First-class Discipline Construction Transitional Funds Under Special Funding (2016, Master)
通讯作者:
刘 颖
E-mail: liuying4300@163.com
作者简介: FANG Qing, (1992—), master of College of Life and Environmental Sciences, Minzu University of China
e-mail:
fq19920119@126.com
引用本文:
方 庆,王艺润,郭晨辉,刘 颖. 光谱法和分子对接技术研究氟罗沙星与溶菌酶的相互作用[J]. 光谱学与光谱分析, 2018, 38(02): 654-659.
FANG Qing, WANG Yi-run, GUO Chen-hui, LIU Ying. Interaction between Fleroxacin and Lysozyme by Using Multi-Spectral Techniques and Molecular Docking. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2018, 38(02): 654-659.
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