光谱法结合分子对接研究沙拉沙星与牛血清白蛋白结合作用

doi:10.16431/j.cnki.1671-7236.2025.09.035

摘要/Abstract

摘要： 【目的】研究沙拉沙星(salafloxacin，SFX)与牛血清白蛋白(bovine serum albumin，BSA)的结合作用机理。【方法】采用荧光光谱法测定SFX对BSA的荧光猝灭作用，探究SFX与BSA作用类型、结合方式、结合位点数；通过同步荧光法测定SFX对BSA内源性荧光作用，分析SFX对BSA中的酪氨酸(tyrosine，Tyr)和色氨酸(tryptophan，Trp)荧光强度的影响；通过三维荧光光谱法检测SFX和BSA的三维荧光特征，分析SFX对BSA构象的影响；采用紫外可见光谱法检测添加不同浓度SFX时BSA的紫外光谱特征，分析SFX对BSA构象的影响和猝灭类型；利用分子对接分析SFX与BSA结合位点和结合作用力；利用荧光光谱法分析金属离子对SFX和BSA结合的影响。【结果】荧光光谱检测结果显示，SFX能猝灭BSA的内源性荧光，对BSA猝灭类型为静态猝灭；SFX与BSA结合过程是放热、自发的，结合过程发生了非辐射能量转移；SFX和BSA结合的作用力是氢键和范德华力，结合位点数约为1，分子结合距离为3.182 nm。同步荧光检测结果显示，SFX猝灭了BSA中Tyr残基和Trp残基以及肽链骨架结构的荧光。三维荧光光谱检测结果显示，SFX引起Trp和Tyr残基的荧光强度降低，导致BSA肽链结构改变。分子对接分析结果显示，SFX与BSA中的Pro498、Glu470、Lys533残基形成氢键，与Val497、Lys499残基形成疏水键，与Tyr496残基形成范德华作用力。金属离子Ca²⁺、Na⁺、K⁺、Al^3＋对SFX与BSA结合有促进作用；Fe^2＋、Mg^2＋、Cu^2＋表现为抑制作用。【结论】 SFX和BSA结合是自发的,形成稳定的复合物,且结合会改变BSA的构象。试验结果为揭示SFX毒性、机体内转运和代谢研究提供数据支持。

关键词: 牛血清白蛋白(BSA); 沙拉沙星(SFX); 光谱法; 分子对接; 金属离子

Abstract: 【Objective】 The purpose of this experiment was to study the binding mechanism of salafloxacin (SFX) and bovine serum albumin (BSA). 【Method】 The fluorescence spectroscopy method was employed to investigate the fluorescence quenching effect of SFX on BSA,the interaction type,binding mode,and number of binding sites between SFX and BSA.Synchronous fluorescence spectroscopy was used to measure the effect of SFX on the endogenous fluorescence of BSA,the influence of SFX on the fluorescence intensity of tyrosine (Tyr) and tryptophan (Trp) in BSA was analyzed.The three-dimensional fluorescence spectroscopy was employed to analyze the three-dimensional fluorescence characteristics of SFX and BSA,and the effect of SFX on the conformation of BSA was analyzed.Ultraviolet-visible spectroscopy was employed to detect the UV spectral characteristics of BSA upon addition of varying concentrations of SFX，the effect of SFX on the conformation of BSA and the type of quenching was analyzed.The binding sites and binding forces between SFX and BSA were analyzed by molecular docking method，and the effect of metal ions on the binding of SFX and BSA was analyzed by fluorescence spectroscopy. 【Result】 Fluorescence spectroscopy detection results showed that SFX quenched the endogenous fluorescence of BSA,and the quenching type of BSA was static quenching.The binding process between SFX and BSA was exothermic and spontaneous，non radiative energy transfer occurred during the binding process.The interaction between SFX and BSA was primarily mediated by hydrogen bonding and van der Waals forces.The number of binding sites was approximately 1.The molecular binding distance was 3.182 nm.Synchronous fluorescence detection showed that SFX quenched the fluorescence of Tyr and Trp residues as well as the peptide backbone in BSA.Three-dimensional fluorescence spectroscopy demonstrated a decrease in fluorescence intensity of Trp and Tyr residues induced by SFX,and the peptide chain structure of BSA was alterated.Molecular docking showed that SFX formed hydrogen bonds with Pro498,Glu470 and Lys533 residues in BSA,hydrophobic bonds with Val497 and Lys499 residues,and van der Waals forces with Tyr496 residue.The metal ions Ca²⁺,Na⁺,K⁺ and Al³⁺exhibited a promoting effect on the binding of SFX to BSA，Fe²⁺,Mg²⁺ and Cu²⁺ exhibited inhibitory effects. 【Conclusion】 SFX-BSA binding was spontaneous,forming a stable complex,and the combination would change the conformation of BSA.This study provided fundamental data for investigating SFX toxicity,in vivo transport and metabolic mechanisms.

Key words: bovine serum albumin (BSA); salafloxacin (SFX); spectral method; molecular docking; metal ion

中图分类号:

S859.7

马全朝, 吴海港. 光谱法结合分子对接研究沙拉沙星与牛血清白蛋白结合作用[J]. 中国畜牧兽医, 2025, 52(9): 4394-4405.

MA Quanchao, WU Haigang. Study on the Interaction Between Salafloxacin and Bovine Serum Albumin by Spectroscopic Method Combined with Molecular Docking[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(9): 4394-4405.

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