Interaction between picric acid and bovine serum albumin by fluorescence spectrometry
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摘要: 运用荧光和紫外光谱法并结合分子模拟技术研究了人体生理条件下苦味酸(Picric acid,简称PA)与牛血清白蛋白(BSA)的结合作用。实验结果表明,苦味酸对BSA的荧光猝灭作用较强,该猝灭属于生成复合物的静态猝灭。根据实验结果求得了不同温度下苦味酸与牛血清白蛋白结合作用的结合位点数和结合常数。根据van’t Hoff方程计算出的热力学参数,确定它们之间的相互作用力由以氢键和范德华力为主转变为以疏水作用力为主。根据Foster非辐射能量转移理论,求得苦味酸在距色氨酸残基距离3.14nm的位置与BSA结合。三种探针竞争标记药物实验表明,苦味酸主要结合在BSA上site I位点,分子模拟证实了该结合位点。同步荧光光谱结果表明苦味酸的加入使BSA的构象发生了变化。Abstract: The Interaction between picric acid ( PA) and bovine serum albumin ( BSA) was studied at the physiological p H by fluorescence and ( UV- vis) absorption spectroscopy, coupled with molecular modeling approach.The experimental results showed that there was a strong fluorescence quenching of BSA by PA. The probable quenching mechanism of fluorescence of BSA by PA was a static quenching by forming the BSA- PA complex. The binding constant and the number of binding sites of PA with BSA at different temperatures were obtained by fluorescence quenching method. According to the van 't Hoff equation, the thermodynamics parameters were calculated, which suggested that the binding pattern was considered from reflection of hydrogen bond and vander Waals forces to thydrophobic interaction. The binding locality was an area 3.14 nm away from tryptophan residue in BSA based on the Foster theory of non- radiation energy transfer. Three competitive sitemarkers revealed that PA was mainly located in the region of site I, molecular docking studies cinfirmed the binding site.Moreover, the results of synchronous fluorescence spectra demonstrated that the conformation of BSA was changed in the presence of PA.
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