溴百里酚蓝与牛血清白蛋白的相互作用研究.pdf

第2卷第1期
化学研究与应用
Val. 22 No. 1
2010年1月
Chemical Research and Application
Jan.,2010
文章编号:1004-1656(2010)01-0018-06
溴百里酚蓝与牛血清白蛋白的相互作用研究
周享春”,颜昌琪,颜承农
(长江大学化学与环境工程学院,湖北荆州434023)
摘要:在模拟动物体生理条件和不同温度下,用荧光光谱和紫外-可见吸收光谱法研究了溴百里酚蓝(BTB)与
牛血清白蛋白(BSA)结合反应的光谱行为。用 Stem- Volmer和 Lineweaver-Buk方程分别处理试验数据,发现
BSA与BTB发生反应生成了新的复合物,属于静态荧光猝灭。求出了反应时复合物的形成常数Kx(2.792x
e mo
热力学参数(△HF=-20.24kJ?mor',AS°=37.22?K1,△G=-31.25J?mol')与结合位
点数(1.1578)。根据 Forster偶极-偶极非辐射能量转移理论计算出结合位置距离212位色氨酸残基2.60nm,
证明二者主要靠静电作用力结合。同时用同步荧光光谱和三维荧光光谱法探讨了BB对BSA构象的影响,
表明BTB使色氨酸戏基所处徵环境的极性减弱、疏水作用增强,为阿明BTB的染色机理、毒理效应和生物学
效应提供重要信息。
关績词:溴百里酚蓝;牛血清白蛋白;荧光猝灭光谱;三维荧光光谐;热力学参数
中图分类号:0629.73文献标识码:A
Study on the interaction of bromothymol blue and bovine serum albumin
ZIOU Xiang-chun", YAN Chang-qi, YAN Cheng-nong
College of Chemistry and Environmental Engineering, Yangtze University, Jingzhou 434023, China)
Abstract Under simulated physiological conditions of animals and at various temperatures, the binding reaction of bromothymol blue
(BTB to bovine serum albumin(BS) was studied by fluorescence spectrumand ultra-violet spectrum spectroscopy. The
fluorescence quenching data were analyzed according to Stern-volmer equation and Lineweaver-burk double-reciprocal equation
Studies have shown: BSA had reacted with BTB and formed a certain new compound, the quenching belonged to static fluorescence
quenching. The format on o as f e n (7 1 m e hemd ami aames(AH =20. 4J
mol',AS°=37.22J?K1,△G°=31.25.09k?mo'2) and the number of binding sites(1.1578) were obtained. Based on the
Forster s theory of non-radiation energy transfer, the istance between binding locality and tryptophan residue-212 was2.60 nm. It
shows that the binding power between them is mainly the elec ostatic acting force. The effect of BTB on the conformation of BSA
was analyzed by synchronous fluorescence spectra and three-dimensional Fluorescence spectra. It indicates that the polarity
microenvironment aound Tp residues decreased, hydrophobic forces increased. These provide important infomation for enucleating
the dyeing mechanisms, the toxicity effects and biological effects of BTB
Key words: bromothymol blue; bovine serum albumin; fluorescence quenching spectrum; three dimensional fluorescence spectrum
thermodynamic parameters
溴百里酚蓝( Bromothymol blue,简称BTB),是BSA相互作用为模型,探讨其光谱行为、结合反应
一种常用酸破指示剂和吸附指示剂,同时在光度机理和热力学特征,并用同步荧光光谱和三维荧
法分析中常用作显色剂们。但是BTB是一种常见光光谱技术考察了BTB对BSA构象的变化。对
的环境污染物。本实验以BTB作为荧光探针与明确BTB对蛋白质的染色机理及在动物体内的代
收稿日期:2009-01~13;修回日期:200904-19
基金项目:湖北省自然科学基金项日(2005ABA067,2004ABA104)资助
联系人简介:周享春(1962-),男,教授,主要从事分析化学的教学与研究。Email:xiangchunzhou'@163.com