广西师范大学学报(自然科学版) ›› 2014, Vol. 32 ›› Issue (2): 82-87.

• • 上一篇    下一篇

溴酚蓝与牛血清白蛋白的相互作用研究

邹华, 周享春, 孙梅香, 王玉龙   

  1. 长江大学化学与环境工程学院,湖北荆州434023
  • 收稿日期:2014-02-24 出版日期:2014-06-25 发布日期:2018-09-25
  • 通讯作者: 周享春(1962—),男,湖北荆州人,长江大学教授。E-mail:xiangchunzhou@163.com
  • 基金资助:
    国家自然科学基金资助项目(21201023)

Interaction of Bromophenol Blue and Bovine Serum Albumin

ZOU Hua, ZHOU Xiang-chun, SUN Mei-xiang, WANG Yu-long   

  1. College of Chemistry and Environmental Engineering,Yangtze University,Jingzhou Hubei 434023,China
  • Received:2014-02-24 Online:2014-06-25 Published:2018-09-25

摘要: 在模拟动物体生理条件和不同温度下,采用荧光、紫外光谱法研究溴酚蓝(BPB)与牛血清白蛋白(BSA)结合反应。用Stern-Volmer和Lineweaver-Burk方程分别处理试验数据,发现BSA 与BPB发生反应,生成新的复合物,属于静态荧光猝灭。实验求出反应时复合物的形成常数KLB、热力学参数与结合位点数,证明溴酚蓝与牛血清白蛋白的结合主要为静电作用力。位点竞争实验结果显示BPB与BSA的作用位置主要在BSA的Site Ⅰ(sub-domain ⅡA)位。通过同步荧光光谱和三维荧光光谱法,探讨BPB对BSA 构象的影响,实验结果表明BPB使色氨酸残基所处微环境的极性减弱、疏水作用增强。本文为探讨BPB的染色机理、毒理效应和生物学效应提供了重要信息。

关键词: 溴酚蓝, 牛血清白蛋白, 荧光猝灭光谱, 三维荧光光谱, 热力学参数

Abstract: Under simulated physiological conditions of animals and at various temperatures, the binding reaction of Bromophenol blue (BPB) to bovine serum albumin (BSA) was studied by fluorescence spectrum and ultra-violet spectrum spectroscopy. The fluorescence quenching data were analyzed according to Stem-Volmer equation and Lineweaver-Burk double-reciprocal equation. The study show that BSA reacted with BPB and formed a certain new compound, which belonged to static fluorescence quenching. The formation constants of the compound KLB, the thermodynamic parameters and the number of binding sites were obtained. And the binding power between them is mainly the electrostatic acting force. Site marker competitive experiments showed that the binding of BPB to BSA primarily took place in site Ⅰ(sub-domain ⅡA)of BSA. The effect of BPB on the conformation of BSA was analyzed by synchronous fluorescence spectra and three-dimensional fluorescence spectra, which indicates that the polarity microenvironment around Trp residues decreased, while hydrophobic forces increased. These provide important information for enucleating the dyeing mechanisms, the toxicity effects and biological effects of BPB.

Key words: bromophenol blue, bovine serum albumin, fluorescence quenching spectrum, three dimensional fluorescence spectrum, thermodynamic parameters

中图分类号: 

  • Q512+.1
[1] 颜承农,张华新,刘义,等.百草枯与牛血清白蛋白结合作用的荧光光谱[J].化学学报,2005,63(18):1727-1732.
[2] HU Yan-jun, LIU Yi, XIAO Xiao-he. Investigation of the interaction between berberine and human serum albumin[J]. Biomacromolecules, 2009, 10: 517-521.
[3] 周享春,陈有胜.二乙酰乙肟与牛血清白蛋白相互作用的研究[J]. 计算机与应用化学,2011,28(3):320-324.
[4] 周享春,金小芬,宁方香,等.山梨酸钾与牛血清白蛋白相互作用的研究[J].中国食品添加剂,2010(103):107-113.
[5] 周享春,颜承农,王特,等.罗丹明B与牛血清白蛋白的相互作用研究[J].化学研究与应用,2009,21(1):36-41.
[6] ROSS D P, SUBRAMANIAN S. Thermodynamics of protein association reaction:Forces contributing to stability[J]. Biochemistry, 1981, 20: 3096-3099.
[7] 颜承农,童金强,熊丹. 荧光光谱法研究培氟沙星与牛血清白蛋白结合反应特征[J]. 分析化学,2006,34(6):796-800.
[8] 周享春, 周玉琴. 对硝基氯苯与牛血清白蛋白的相互作用研究[J]. 化学试剂, 2010, 32(9): 774-778.
[9] 梁宏. 血清白蛋白的构象及构象变化的研究进展[J]. 广西师范大学学报: 自然科学版, 1994, 12(1):54-59.
[10] 周享春,颜昌琪,颜承农.溴百里酚蓝与牛血清白蛋白的相互作用研究[J].化学研究与应用,2010,22(1):18-23.
No related articles found!
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 广西师范大学学报(自然科学版)编辑部
地址:广西桂林市三里店育才路15号 邮编:541004
电话:0773-5857325 E-mail: gxsdzkb@mailbox.gxnu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发