色谱

色谱

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高效亲和色谱法测定丹皮酚与固定化人血清白蛋白结合域

于雪1, 张君才1,2, 卫引茂1*   

  1. 1. 西北大学合成与天然功能分子化学教育部重点实验室, 化学与材料科学学院, 陕西 西安 710069; 2. 咸阳师范学院化学与化工学院, 陕西 咸阳 750021
  • 收稿日期:2010-03-21 修回日期:2010-05-21 出版日期:2010-07-28 发布日期:1980-03-25
  • 通讯作者: 卫引茂

Determination of binding domains of paeonol on immobilized human serum albumin by high-performance affinity chromatography

YU Xue1, ZHANG Juncai1,2, WEI Yinmao1*   

  1. 1. Key Laboratory of Synthetic and Natural Function Molecule Chemistry of Ministry of Education, College of Chemistry & Material Science, Northwest University, Xi’an 710069, China; 2. School of Chemistry and Chemical Engineering, Xianyang Normal University, Xianyang 750021, China
  • Received:2010-03-21 Revised:2010-05-21 Online:2010-07-28 Published:1980-03-25
  • Contact: WEI Yinmao

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摘要： 利用亲和色谱,在模拟人体生理环境下(37 ℃、pH 7.4),采用竞争置换法研究了丹皮酚(PAE)与固定化人血清白蛋白(HSA)的相互作用。通过对PAE的自我竞争分析及PAE与HSA上结合位点的标记物间的竞争置换分析,得到了PAE和HSA间的结合常数、结合位点数和结合域。结果表明: PAE在HSA分子中仅存在一类结合位点,结合常数为4.84×103 L/mol,该结合位点为HSA上的Sudlow siteII;通过对PAE与HSA相互作用的热力学研究,推断出二者间的作用力类型为氢键或范德华力。

关键词: 丹皮酚, 高效亲和色谱, 竞争置换分析, 人血清白蛋白

Abstract: High-performance affinity chromatography was used to investigate the binding of paeonol (PAE) to immobilized human serum albumin (HSA) under the condition of pH 7.4 and temperature of 37 ℃. Self-competition zonal elution indicated that there was only one major binding site on HSA for PAE. The association constant of PAE with HSA was 4.84×103 L/mol. Competition studies based on zonal elution were carried out between PAE and various probes which have been known to interact with several major and minor sites on HSA. PAE was found to have direct competition with L-tryptophan. The results indicated that PAE interacted with Sudlow site II on HSA. The thermodynamic analysis indicated that the main force between the paeonol and HSA was hydrogen bond or van der Waals force.

Key words: competition and displacement studies, human serum album (HSA) , paeonol, high-performance affinity chromatography