物理化学学报 >> 2009, Vol. 25 >> Issue (10): 2147-2154.doi: 10.3866/PKU.WHXB20091112

研究论文 上一篇    下一篇

光谱法测定伊曲康唑与牛血清和人血清白蛋白相互作用

郭清莲, 李冉, 蒋风雷, 涂建成, 李林尉, 刘义   

  1. 武汉大学中南医院, 武汉 430071|武汉大学化学与分子科学学院, 病毒学国家重点实验室及分析生物医学教育部重点实验室, 武汉 430072|聊城大学化学化工学院, 山东 聊城 252059
  • 收稿日期:2009-07-20 修回日期:2009-08-28 发布日期:2009-09-29
  • 通讯作者: 涂建成 E-mail:Jian_1999@yahoo.com

Characterization of the Interactions between Itraconazole and Human and Bovine Serum Albumins by a Spectroscopic Method

GUO Qing-Lian, LI Ran, JIANG Feng-Lei, TU Jian-Cheng, LI Lin-Wei, LIU Yi   

  1. Zhongnan Hospital, Wuhan University, Wuhan 430071, P. R. China|State Key Laboratory of Virology &|Key Laboratory of Analytic Chemistry for Biology and Medicine of Ministry of Education, College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, P. R. China|College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng 252059, Shandong Province, P. R. China
  • Received:2009-07-20 Revised:2009-08-28 Published:2009-09-29
  • Contact: TU Jian-Cheng E-mail:Jian_1999@yahoo.com

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摘要：

用荧光光谱和紫外吸收光谱法, 在pH=7.4±0.1的0.1 mol·L-1磷酸缓冲溶液中, 研究了伊曲康唑与牛血清白蛋白(BSA)和人血清白蛋白(HSA)的相互作用. 实验结果表明, 伊曲康唑与牛血清白蛋白和人血清白蛋白作用的猝灭常数均随着温度的升高而降低, 伊曲康唑可以有规律地使血清白蛋白内源荧光猝灭, 其猝灭机理可认为是伊曲康唑与白蛋白形成复合物的静态猝灭. 获得了在不同温度下, 伊曲康唑与血清白蛋白作用的结合常数以及△G、△H和△S等热力学参数. 根据所得结果可推断伊曲康唑与白蛋白的作用力主要为疏水作用力, 同时, 利用荧光共振能量转移理论(FRET)计算得出了伊曲康唑与白蛋白结合位置的距离d. 而且, 利用同步荧光光谱和紫外光谱揭示了该反应中蛋白的结构和其微环境的变化.

关键词: 热力学参数, 伊曲康唑, 牛血清白蛋白, 人血清白蛋白, 荧光猝灭, 紫外可见光谱

Abstract:

The binding of itraconazole (ITZ), a potential antifungal, to human serumalbumin (HSA) and bovine serum albumin (BSA) were studied at the physiological acidity (pH=7.4±0.1) by fluorescence and UV-Vis spectroscopies. A decrease in the quenching constant was observed with an increase in temperature. From the fluorescence spectrum and the fluorescence intensity, we observed that ITZ strongly quenches the intrinsic fluorescence of both BSA and HSA by static quenching. Thermodynamic parameters, such as △G, △H and △S, were calculated at different temperatures, showing that electrostatic and hydrophobic interactions were mostly responsible for the binding of ITZ to serum albumin. The distance d between the donor (HAS or BSA ) and acceptor (ITZ) was obtained according to fluorescence resonance energy transfer theory (FRET). Synchronous fluorescence and UV-Vis spectroscopy clearly revealed that the microenvironment and the conformation of serumalbumins changed during the binding reaction.

Key words: Thermodynamic parameter, Itraconazole, Bovine serumalbumin, Human serumalbumin, Fluorescence quenching, UV-Vis spectroscopy