冀旭 孙芳云 赵勤 张晓英 李岩松

【摘 要】 目的：以栎瘿酸为例，研究藏药有效成分与功能蛋白的相互作用与识别过程。方法：利用荧光发射光谱、同步荧光、紫外光谱等方法进行研究。结果：结果显示栎瘿酸引起了牛血清白蛋白BSA荧光静态猝灭;获得不同温度下二者的结合常数、结合位点数及结合过程中热力学参数;得知二者结合是以氢键和范德华力为主要作用力的自发、放热过程;二者之间发生非辐射能量转移，结合部位位于BSA分子Site Ⅱ位点，平均结合距离为4.134 nm;在识别过程中，BSA结构及色氨酸所处微环境疏水性增加，色胺酸残基流动性增强。结论：通过本方法可建立一套用于表征藏药中有效成分与功能蛋白之间相互作用与识别过程的实验方法。

【关键词】 藏药;有效成分;功能蛋白;秦艽;栎瘿酸

【中图分类号】R917   【文献标志码】 A    【文章编号】1007-8517（2021）12-0054-07

Abstract：Objective Taking Roburic acid as an example，the interaction and recognition process between effective constituents of Tibetan medicine and functional proteins were studied in this paper. Methods Fluorescence emission spectroscopy，synchronous fluorescence，ultraviolet spectroscopy and other methods were used in this study. Result The result showed that，firstly，the binding procedures between Roburic acid and BSA were static quenching process caused by the formation of non-fluorescigenic complex; secondly，both binding constants and binding site numbers of Roburic acid and BSA at different temperatures were determined; thirdly，ΔG，ΔH and ΔS during the binding process can be obtained and all of these parameters were negative，which indicated the binding procedure between Roburic acid and BSA was spontaneous，exothermic reaction taking hydrogen bond and van der waals force as main force; fourthly，“Forster non-radiative energy transfer” was taken place between Roburic acid and BSA，and Roburic acid bound to the Site Ⅱ of BSA while the binding distances was 4.134nm; fifthly，during the recognition procedure，with the increasing concentration of Roburic acid，the hydrophobicity of both BSA structure and the microenvironment of tryptophan in BSA increased，and the tryptophan residues were no longer restricted.Conclusion This study can be used to establish a set of experimental methods to characterize the interaction and recognition process between traditional Tibetan medicine effective constituents and functional protein.

Keywords：Tibetan Medicine; Effective Constituent; Functional Protein; Gentianae macrophyllae Radix; Roburic Acid

在生物學中，分子间相互作用与识别是形成高度专一性识别、反应及调控等过程的基础，诸如底物与受体的结合识别、酶反应及抗体-抗原的结合识别等。而在药学，尤其是民族药的开发研究领域，分子间相互作用也是一个重要的研究内容，它在基于传统药物的新药筛选开发、药物药效学及药动学研究中均得到广泛应用。藏药在我国有着悠久的应用历史、深厚的民众基础及丰富的藏民族文化内涵。近年来，藏药以其独特的功效受到越来越多研究者的关注，对其的现代化研究与应用也日益得到重视。而关于藏药当中有效成分与离子通道、膜蛋白等药物受体、功能蛋白的相互作用与识别研究还未见报道。……