基于荧光染料和牛血清白蛋白嵌合物对氧化还原状态的探测  

Detection of the redox status with an embedding complex of fluorescent dye and bovine serum albumin

作　　者：王鹏 周黄梅 李磊[2] 张三军[1,3] WANG Peng;ZHOU Huang-mei;LI Lei;ZHANG San-jun(State Key Laboratory of Precision Spectroscopy,East China Normal University,Shanghai 200062;School of Science,Jiangnan University,Wuxi 214122;Collaborative Innovation Centre of Extreme Optics,Shanxi University,Taiyuan 030006)

机构地区：[1]华东师范大学精密光谱科学与技术国家重点实验室,上海200062 [2]江南大学理学院,无锡214122 [3]山西大学极端光学协同创新中心,太原030006

出　　处：《分析试验室》2019年第11期1257-1262,共6页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金项目(11774096)资助

摘　　要：为制备一种与生物分子兼容性好并且具有较大动态检测范围的新型氧化还原探针,将氨苯乙烯类荧光分子4-[4-(二甲氨基)苯乙烯基]-1-甲基吡啶碘(p-DASPMI)嵌合到BSA中形成p-DASPMI-BSA嵌合体探针,采用稳态荧光光谱和时间分辨荧光光谱技术研究其与还原剂和氧化剂反应的荧光性质及相互作用的机理。实验表明,BSA能够感受环境的氧化还原状态,并通过p-DASPMI-BSA嵌合体探针的荧光变化来反映。在不同的氧化还原状态下,还原剂能使得探针的荧光强度增强3倍左右,平均寿命从1.47 ns增加到2.12 ns。而氧化剂使整个样品的荧光淬灭,平均寿命从2.15 ns减小到1.47 ns。时间分辨荧光数据表明氧化剂和还原剂会引起BSA的结构变化,进而影响进入BSA的p-DASPMI分子的数量,并最终影响整个体系的荧光性质。这个探针对氧化还原的动态检测范围能达到45倍左右,在研究细胞内大动态范围的氧化还原成像方面具有较好的应用前景。To prepare a new redox probe with good biocompatibility and large dynamic detection range,a chimera probe,p-DASPMI-BSA,was formed by embedding the aminostyrene fluorescence molecule p-DASPMI into BSA.The fluorescence properties and interaction mechanism of the probe with reducing or oxidizing agent were investigated by steady-state fluorescence spectroscopy and time-resolved fluorescence spectroscopy.Experiments confirmed that the fluorescence emission changes of pDASPMI-BSA probe can reflect the environmental redox states.Results showed that the reducing agent can enhance the fluorescence intensity about three times under different redox conditions,and the average lifetime increased from 1.47 ns to 2.12 ns.While the oxidizing agent quenched the fluorescence of the sample,the average lifetime also reduced from 2.15 ns to 1.47 ns.The fitting data indicated that the oxidizing or reducing agent cause the structural changes of BSA,which in turn affected the numbers of p-DASPMI molecules binding with BSA,and affected the fluorescence properties of the whole system.The dynamic detection range of this probe can reach to about 45 times,which has potential applications for imaging redox status of large scale in cells.

关 键 词：荧光探针 氧化还原状态 牛血清白蛋白 荧光光谱 

分 类 号：Q517[生物学—生物化学]