利用内禀荧光探针研究蛋白相互作用中硫氧还蛋白的氧化还原态  

Monitoring the Redox States of Thioredoxin in Protein-Protein Interaction Using Intrinsic Fluorescence Probe

作　　者：王攀[1] 郭爱宇 常贯晓 冉霞[1] 张宇[2] 郭立俊[1] 

机构地区：[1]河南大学光物理研究所,物理与电子学院,河南开封475004 [2]河南大学植物逆境教育部重点实验室,河南开封475004

出　　处：《光谱学与光谱分析》2015年第10期2797-2801,共5页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(21373077);河南大学青年科研人才种子基金项目资助

摘　　要：细胞内蛋白质的氧化还原状态直接影响细胞的增殖、分化及凋亡,而氧化还原状态的改变对调控细胞的生存或死亡尤为重要。硫氧还蛋白(Thioredoxin,TRX)是一种广泛存在于生物体内的氧化还原调节蛋白,其在细胞内氧化还原状态的变化是发挥其氧化还原调控作用的重要过程。以TRX为对象并以其中的色氨酸残基(Trp)作为内禀荧光探针,利用蛋白质定点突变、SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)、荧光光谱和圆二色谱等技术和方法,研究TRX与谷胱甘肽过氧化物酶(glutathione peroxidase,GPX3)相互作用过程中氧化还原态的变化。通过观测TRX以及突变体中色氨酸荧光光谱的变化,研究蛋白相互作用的电子转移模式以及TRX氧化态-还原态之间的相互转化。结果表明氧化态的TRX与还原态的GPX3之间存在相互作用并发生电子交换,解释了二者之间电子传递模式为GPX3将电子传递给TRX,为揭示TRX在细胞信号传递过程中的物理化学机制提供了实验依据。The cellular redox states directly affect cell proliferation ,differentiation and apoptosis ,and the redox states changes is particularly important to the regulation of cell survival or death .Thioredoxin is a kind of oxidation regulatory protein which is widely exists in organisms ,and the change of redox states is also an important process in redox regulation .In this work ,we have used the site‐directed mutagenesis of protein ,SDS‐polyacrylamide gel electrophoresis fluorescence spectroscopy and circular dichroism etc .,to investigate redox states changes between TRX （E .coli） and glutathione peroxidase（GPX3） during their inter‐action .By observing the fluorescence spectra of TRX and its mutants ,we have studied the protein interactions as well as the red‐ox states switching between oxidation state TRX and the reduced state GPX3 .The results demonstrate the presence of interac‐tions and electron exchanges occurring between reduced GPX3 and oxidized TRX ,which is of significance for revealing the physi‐cal and chemical mechanism of TRX in intracellular signal transduction .

关 键 词：硫氧还蛋白 氧化还原态 蛋白相互作用 色氨酸残基 荧光光谱 

分 类 号：O433.4[机械工程—光学工程]