CAJ | 학술논문

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T RX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T RX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T RX氧化态‐还原态之间的相互转化。结果表明氧化态的T RX与还原态的G PX3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G PX3将电子传递给T RX ，为揭示T RX在细胞信号传递过程中的物理化学机制提供了实验依据。
세포내단백질적양화환원상태직접영향세포적증식、분화급조망，이양화환원상태적개변대조공세포적생존혹사망우위중요。류양환단백（Thioredoxin ，TRX）시일충엄범존재우생물체내적양화환원조절단백，기재세포내양화환원상태적변화시발휘기양화환원조공작용적중요과정。이T RX위대상병이기중적색안산잔기（Trp）작위내품형광탐침，이용단백질정점돌변、SDS‐취병희선알응효전영（SDS‐PAGE）、형광광보화원이색보등기술화방법，연구TRX여곡광감태과양화물매（glutathione peroxidase ，GPX3）상호작용과정중양화환원태적변화。통과관측T RX이급돌변체중색안산형광광보적변화，연구단백상호작용적전자전이모식이급T RX양화태‐환원태지간적상호전화。결과표명양화태적T RX여환원태적G PX3지간존재상호작용병발생전자교환，해석료이자지간전자전체모식위G PX3장전자전체급T RX ，위게시T RX재세포신호전체과정중적물이화학궤제제공료실험의거。
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 .