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首页> 外文期刊>Progress in Artificial Intelligence >Visualization of the Redox Status of Cytosolic Glutathione Using the Organelle- and Cytoskeleton-Targeted Redox Sensors
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Visualization of the Redox Status of Cytosolic Glutathione Using the Organelle- and Cytoskeleton-Targeted Redox Sensors

机译：使用细胞器和细胞骨架靶向氧化还原传感器可视化细胞骨谷胱甘肽的氧化还原状态

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摘要

Glutathione is a small thiol-containing peptide that plays a central role in maintaining cellular redox homeostasis. Glutathione serves as a physiologic redox buffer by providing thiol electrons for catabolizing harmful oxidants and reversing oxidative effects on biomolecules. Recent evidence suggests that the balance of reduced and oxidized glutathione (GSH/GSSG) defines the redox states of Cys residues in proteins and fine-tunes their stabilities and functions. To elucidate the redox balance of cellular glutathione at subcellular resolution, a number of redox-sensitive green fluorescent protein (roGFP) variants have been developed. In this study, we constructed and functionally validated organelle- and cytoskeleton-targeted roGFP and elucidated the redox status of the cytosolic glutathione at a subcellular resolution. These new redox sensors firmly established a highly reduced redox equilibrium of cytosolic glutathione, wherein significant deviation was observed among cells. By targeting the sensor to the cytosolic and lumen sides of the Golgi membrane, we identified a prominent redox gradient across the biological membrane at the Golgi body. The results demonstrated that organelle- and cytoskeleton-targeted sensors enable the assessment of glutathione oxidation near the cytosolic surfaces of different organelle membranes.
机译：谷胱甘肽是一种含小硫醇的肽，其在维持细胞氧化还原稳态中起着重要作用。通过提供用于分解有害氧化剂并逆转生物分子的氧化作用来用作生理氧化还原缓冲液。最近的证据表明，减少和氧化的谷胱甘肽（GSH / GSSG）的平衡定义了蛋白质中Cys残留物的氧化还原状态，并微调其稳定性和功能。为了阐明在亚细胞分辨率下细胞谷胱甘肽的氧化还原平衡，已经开发了许多氧化还原敏感的绿色荧光蛋白（ROGFP）变体。在这项研究中，我们构建和功能验证的细胞内细胞石和细胞骨架靶向RogFP，并以亚细胞分辨率阐明了细胞溶质谷胱甘肽的氧化还原状态。这些新的氧化还原传感器牢固地建立了一种高度减少的细胞溶质谷胱甘肽的氧化还原平衡，其中细胞之间观察到显着偏差。通过将传感器靶向高尔基膜的细胞溶质和腔侧，我们在高尔基体上识别了整个生物膜的突出氧化还原梯度。结果表明，细胞器和细胞骨架靶向传感器使得能够评估不同细胞内膜的细胞骨表面附近的谷胱甘肽氧化。

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