Intracellular glutathione pools are heterogeneously concentrated.
Intracellular glutathione pools are heterogeneously concentrated.
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DOI：
10.1016/j.redox.2013.10.005
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发表时间：
2013
期刊：
影响因子：
11.4
通讯作者：
Appenzeller-Herzog, Christian
中科院分区：
文献类型：
--
作者：
Montero, Davide;Tachibana, Christine;Winther, Jakob Rahr;Appenzeller-Herzog, Christian
关键词：
谷氨酸在细胞中以毫摩尔浓度存在，但其在细胞隔室中的相对分布仍然难以捉摸。我们选择了内质网（ER）作为一个例子细胞器来研究特定的隔室谷胱甘肽水平。使用谷氧还蛋白传感器（sCGrx 1 pER），它快速，特异性地与还原型谷胱甘肽（GSH）-谷胱甘肽二硫化物（GSSG）的氧化还原对平衡与已知的平衡常数，我们表明，[GSH]：[GSSG]在完整的HeLa细胞的ER的比例小于7：1。考虑到先前确定的ER中[GSH]2：[GSSG]的值为83 mM，这转化为ER中的总谷胱甘肽浓度（[GStot]=[GSH]+2[GSSG]）大于15 mM。由于测量到整合的细胞内[GStot]约为7 mM，因此我们得出结论，在ER膜上存在[GStot]梯度。一个可能的稳态机制，胞质来源的谷胱甘肽被困在ER进行了讨论。我们提出了一个高[GStot]作为ER环境的区别特征相比，细胞外空间。1-Cys谷氧还蛋白的谷胱甘肽化状态是[GSH]：[GSSG]的读数。隔室特异性[GStot]由[GSH]：[GSSG]和[GSH]2：[GSSG]给出。[GStot]在ER中比在人细胞的胞质溶胶中更高。
Glutathione is present in millimolar concentrations in the cell, but its relative distribution among cellular compartments remains elusive. We have chosen the endoplasmic reticulum (ER) as an example organelle to study compartment-specific glutathione levels. Using a glutaredoxin sensor (sCGrx1pER), which rapidly and specifically equilibrates with the reduced glutathione (GSH)–glutathione disulfide (GSSG) redox couple with known equilibrium constant, we showed that the [GSH]:[GSSG] ratio in the ER of intact HeLa cells is less than 7:1. Taking into consideration the previously determined value for [GSH]2:[GSSG] in the ER of 83 mM, this translates into a total glutathione concentration in the ER ([GStot]=[GSH]+2[GSSG]) of greater than 15 mM. Since the integrated, intracellular [GStot] was measured as ~7 mM, we conclude the existence of a [GStot] gradient across the ER membrane. A possible homeostatic mechanism by which cytosol-derived glutathione is trapped in the ER is discussed. We propose a high [GStot] as a distinguishing feature of the ER environment compared to the extracellular space. Glutathionylation status of a 1-Cys glutaredoxin is a readout for [GSH]:[GSSG]. Compartment-specific [GStot] is given by [GSH]:[GSSG] and [GSH]2:[GSSG]. [GStot] is higher in the ER than in the cytosol of human cells.
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