SLC25A39 is necessary for mitochondrial glutathione import in mammalian cells
SLC25A39 is necessary for mitochondrial glutathione import in mammalian cells
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DOI：
10.1038/s41586-021-04025-w
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发表时间：
2021-10-27
期刊：
影响因子：
64.8
通讯作者：
Birsoy, Kivanc
中科院分区：
文献类型：
--
作者：
Wang, Ying;Yen, Frederick S.;Birsoy, Kivanc
谷胱甘肽（GSH）是一种小分子硫醇，在所有真核生物中含量丰富，在氧化代谢中起关键作用（1）。线粒体作为氧化反应的主要场所，必须保持足够的GSH水平以执行保护和生物合成功能（2）。GSH只在胞质溶胶中合成，但线粒体GSH输入所涉及的分子机制仍然未知。在这里，使用细胞器蛋白质组学和代谢组学的方法，我们确定SLC 25 A39，一个线粒体膜载体的未知功能，作为一个调节剂的GSH运输到线粒体。SLC 25 A39的缺失减少了线粒体GSH的输入和丰度，而不影响细胞GSH水平。同时缺乏SLC 25 A39和它的副产物SLC 25 A40的细胞表现出含有铁-硫簇的蛋白质的活性和稳定性的缺陷。我们发现，线粒体GSH输入是必要的细胞增殖在体外和小鼠红细胞发育。工程化的双功能细菌GSH生物合成酶（GshF）在线粒体中的异源表达使线粒体GSH产生成为可能并改善由其耗尽引起的代谢和增殖缺陷。最后，GSH的可用性负调控SLC 25 A39蛋白丰度，耦合氧化还原稳态线粒体GSH输入哺乳动物细胞。我们的工作确定SLC 25 A39作为一个重要的和受管制的组成部分，线粒体GSH进口机器。
Glutathione (GSH) is a small-molecule thiol that is abundant in all eukaryotes and has key roles in oxidative metabolism(1). Mitochondria, as the major site of oxidative reactions, must maintain sufficient levels of GSH to perform protective and biosynthetic functions(2). GSH is synthesized exclusively in the cytosol, yet the molecular machinery involved in mitochondrial GSH import remains unknown. Here, using organellar proteomics and metabolomics approaches, we identify SLC25A39, a mitochondrial membrane carrier of unknown function, as a regulator of GSH transport into mitochondria. Loss of SLC25A39 reduces mitochondrial GSH import and abundance without affecting cellular GSH levels. Cells lacking both SLC25A39 and its paralogue SLC25A40 exhibit defects in the activity and stability of proteins containing iron-sulfur clusters. We find that mitochondrial GSH import is necessary for cell proliferation in vitro and red blood cell development in mice. Heterologous expression of an engineered bifunctional bacterial GSH biosynthetic enzyme (GshF) in mitochondria enables mitochondrial GSH production and ameliorates the metabolic and proliferative defects caused by its depletion. Finally, GSH availability negatively regulates SLC25A39 protein abundance, coupling redox homeostasis to mitochondrial GSH import in mammalian cells. Our work identifies SLC25A39 as an essential and regulated component of the mitochondrial GSH-import machinery.