Hydrogen peroxide probes directed to different cellular compartments.
Hydrogen peroxide probes directed to different cellular compartments.
复制标题
过氧化氢探针针对不同的细胞室。
DOI：
10.1371/journal.pone.0014564
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发表时间：
2011-01-21
期刊：
影响因子：
3.7
通讯作者：
Gladyshev VN
中科院分区：
文献类型：
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作者：
Malinouski M;Zhou Y;Belousov VV;Hatfield DL;Gladyshev VN
过氧化氢的可控生成和清除在细胞氧化还原、动态平衡和信号转导中起着重要作用。我们使用了针对不同隔室的过氧化氢生物传感器HYPER来检测哺乳动物细胞中的这些过程。观察到对各种氧化还原扰动和信号事件的可逆响应。在HEK 293细胞中的高表达被发现能感觉到低微摩尔水平的过氧化氢。当针对不同的细胞隔室时，在细胞核、胞浆、过氧化体、线粒体膜间隙和线粒体基质中出现高水平的还原状态，但在这些隔室中也观察到低水平的氧化形式的生物传感器，这与哺乳动物细胞的低过氧化值相一致。相反，HYPER主要在内质网上被氧化。使用这个系统，我们描述了不同细胞系统中过氧化氢的控制，例如缺乏硫氧还蛋白还原酶、硫氧还蛋白还原酶、硫氧还蛋白还原酶或处于缺硒状态的细胞。过氧化氢的产生也可以在信号事件发生后的不同隔间中进行监测。我们发现，HYPER可以作为一种有价值的工具来监测不同细胞隔间产生的过氧化氢。数据还表明，在一个隔间产生的过氧化氢可能会转移到其他隔间。我们的数据提供了关于哺乳动物细胞中过氧化氢的区隔、动态和动态平衡控制的信息。
Controlled generation and removal of hydrogen peroxide play important roles in cellular redox homeostasis and signaling. We used a hydrogen peroxide biosensor HyPer, targeted to different compartments, to examine these processes in mammalian cells. Reversible responses were observed to various redox perturbations and signaling events. HyPer expressed in HEK 293 cells was found to sense low micromolar levels of hydrogen peroxide. When targeted to various cellular compartments, HyPer occurred in the reduced state in the nucleus, cytosol, peroxisomes, mitochondrial intermembrane space and mitochondrial matrix, but low levels of the oxidized form of the biosensor were also observed in each of these compartments, consistent with a low peroxide tone in mammalian cells. In contrast, HyPer was mostly oxidized in the endoplasmic reticulum. Using this system, we characterized control of hydrogen peroxide in various cell systems, such as cells deficient in thioredoxin reductase, sulfhydryl oxidases or subjected to selenium deficiency. Generation of hydrogen peroxide could also be monitored in various compartments following signaling events. We found that HyPer can be used as a valuable tool to monitor hydrogen peroxide generated in different cellular compartments. The data also show that hydrogen peroxide generated in one compartment could translocate to other compartments. Our data provide information on compartmentalization, dynamics and homeostatic control of hydrogen peroxide in mammalian cells.