H2O2 in plant peroxisomes: an in vivo analysis uncovers a Ca(2+)-dependent scavenging system.
H2O2 in plant peroxisomes: an in vivo analysis uncovers a Ca(2+)-dependent scavenging system.
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DOI：
10.1111/j.1365-313x.2010.04190.x
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发表时间：
2010-06-01
期刊：
影响因子：
--
通讯作者：
Lo Schiavo F
中科院分区：
文献类型：
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作者：
Costa A;Drago I;Behera S;Zottini M;Pizzo P;Schroeder JI;Pozzan T;Lo Schiavo F
氧化应激是生活在氧气世界中的所有细胞面临的主要挑战。在活性氧中，H2O2 是一种众所周知的有毒分子，如今被认为是多种信号通路的特定组成部分。为了深入了解 H2O2 在植物细胞中的作用，需要有可靠、特异和非侵入性的体内检测方法。因此，基因编码的H2O2传感器HyPer在植物细胞的不同亚细胞区室（例如细胞质和过氧化物酶体）中表达。此外，通过使用新的基于 GFP 的 Cameleon Ca2+ 指示剂 D3cpv-KVK-SKL（针对过氧化物酶体），我们证明在诱导细胞质 Ca2+ 增加后，过氧化物酶体腔内的 Ca2+ 也会增加。在烟草和抽苔前和抽苔后拟南芥植物的叶过氧化物酶体中进行了 HyPer 荧光比的分析。这些分析使我们能够证明体内过氧化物酶体内 Ca2+ 升高会刺激过氧化氢酶活性，从而提高过氧化物酶体 H2O2 清除效率。
Oxidative stress is a major challenge for all cells living in an oxygen-based world. Among reactive oxygen species, H2O2, is a well known toxic molecule and, nowadays, considered a specific component of several signalling pathways. In order to gain insight into the roles played by H2O2 in plant cells, it is necessary to have a reliable, specific and non-invasive methodology for its in vivo detection. Hence, the genetically-encoded H2O2 sensor HyPer was expressed in plant cells in different subcellular compartments such as cytoplasm and peroxisomes. Moreover, with the use of the new GFP-based Cameleon Ca2+ indicator, D3cpv-KVK-SKL, targeted to peroxisomes, we demonstrated that the induction of cytoplasmic Ca2+ increase is followed by Ca2+ rise in the peroxisomal lumen. The analyses of HyPer fluorescence ratios were performed in leaf peroxisomes of tobacco and pre- and post-bolting Arabidopsis plants. These analyses allowed us to demonstrate that an intraperoxisomal Ca2+ rise in vivo stimulates catalase activity, increasing peroxisomal H2O2 scavenging efficiency.