Selective superoxide generation within mitochondria by the targeted redox cycler MitoParaquat
Selective superoxide generation within mitochondria by the targeted redox cycler MitoParaquat
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DOI：
10.1016/j.freeradbiomed.2015.08.021
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发表时间：
2015-12-01
影响因子：
7.4
通讯作者：
Murphy, Michael P.
中科院分区：
文献类型：
--
作者：
Robb, Ellen L.;Gawel, Justyna M.;Murphy, Michael P.
超氧化物是线粒体呼吸链产生的近端活性氧 (ROS)，在病理性氧化应激和氧化还原信号传导中发挥重要作用。虽然有一些工具可以检测或减少线粒体超氧化物，但没有一种工具可以快速、特异性地增加线粒体基质内超氧化物的产生。这种缺乏阻碍了进展，使得准确评估线粒体超氧化物在细胞和体内的作用具有挑战性。为了解决这一未满足的需求，我们合成并表征了一种线粒体靶向氧化还原循环剂 MitoParaquat (MitoPQ)，它包含与氧化还原循环剂百草枯缀合的三苯基鏻亲脂性阳离子。 MitoPQ 在膜电位的驱动下选择性地积聚在线粒体基质中。在基质内，MitoPQ 通过复合物 I 黄素位点的氧化还原循环产生超氧化物，选择性地增加线粒体内超氧化物的产生。 MitoPQ 增加分离线粒体和培养细胞中线粒体超氧化物的效果比非靶向百草枯高千倍。在离体灌注心脏和果蝇体内，MitoPQ 的毒性也比百草枯更高。 MitoPQ 能够在线粒体内选择性生成超氧化物，是研究线粒体超氧化物在细胞和体内病理学和氧化还原信号传导中的多种作用的有用工具。 (C) 2015 年作者。
Superoxide is the proximal reactive oxygen species (ROS) produced by the mitochondrial respiratory chain and plays a major role in pathological oxidative stress and redox signaling. While there are tools to detect or decrease mitochondrial superoxide, none can rapidly and specifically increase superoxide production within the mitochondrial matrix. This lack impedes progress, making it challenging to assess accurately the roles of mitochondrial superoxide in cells and in vivo. To address this unmet need, we synthesized and characterized a mitochondria-targeted redox cycler, MitoParaquat (MitoPQ) that comprises a triphenylphosphonium lipophilic cation conjugated to the redox cycler paraquat. MitoPQ accumulates selectively in the mitochondrial matrix driven by the membrane potential. Within the matrix, MitoPQ produces superoxide by redox cycling at the flavin site of complex I, selectively increasing superoxide production within mitochondria. MitoPQ increased mitochondrial superoxide in isolated mitochondria and cells in culture a thousand-fold more effectively than untargeted paraquat. MitoPQ was also more toxic than paraquat in the isolated perfused heart and in Drosophila in vivo. MitoPQ enables the selective generation of superoxide within mitochondria and is a useful tool to investigate the many roles of mitochondrial superoxide in pathology and redox signaling in cells and in vivo. (C) 2015 The Authors.