Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide.
Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide.
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DOI：
10.1038/ncomms6222
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发表时间：
2014-10-21
影响因子：
16.6
通讯作者：
Belousov VV
中科院分区：
文献类型：
--
作者：
Ermakova YG;Bilan DS;Matlashov ME;Mishina NM;Markvicheva KN;Subach OM;Subach FV;Bogeski I;Hoth M;Enikolopov G;Belousov VV
活性氧（ROS）是许多细胞功能的保守调节因子，并且ROS的过度产生是各种病理过程的标志。基因编码的荧光探针是研究不同规模和复杂性的生命系统中ROS产生的独特工具。然而，目前可用的重组氧化还原传感器具有绿色发射，其与许多其他探针的光谱重叠。扩大重组体内ROS探针的光谱范围将使多参数体内ROS检测成为可能。在这里，我们提出了第一个基因编码的红色荧光传感器过氧化氢检测，HyperRed。该传感器的性能与其绿色类似物相似。我们证明了效用的传感器通过跟踪低浓度的H2O2在细胞质中产生的培养细胞生长因子刺激后。此外，使用HyperRed，我们检测到本地和瞬时过氧化氢的生产在线粒体基质抑制内质网Ca 2+摄取后。
Reactive oxygen species (ROS) are conserved regulators of numerous cellular functions, and overproduction of ROS is a hallmark of various pathological processes. Genetically encoded fluorescent probes are unique tools to study ROS production in living systems of different scale and complexity. However, the currently available recombinant redox sensors have green emission, which overlaps with the spectra of many other probes. Expanding the spectral range of recombinant in vivo ROS probes would enable multiparametric in vivo ROS detection. Here we present the first genetically encoded red fluorescent sensor for hydrogen peroxide detection, HyPerRed. The performance of this sensor is similar to its green analogues. We demonstrate the utility of the sensor by tracing low concentrations of H2O2 produced in the cytoplasm of cultured cells upon growth factor stimulation. Moreover, using HyPerRed we detect local and transient H2O2 production in the mitochondrial matrix upon inhibition of the endoplasmic reticulum Ca2+ uptake.