Development of novel FP-based probes for live-cell imaging of nitric oxide dynamics.
Development of novel FP-based probes for live-cell imaging of nitric oxide dynamics.
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DOI：
10.1038/ncomms10623
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发表时间：
2016-02-04
影响因子：
16.6
通讯作者：
Malli R
中科院分区：
文献类型：
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作者：
Eroglu E;Gottschalk B;Charoensin S;Blass S;Bischof H;Rost R;Madreiter-Sokolowski CT;Pelzmann B;Bernhart E;Sattler W;Hallström S;Malinski T;Waldeck-Weiermair M;Graier WF;Malli R
一氧化氮（）是一种具有广泛生物学效应的自由基，但在单个细胞中几乎不可能可视化。在这里，我们报告了新的多色荧光猝灭为基础的探针融合细菌衍生的结合结构域接近不同的荧光蛋白变体的发展。这些基因编码的探针，被称为geNOps，提供了一个选择性，特异性和实时读出的细胞动力学，因此，打开了一个新的时代的生物成像。geNOps与Ca2+传感器的组合使我们能够在单个内皮细胞中同时观察和Ca2+信号。此外，探针的靶向用于检测线粒体内的信号。geNOps是进一步研究和理解单（亚）细胞水平上信号传导的复杂模式的有用新工具。 一氧化氮是一种挥发性自由基第二信使，具有多种生物学效应。在这里，Eroglu等人开发了一氧化氮的遗传编码荧光生物传感器，并使用它们来可视化单细胞中的亚细胞一氧化氮动力学。
Nitric oxide () is a free radical with a wide range of biological effects, but practically impossible to visualize in single cells. Here we report the development of novel multicoloured fluorescent quenching-based probes by fusing a bacteria-derived -binding domain close to distinct fluorescent protein variants. These genetically encoded probes, referred to as geNOps, provide a selective, specific and real-time read-out of cellular dynamics and, hence, open a new era of bioimaging. The combination of geNOps with a Ca2+ sensor allowed us to visualize and Ca2+ signals simultaneously in single endothelial cells. Moreover, targeting of the probes was used to detect signals within mitochondria. The geNOps are useful new tools to further investigate and understand the complex patterns of signalling on the single (sub)cellular level. Nitric oxide is a volatile free radical second messenger with a large number of biological effects. Here Eroglu et al. develop genetically encoded fluorescent biosensors for nitric oxide and use them to visualise subcellular nitric oxide dynamics in single cells.