A DNA-based fluorescent probe maps NOS3 activity with subcellular spatial resolution.,Nature Chemical Biology

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A DNA-based fluorescent probe maps NOS3 activity with subcellular spatial resolution.
Nature Chemical Biology ( IF 12.9 ) Pub Date : 2020-03-09 , DOI: 10.1038/s41589-020-0491-3
Maulik S Jani _{1,

2} , Junyi Zou _{1,

2} , Aneesh T Veetil _{1,

2} , Yamuna Krishnan _{1,

2}

Affiliation

Nitric oxide synthase 3 (NOS3) produces the gasotransmitter nitric oxide (NO), which drives critical cellular signaling pathways by S-nitrosylating target proteins. Endogenous NOS3 resides at two distinct subcellular locations: the plasma membrane and the trans-Golgi network (TGN). However, NO generation arising from the activities of both these pools of NOS3 and its relative contribution to physiology or disease is not yet resolvable. We describe a fluorescent DNA-based probe technology, NOckout, that can be targeted either to the plasma membrane or the TGN, where it can quantitatively map the activities of endogenous NOS3 at these locations in live cells. We found that, although NOS3 at the Golgi is tenfold less active than at the plasma membrane, its activity is essential for the structural integrity of the Golgi. The newfound ability to spatially map NOS3 activity provides a platform to discover selective regulators of the distinct pools of NOS3.

中文翻译：

基于DNA的荧光探针可将NOS3活性与亚细胞空间分辨率对应起来。

一氧化氮合酶3（NOS3）产生气体递质一氧化氮（NO），该气体通过S-亚硝基化靶蛋白驱动关键的细胞信号通路。内源性NOS3位于两个不同的亚细胞位置：质膜和反高尔基体网络（TGN）。但是，由这两种NOS3库的活动及其对生理或疾病的相对贡献而产生的NO尚无法解决。我们描述了一种基于荧光DNA的探针技术，即NOckout，它既可以靶向质膜也可以靶向TGN，在那里它可以定量定位活细胞中这些位置的内源性NOS3的活性。我们发现，尽管高尔基体中的NOS3活性比质膜低十倍，但其活性对于高尔基体的结构完整性至关重要。

更新日期：2020-04-24

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