通过卤代标签蛋白标记对活细胞中的细胞器特异性一氧化氮进行检测
Organelle-Specific Nitric Oxide Detection in Living Cells via HaloTag Protein Labeling.
作者信息
Wang Jianhua, Zhao Yuzheng, Wang Chao, Zhu Qian, Du Zengmin, Hu Aiguo, Yang Yi
机构信息
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, CAS Center for Excellence in Brain Science, School of Pharmacy, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
Shanghai Key Laboratory of Advanced Polymeric Materials, School of Materials Science and Engineering, East China University of Science and Technology, 130 Mei Long Road, Shanghai, 200237, P. R. China.
出版信息
PLoS One. 2015 Apr 29;10(4):e0123986. doi: 10.1371/journal.pone.0123986. eCollection 2015.
Nitric oxide (NO) is a membrane-permeable signaling molecule that is constantly produced, transferred, and consumed in vivo. NO participates and plays important roles in multiple biological processes. However, spatiotemporal imaging of NO in living cells is challenging. To fill the gap in currently used techniques, we exploited the versatility of HaloTag technology and synthesized a novel organelle-targetable fluorescent probe called HTDAF-2DA. We demonstrate the utility of the probe by monitoring subcellular NO dynamics. The developed strategy enables precise determination of local NO function.
一氧化氮（NO）是一种可透过细胞膜的信号分子，在体内不断产生、传递和消耗。NO参与多种生物学过程并发挥重要作用。然而，对活细胞中NO进行时空成像具有挑战性。为了填补当前所用技术的空白，我们利用了HaloTag技术的多功能性，合成了一种名为HTDAF-2DA的新型细胞器靶向荧光探针。我们通过监测亚细胞水平的NO动态来证明该探针的实用性。所开发的策略能够精确确定局部NO的功能。
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