Genetically Encoded Fluorescent Probe for the Selective Detection of Peroxynitrite
Genetically Encoded Fluorescent Probe for the Selective Detection of Peroxynitrite
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DOI：
10.1021/ja408011q
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发表时间：
2013-10-09
影响因子：
15
通讯作者：
Ai, Hui-wang
中科院分区：
文献类型：
--
作者：
Chen, Zhi-jie;Ren, Wei;Ai, Hui-wang
过氧亚硝酸盐是参与细胞信号传导和病理过程的高反应性分子。我们在此报告了一种新的基因编码探针，pnGFP，它可以选择性地感知过氧亚硝酸根。将硼酸部分位点特异性地引入到环状排列的荧光蛋白中。通过研究不同的蛋白质模板，然后定点随机诱变，我们确定了一个选择性过氧亚硝酸盐传感器，这是基本上不响应其他常见的细胞氧化还原信号分子。这种新的探针已被遗传引入哺乳动物细胞中，以在生理相关浓度下对过氧亚硝酸盐进行成像。
Peroxynitrite is a highly reactive molecule involved in cell signaling and pathological processes. We hereby report a novel genetically encoded probe, pnGFP, which can selectively sense peroxynitrite. A boronic acid moiety was site-specifically introduced into circularly permuted fluorescent proteins. By examining different protein templates followed with site-targeted random mutagenesis, we identified a selective peroxynitrite sensor, which is essentially unresponsive to other common cellular redox signaling molecules. The new probe has been genetically introduced into mammalian cells to image peroxynitrite at physiologically relevant concentrations.