Reaction-Based Genetically Encoded Fluorescent Hydrogen Sulfide Sensors
Reaction-Based Genetically Encoded Fluorescent Hydrogen Sulfide Sensors
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DOI：
10.1021/ja303261d
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发表时间：
2012-06-13
影响因子：
15
通讯作者：
Ai, Hui-wang
中科院分区：
文献类型：
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作者：
Chen, Si;Chen, Zhi-jie;Ai, Hui-wang
硫化氢（H2S）是一种有毒气体，也是一种重要的生物信号分子，其检测一直是一个长期的挑战。在这里，我们报告了基于基因编码的荧光蛋白（FP）的探针，可以选择性地检测 H2S。通过扩展大肠杆菌和哺乳动物细胞的遗传密码，FP 发色团被硫醚反应性叠氮官能团修饰。这些结构修饰的发色团被 H2S 选择性还原，从而产生可通过光谱和显微技术检测到的灵敏荧光增强。对循环排列 FP 的探索导致了响应速度更快的改进传感器，并且使用这种基因编码探针监测活哺乳动物细胞中 H2S 的可行性也已得到证明。
The detection of hydrogen sulfide (H2S), a toxic gas and an important biological signaling molecule, has been a long-time challenge. Here we report genetically encoded fluorescent protein (FP)-based probes that can selectively detect H2S. By expanding the genetic codes of E. coli and mammalian cells, FP chromophores were modified with the sulfide-reactive azide functional group. These structurally modified chromophores were selectively reduced by H2S, resulting in sensitive fluorescence enhancement detectable by spectroscopic and microscopic techniques. Exploration of a circularly permuted FP led to an improved sensor with faster responses, and the feasibility of using such a genetically encoded probe to monitor H2S in living mammalian cells has also been demonstrated.