Evolution of fluorescein as a platform for finely tunable fluorescence probes
Evolution of fluorescein as a platform for finely tunable fluorescence probes
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DOI：
10.1021/ja043919h
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发表时间：
2005-04-06
影响因子：
15
通讯作者：
Nagano, T
中科院分区：
文献类型：
--
作者：
Urano, Y;Kamiya, M;Nagano, T
荧光成像是目前可用于连续观察活细胞内动态细胞内过程的最强大的技术。合适的荧光探针对于荧光成像自然是至关重要的，但由于缺乏灵活的荧光探针设计策略，目前只能显示非常有限的生物分子。目前，设计在很大程度上是经验的。在这里，我们展示了传统荧光素染料的羧基，以前被认为是必不可少的，已经被其他取代基取代，提供了各种新的荧光素。此外，通过突破荧光素的传统结构，我们在严格的光化学基础上开发出了第一个完全合理的新型荧光探针的设计策略。这种方法的价值体现在它应用于开发一种新型的、高灵敏度的、膜透过性的荧光探针来检测最广泛使用的报告酶--β-半乳糖苷酶。
Fluorescence imaging is the most powerful technique currently available for continuous observation of dynamic intracellular processes in living cells. Suitable fluorescence probes are naturally of critical importance for fluorescence imaging, but only a very limited range of biomolecules can currently be visualized because of the lack of flexible design strategies for fluorescence probes. At present, design is largely empirical. Here we show that the carboxylic group of traditional fluorescein dyes, formerly considered indispensable, has been replaced with other substituents, affording various kinds of new fluoresceins. Further, by breaking out of the traditional structure of fluorescein, we developed the first and totally rational design strategy for novel fluorescence probes based on a strict photochemical basis. The value of this approach is exemplified by its application to develop a novel, highly sensitive, and membrane-permeable fluorescence probe for beta-galactosidase, which is the most widely used reporter enzyme.