Department of Chemistry, University of California, Irvine, California 92697, United States.
Molecular Biology & Biochemistry, University of California, Irvine, California 92697, United States.
J Am Chem Soc. 2022 May 4;144(17):7871-7880. doi: 10.1021/jacs.2c02058. Epub 2022 Apr 20.
Fluorogenic bioorthogonal reactions enable biomolecule visualization in real time. These reactions comprise reporters that "light up" upon reaction with complementary partners. While the spectrum of fluorogenic chemistries is expanding, few transformations are compatible with live cells due to cross-reactivities or insufficient signal turn-on. To address the need for more suitable chemistries for cellular imaging, we developed a fluorogenic reaction featuring cyclopropenone reporters and phosphines. The transformation involves regioselective activation and cyclization of cyclopropenones to form coumarin products. With optimal probes, the reaction provides >1600-fold signal turn-on, one of the highest fluorescence enhancements reported to date. The bioorthogonal motifs were evaluated in vitro and in cells. The reaction was also found to be compatible with other common fluorogenic transformations, enabling multicomponent, real-time imaging. Collectively, these data suggest that the cyclopropenone-phosphine reaction will bolster efforts to track biomolecule targets in their native settings.
荧光生物正交反应可实现生物分子的实时可视化。这些反应包含报告分子，它们与互补伴侣反应时会“点亮”。虽然荧光化学的范围正在扩大，但由于交叉反应性或信号开启不足，很少有转化适用于活细胞。为了解决更适合细胞成像的化学物质的需求，我们开发了一种具有环丙烯酮报告基团和膦的荧光生物正交反应。该转化涉及环丙烯酮的区域选择性激活和环化，形成香豆素产物。使用最佳探针，该反应提供了>1600 倍的信号开启，是迄今为止报道的最高荧光增强之一。生物正交基序在体外和细胞中进行了评估。还发现该反应与其他常见的荧光转化兼容，可实现多组分实时成像。总的来说，这些数据表明，环丙烯酮-膦反应将有助于在其天然环境中追踪生物分子靶标。
