Center for Catalytic Hydrocarbon Functionalizations, Institute for Basic Science (IBS) , 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Department of Chemistry, Korea Advanced Institute of Science and Technology (KAIST) , 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
Anal Chem. 2018 Feb 20;90(4):2648-2654. doi: 10.1021/acs.analchem.7b04375. Epub 2018 Feb 6.
The development of novel fluorescent probes for monitoring the concentration of various biomolecules in living systems has great potential for eventual early diagnosis and disease intervention. Selective detection of competitive species in biological systems is a great challenge for the design and development of fluorescent probes. To improve on the design of fluorescent coumarin-based biothiol sensing technologies, we have developed herein an enhanced dual emission doubly activated system (DACP-1 and the closely related DACP-2) for the selective detection of glutathione (GSH) through the use of one optical channel and the detection of cysteine (Cys) by another channel. A phenylselenium group present at the 4-position completely quenches the fluorescence of the probe via photoinduced electron transfer to give a nonfluorescent species. Probes are selective for glutathione (GSH) in the red region and for cysteine/homocysteine (Cys/Hcy) in the green region. When they were treated with GSH, DACP-1 and DACP-2 showed strong fluorescence enhancement in comparison to that for closely related species such as amino acids, including Cys/Hcy. Fluorescence quantum yields (Φ) increased for the red channel (<0.001 to 0.52 (DACP-1) and 0.48 (DACP-2)) and green channel (Cys) (<0.001 to 0.030 (DACP-1) and 0.026 (DACP-2)), respectively. Competing fluorescent enhancements upon addition of closely related species were negligible. Fast responses, improved water solubility, and good cell membrane permeability were all properly established with the use of DACP-1 and DACP-2. Live human lung cancer cells and fibroblasts imaged by confocal microscopy, as well as live mice tumor model imaging, confirmed selective detection.
开发用于监测活系统中各种生物分子浓度的新型荧光探针，对于最终实现早期诊断和疾病干预具有很大的潜力。在生物系统中选择性地检测竞争物种是设计和开发荧光探针的一大挑战。为了改进基于荧光香豆素的生物硫醇传感技术的设计，我们在此开发了一种增强的双重发射双激活系统（DACP-1 和密切相关的 DACP-2），用于通过使用一个光学通道选择性地检测谷胱甘肽（GSH），并通过另一个通道检测半胱氨酸（Cys）。在 4 位存在的苯硒基通过光诱导电子转移完全猝灭探针的荧光，给出非荧光物质。探针对谷胱甘肽（GSH）在红色区域具有选择性，对半胱氨酸/同型半胱氨酸（Cys/Hcy）在绿色区域具有选择性。当用 GSH 处理时，DACP-1 和 DACP-2 与包括 Cys/Hcy 在内的密切相关的物质相比，显示出强荧光增强。红色通道（<0.001 至 0.52（DACP-1）和 0.48（DACP-2））和绿色通道（Cys）（<0.001 至 0.030（DACP-1）和 0.026（DACP-2））的荧光量子产率（Φ）分别增加。加入密切相关的物质后，竞争荧光增强可忽略不计。快速响应、提高的水溶性和良好的细胞膜通透性均通过 DACP-1 和 DACP-2 得到适当确立。共聚焦显微镜对活的人肺癌细胞和成纤维细胞以及活的小鼠肿瘤模型成像的确认选择性检测。
