Jia Zhiyuan, Han Hai-Hao, Sedgwick Adam C, Williams George T, Gwynne Lauren, Brewster James T, Bull Steven D, Jenkins A Toby A, He Xiao-Peng, Schönherr Holger, Sessler Jonathan L, James Tony D
Department of Chemistry and Biology, Physical Chemistry & Research Center of Micro- and Nanochemistry and Engineering (Cμ), University of Siegen, Siegen, Germany.
Key Laboratory for Advanced Materials and Joint International Research Laboratory of Precision Chemistry and Molecular Engineering, Feringa Nobel Prize Scientist Joint Research Center, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai, China.
Front Chem. 2020 Jun 3;8:389. doi: 10.3389/fchem.2020.00389. eCollection 2020.
Here, we report a new pentafluoropropanamido rhodamine fluorescent probe (ACS-HNE) that allows for the selective detection of neutrophil elastase (NE). ACS-HNE displayed high sensitivity, with a low limit of detection (<5.3 nM), and excellent selectivity toward elastase over other relevant biological analytes and enzymes. The comparatively poor solubility and cell permeability of neat ACS-HNE was improved by creating an ACS-HNE-albumin complex; this approach allowed for improvements in the visualization of elastase activity in RAW 264.7 cells relative to ACS-HNE alone. The present study thus serves to demonstrate a simple universal strategy that may be used to overcome cell impermeability and solubility limitations, and to prepare probes suitable for the cellular imaging of enzymatic activity .
在此，我们报道了一种新型五氟丙酰胺罗丹明荧光探针（ACS-HNE），它能够选择性检测中性粒细胞弹性蛋白酶（NE）。ACS-HNE表现出高灵敏度，检测限低（<5.3 nM），并且相对于其他相关生物分析物和酶，对弹性蛋白酶具有出色的选择性。通过形成ACS-HNE-白蛋白复合物，改善了纯ACS-HNE相对较差的溶解性和细胞通透性；相对于单独的ACS-HNE，这种方法提高了RAW 264.7细胞中弹性蛋白酶活性的可视化效果。因此，本研究旨在证明一种简单通用的策略，该策略可用于克服细胞不透性和溶解性限制，并制备适用于酶活性细胞成像的探针。
