Jiao Shan, Yang Si, Meng Xiuping, Wang Chengkun
Hospital of Stomatology, Jilin University, Qinghua Road 1500, Changchun 130021, China.
Department of Pediatric Neurology, The First Hospital of Jilin University, Xinmin Street 71, Changchun 130021, China.
Spectrochim Acta A Mol Biomol Spectrosc. 2020 Nov 5;241:118637. doi: 10.1016/j.saa.2020.118637. Epub 2020 Jun 23.
Nitroreductase (NTR) belongs to a class of flavin mononucleotide-dependent and flavin adenine dinucleotide-dependent cytoplasmic enzymes; its contents in tumor cells increase during hypoxia. The development of fluorescent probes for detection of NTR activity is of great significance for the study of the state of hypoxia in living organisms. In this paper, a red-emitting fluorescence turn-on probe EBI-NO was synthesized using a one-step method. The fluorescence of the probe was enhanced by 60 folds in the presence of NTR. The probe also had high selectivity towards NTR, and its detection limit was as low as 1 ng/mL. The reaction mechanism was verified using MS, molecular docking and theoretical calculations. In addition, it was successfully applied in real-time monitoring of NTR produced during growth of Escherichia coli (BL21) and in visualization of NTR in oral cancer cells (Cal-27) under hypoxia. This work provides a new imaging tool that can be applied to investigate the physiological and pathological changes in hypoxia oral cells.
硝基还原酶（NTR）属于一类黄素单核苷酸依赖性和黄素腺嘌呤二核苷酸依赖性细胞质酶；其在肿瘤细胞中的含量在缺氧时会增加。开发用于检测NTR活性的荧光探针对于研究生物体内的缺氧状态具有重要意义。本文采用一步法合成了一种发红光的荧光开启探针EBI-NO。在NTR存在下，该探针的荧光增强了60倍。该探针对NTR也具有高选择性，其检测限低至1 ng/mL。通过质谱、分子对接和理论计算验证了反应机理。此外，它成功应用于实时监测大肠杆菌（BL21）生长过程中产生的NTR，并用于可视化缺氧条件下口腔癌细胞（Cal-27）中的NTR。这项工作提供了一种新的成像工具，可用于研究缺氧口腔细胞中的生理和病理变化。
