Quinoline-Malononitrile-Based Aggregation-Induced Emission Probe for Monoamine Oxidase Detection in Living Cells

被引:6
作者
Duangkamol, Chuthamat [1 ,2 ]
Wangngae, Sirilak [1 ]
Wet-osot, Sirawit [3 ]
Khaikate, Onnicha [1 ]
Chansaenpak, Kantapat [4 ]
Lai, Rung-Yi [1 ]
Kamkaew, Anyanee [1 ]
机构
[1] Suranaree Univ Technol, Inst Sci, Sch Chem, Nakhon Ratchasima 30000, Thailand
[2] Pathumthani Univ, Fac Allied Hlth Sci, Div Basic & Med Sci, Pathum Thani 12000, Thailand
[3] Minist Publ Hlth, Dept Med Sci, Med Life Sci Inst, Nonthaburi 11000, Thailand
[4] Natl Nanotechnol Ctr, Natl Sci & Technol Dev Agcy, Thailand Sci Pk, Pathum Thani 12120, Thailand
关键词
AIE; activity-based fluorescence probe; MAO; enzymatic detection; AIEgen; MAO-A; CLORGYLINE; DESIGN;
D O I
10.3390/molecules28062655
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A quinoline-malononitrile (QM)-based aggregation-induced emission probe was developed to detect MAOs in cells through an enzymatic reaction followed by beta-elimination. After being incubated at 37 degrees C, QM-NH2 responded to the MAO enzymes with great specificity and within just 5 min. This 5 min responsive mechanism was fast, with the limit of detection (LOD) at 5.49 and 4.76 mu g mL(-1) for MAO-A and MAO-B, respectively. Moreover, QM-NH2 displayed high enzyme specificity even in the presence of high concentrations of biological interferences, such as oxidizing and reducing agents, biothiols, amino acids, and glucose. Furthermore, QM-NH2 demonstrated biocompatibility as the cells retained more than 70% viability when exposed to QM-NH2 at concentrations of up to 20 mu M. As a result, QM-NH2 was used to detect MAO-A and MAO-B in SH-SY5Y and HepG2 cells, respectively. After 1h incubation with QM-NH2, the cells exhibited enhanced fluorescence by about 20-fold. Moreover, the signal from cells was reduced when MAO inhibitors were applied prior to incubating with QM-NH2. Therefore, our research recommends using a QM probe as a generic method for producing recognition moieties for fluorogenic enzyme probes.
引用
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页数:14
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