Sensing cytochrome P450 1A1 activity by a resorufin-based isoform-specific fluorescent probe |
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| Affiliation: | 1. Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;2. College of Pharmacy, Dalian Medical University, Dalian 116044, China;1. College of Chemistry and Chemical Engineering, Yunnan Normal University, Kunming 650500, China;2. College of Chemical Science and Technology, Yunnan University, Kunming 650091, China;1. Department of Anatomy, Faculty of Medicine, P.J. Šafárik University, Šrobárova 2, 041 80 Košice, Slovak Republic;2. Department of Pharmacology, Faculty of Medicine, P.J. Šafárik University, tr. SNP 1, 040 11 Košice, Slovak Republic;1. Department of Biomedical Science, The University of Nottingham Malaysia Campus, Jalan Broga, 43500 Semenyih, Selangor Darul Ehsan, Malaysia;2. School of Medical Sciences, International Medical University, 126, Jalan 19/155B, Bukit Jalil, 57000 Kuala Lumpur, Malaysia;3. Herbal Medicine Research Unit, Division of Biochemistry, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia;4. Centre of Excellence for Research in AIDS (CERiA), Universiti Malaya, Level 17 Wisma R&D, Jalan Pantai Baru, 59990 Kuala Lumpur, Malaysia;5. School of Pharmacy, Monash University Malaysia, Jalan Lagoon Selatan, 47500 Bandar Sunway, Selangor Darul Ehsan, Malaysia;1. Faculty of Engineering and Information Technology, Taiz University, Taiz, 6803, Yemen;2. School of Engineering Sciences and Technology (SEST), University of Hyderabad, Hyderabad, 500046, India;3. Department of Chemistry, Faculty of Science, King Khalid University, Abha, Saudi Arabia;4. Energy & Resource Management Division, CSIR-National Environmental Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur, 440 020, India;5. Department of Physics, Faculty of Science, Al-Azhar University, Nasr City, 11884, Cairo, Egypt |
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| Abstract: | Cytochrome P450 1A1 (CYP1A1), a heme-containing monooxygenase, is of particular importance for human health because of its vital roles in the metabolic activation of pro-carcinogenic compounds to the carcinogens. Deciphering the relevance of CYP1A1 to human diseases and screening of CYP1A1 modulators require reliable tool(s) for probing this key enzyme in complex biological matrices. Herein, a practical and ultrasensitive fluorescence-based assay for real-time sensing CYP1A1 activities in biological systems has been developed, via designing an isoform-specific fluorogenic sensor for CYP1A1 (CHPO). The newly developed fluorogenic substrate for CYP1A1 has been carefully investigated in terms of specificity, sensitivity, precision, quantitative linear range and the anti-interference ability. The excellent selectivity, strong anti-interference ability and fast response kinetics, making the practicability of CHPO-based CYP1A1 activity assay is better than that of most reported CYP1A1 activity assays. Furthermore, CHPO has been successfully used for imaging CYP1A1 activities in living cells and human tissues, as well as for high-throughput screening of CYP1A1 inhibitors using tissue preparations as enzyme sources. Collectively, this study provided a practical fluorogenic sensor for real-time sensing CYP1A1 in complex biological systems, which would strongly facilitate the investigations on the relevance of CYP1A1 to human diseases and promote high-throughput screening of CYP1A1 modulators for biomedical applications. |
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| Keywords: | Cytochrome P450 1A1 Fluorogenic sensor Activity sensing Resorufin Isoform-specificity Bioimaging |
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