Red-shifted d-luciferin analogues and their bioluminescence characteristics
2
Issued Date
2025-01-01
Resource Type
eISSN
26330679
Scopus ID
2-s2.0-105023420210
Journal Title
Rsc Chemical Biology
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SCOPUS
Bibliographic Citation
Rsc Chemical Biology (2025)
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Watthaisong P., Kantiwiriyawanitch C., Jitkaroon W., Phintha A., Klayparn I., Lawan N., Kamutira P., Sasaki D., Visitsatthawong S., Maenpuen S., Tinikul R., Sucharitakul J., Nishihara R., Niwa K., Nakajima Y., Ohmiya Y., Chaiyen P. Red-shifted d-luciferin analogues and their bioluminescence characteristics. Rsc Chemical Biology (2025). doi:10.1039/d5cb00287g Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/113437
Title
Red-shifted d-luciferin analogues and their bioluminescence characteristics
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Abstract
d-Luciferin (d-LH<inf>2</inf>) is the most used substrate for beetle luciferases in various bioluminescence applications. Here, we successfully synthesized six d-LH<inf>2</inf> analogues including 5′,7′-dimethoxy-d-LH<inf>2</inf> and 7′-methylnaphthol-d-LH<inf>2</inf> as novel compounds. We also developed a continuous one-pot green synthesis method to improve yields of luciferins from condensation of quinone and d-Cys (63-fold greater than the previous report). The novel d-LH<inf>2</inf> analogues were tested with five luciferases (Fluc, SLR, Eluc, Pmluc-WT, and Pmluc-N230S), and all the compounds emitted bioluminescence at wavelengths longer than that of d-LH<inf>2</inf> (>80 nm). The reaction of SLR with 5′,7′-dimethoxy-d-LH<inf>2</inf> gave the longest red-shifted bioluminescence at 663 nm. Remarkably, the reactions of 5′-methyl-d-LH<inf>2</inf> emit longer wavelengths and brighter light than those of d-LH<inf>2</inf> in all tested luciferases, except for Eluc. Interestingly, the novel red-shifted 5′,7′-dimethyl-d-LH<inf>2</inf> also provided prolonged bioluminescence with a rate of light decay slower than that of d-LH<inf>2</inf>. We further demonstrated applications of 5′-methyl-d-LH<inf>2</inf> and 5′,7′-dimethyl-d-LH<inf>2</inf> in mammalian cell lines expressing Fluc, SLR, and Pmluc-N230S. 5′-Methyl-d-LH<inf>2</inf> provided about 11.2-fold greater sensitivity to detect Fluc in the HEK293T crude lysate than d-LH<inf>2</inf>, achieving the detection with a lower number of cell lines. The red-shifted 5′,7′-dimethyl-d-LH<inf>2</inf> also exhibits high sensitivity when using a red light filter to monitor live cell bioluminescence. These d-LH<inf>2</inf> analogues, 5′-methyl-d-LH<inf>2</inf> and 5′,7′-dimethyl-d-LH<inf>2</inf>, are promising substrates for future cell-based assays and real-time monitoring applications.