The nuclear factor κB inhibitor (E)-2-fluoro-4'-methoxystilbene inhibits firefly luciferase.
Photinus pyralis (firefly) luciferase is widely used as a reporter system to monitor alterations in gene promoter and/or signalling pathway activities in vitro. The enzyme catalyses the formation of oxyluciferin from D-luciferin in an ATP-consuming reaction involving photon emission. The purpose of the present study was to characterize the luciferase-inhibiting potential of (E)-2-fluoro-4'-methoxystilbene, which is known as a potent inhibitor of the NF-κB (nuclear factor κB) signalling pathway that is used to modulate the NF-κB signalling pathway in vitro. Results show that (E)-2-fluoro-4'-methoxystilbene effectively inhibits firefly luciferase activity in cell lysates and living cells in a non-competitive manner with respect to the luciferase substrates D-luciferin and ATP. By contrast, the compound has no effect on Renilla and Gaussia luciferases. The mechanism of firefly luciferase inhibition by (E)-2-fluoro-4'-methoxystilbene, as well as its potency is comparable to its structure analogue resveratrol. The in vitro use of trans-stilbenes such as (E)-2-fluoro-4'-methoxystilbene or resveratrol compromises firefly luciferase reporter assays as well as ATP/luciferase-based cell viability assays.
Department of Toxicology, Institute of Experimental and Clinical Pharmacology and Toxicology, University of Tübingen, Tübingen, Germany. albert.braeuning@uni-tuebingen.de
Albert
A
Braeuning
Silvia
S
Vetter
D000818:Animals; D045744:Cell Line, Tumor; D033342:Copepoda / Q000201:enzymology; D048888:Fireflies / Q000201:enzymology*; D020128:Inhibitory Concentration 50; D008156:Luciferases / Q000037:antagonists & inhibitors / Q000378:metabolism; D049409:Luciferases, Firefly / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D049410:Luciferases, Renilla / Q000037:antagonists & inhibitors / Q000378:metabolism; D049408:Luminescent Agents / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D048790:Renilla / Q000201:enzymology; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*
C569653:2-fluoro-4'-methoxystilbene; D049408:Luminescent Agents; D016328:NF-kappa B; D013267:Stilbenes; D008156:Luciferases; D049410:Luciferases, Renilla; D049409:Luciferases, Firefly; D000077185:Resveratrol