Firefly luciferase (Photinus-luciferin:oxygen 4-oxidoreductase) (FLuc)

Firefly luciferase is a 62 kDa bioluminescent enzyme derived from the firefly Photinus pyralis that catalyzes the oxidation of D-luciferin in the presence of ATP, magnesium, and oxygen to produce light [UniProt P08659]. In the field of drug discovery, it is extensively used as a reporter protein to study gene regulation, protein-protein interactions, and cell viability [PubMed: 18430551]. The ATP-binding site is a critical functional domain that has become a focal point for identifying assay interference; many small molecules can bind this site, acting as competitive inhibitors [PubMed: 19233613]. Such interactions can lead to false-positive results in high-throughput screening (HTS) because the inhibitors may stabilize the enzyme against degradation, leading to an increase in bioluminescent signal in certain assay formats [PubMed: 20405933]. Consequently, while not a target for treating human disease, the firefly luciferase ATP-binding site is a major off-target that must be accounted for to ensure the validity of experimental data in pharmacology [PubMed: 21114280].

Other names

Luciferin 4-monooxygenasePhotinus pyralis luciferaseATP-dependent luciferaseFirefly luciferase ATP-binding site

Competitive inhibition of the ATP-binding site