title: Latent luciferase activity in the fruit fly revealed by a synthetic luciferin.
abstract: Beetle luciferases are thought to have evolved from fatty acyl-CoA synthetases present in all insects. Both classes of enzymes activate fatty acids with ATP to form acyl-adenylate intermediates, but only luciferases can activate and oxidize d-luciferin to emit light. Here we show that the Drosophila fatty acyl-CoA synthetase CG6178, which cannot use d-luciferin as a substrate, is able to catalyze light emission from the synthetic luciferin analog CycLuc2. Bioluminescence can be detected from the purified protein, live Drosophila Schneider 2 cells, and from mammalian cells transfected with CG6178. Thus, the nonluminescent fruit fly possesses an inherent capacity for bioluminescence that is only revealed upon treatment with a xenobiotic molecule. This result expands the scope of bioluminescence and demonstrates that the introduction of a new substrate can unmask latent enzymatic activity that differs significantly from an enzyme's normal function without requiring mutation.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605.
authors.forename: David M
authors.initials: DM
authors.lastname: Mofford
authors.forename: Gadarla Randheer
authors.initials: GR
authors.lastname: Reddy
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Miller
mesh_terms: D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003412:Cricetulus; D004331:Drosophila melanogaster / Q000201:enzymology*; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism*
chemical_list: D013844:Thiazoles; D008156:Luciferases
keywords: chemical biology; enzymatic promiscuity; evolution; firefly luciferase; imaging