Functional conversion of fatty acyl-CoA synthetase to firefly luciferase by site-directed mutagenesis: a key substitution responsible for luminescence activity.
We demonstrated that firefly luciferase has a catalytic function of fatty acyl-CoA synthesis [Oba, Y., Ojika, M. and Inouye, S. (2003) Firefly luciferase is a bifunctional enzyme: ATP-dependent monooxygenase and a long chain fatty acyl-CoA synthetase. FEBS Lett. 540, 251-254] and proposed that the evolutionary origin of beetle luciferase is a fatty acyl-CoA synthetase (FACS) in insect. In this study, we performed the functional conversion of FACS to luciferase by replacing a single amino acid to serine. This serine residue is conserved in luciferases and possibly interacts with luciferin. The mutants of FACSs in non-luminous click beetle Agrypnus binodulus (AbLL) and Drosophila melanogaster (CG6178) gave luminescence enhancement, suggesting that the serine residue is a key substitution responsible for luminescence activity.
Oba
Yuichi
Y
Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan. oba@agr.nagoya-u.ac.jp
Iida
Koichiro
K
Inouye
Satoshi
S
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D003066:Coenzyme A Ligases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000201:enzymology / Q000235:genetics; D049454:Firefly Luciferin / Q000378:metabolism; D006860:Hydrogen Bonding; D066298:In Vitro Techniques; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D049409:Luciferases, Firefly / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D049449:Luminescence; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010802:Phylogeny; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000737:chemistry
D029721:Drosophila Proteins; D019476:Insect Proteins; D011994:Recombinant Proteins; D012694:Serine; D049454:Firefly Luciferin; D049409:Luciferases, Firefly; D003066:Coenzyme A Ligases