Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Amino Acid Substitution
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Animals
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Coenzyme A Ligases / chemistry
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Coenzyme A Ligases / genetics*
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Coenzyme A Ligases / metabolism*
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Coleoptera / enzymology
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Coleoptera / genetics
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Drosophila Proteins / chemistry
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Drosophila Proteins / genetics
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Drosophila Proteins / metabolism
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Drosophila melanogaster / enzymology
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Drosophila melanogaster / genetics
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Firefly Luciferin / metabolism
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Hydrogen Bonding
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In Vitro Techniques
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Insect Proteins / chemistry
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Insect Proteins / genetics
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Insect Proteins / metabolism
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Luciferases, Firefly / chemistry
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Luciferases, Firefly / genetics*
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Luciferases, Firefly / metabolism*
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Luminescence
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Models, Molecular
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Mutagenesis, Site-Directed
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Phylogeny
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Point Mutation
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Sequence Homology, Amino Acid
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Serine / chemistry
Substances
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Drosophila Proteins
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Insect Proteins
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Recombinant Proteins
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Serine
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Firefly Luciferin
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Luciferases, Firefly
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Coenzyme A Ligases