A Route from Darkness to Light: Emergence and Evolution of Luciferase Activity in AMP-CoA-Ligases Inferred from a Mealworm Luciferase-like Enzyme
| Autor: | Rogilene A. Prado, Vadim R. Viviani, Dai-ichiro Kato, D. R. Neves, João Alexandre Ribeiro Gonçalves Barbosa |
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| Rok vydání: | 2013 |
| Předmět: |
Models
Molecular Oxygenase Firefly luciferin Firefly Luciferin Biology Biochemistry Evolution Molecular chemistry.chemical_compound Naphthalenesulfonates Catalytic Domain Coenzyme A Ligases Botany Animals Bioluminescence Luciferase Amino Acid Sequence Enzyme kinetics Luciferases Tenebrio Fluorescent Dyes Substrate (chemistry) Luciferin Kinetics Amino Acid Substitution chemistry Mutagenesis Site-Directed Insect Proteins Acyl Coenzyme A Protein Binding |
| Zdroj: | Biochemistry. 52:3963-3973 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi400141u |
| Popis: | The origin of luciferases and of bioluminescence is enigmatic. In beetles, luciferases seem to have evolved from AMP-CoA-ligases. How the new oxygenase luminogenic function originated from AMP-ligases leading to luciferases is one of the most challenging mysteries of bioluminescence. Comparison of the cloned luciferase-like enzyme from the nonluminescent Zophobas morio mealworm and beetle luciferases showed that the oxygenase activity may have emerged as a stereoselective oxidative drift with d-luciferin, a substrate that cannot be easily thioesterified to CoA as in the case of the l-isomer. While the overall kcat displayed by beetle luciferases is orders of magnitude greater than that of the luciferase-like enzyme, the respective oxidation rates and quantum yields of bioluminescence are roughly similar, suggesting that the rate constant of the AMP-ligase activity exerted on the new d-luciferin substrate in beetle protoluciferases was the main enzymatic property that suffered optimization during the evolution of luciferases. The luciferase-like enzyme and luciferases boost the rate of luciferyl-adenylate chemiluminescent oxidation by factors of 10(6) and 10(7), respectively, as compared to the substrate spontaneous oxidation in buffer. A similar enhancement of luciferyl-adenylate chemiluminescence is provided by nucleophilic aprotic solvents, implying that the peptide bonds in the luciferin binding site of beetle luciferase could provide a similar catalytically favorable environment. These data suggest that the luciferase-like enzyme and other similar AMP-ligases are potential alternative oxygenases. Site-directed mutagenesis studies of the luciferase-like enzyme and the red light-producing luciferase of Phrixotrix hirtus railroadworm confirm here a critical role for T/S345 in luciferase function. Mutations such as I327T/S in the luciferase-like enzyme, which simultaneously increases luciferase activity and promotes blue shifts in the emission spectrum, could have been critical for evolving functional bioluminescence from red-emitting protoluciferases. Through the combination of I327T/S mutations and N-terminal fusion, the luminescence activity of this enzyme was increased to visible levels, with the development of a totally new orange-emitting luciferase. These results open the possibility of engineering luciferase activity in a set of AMP-CoA-ligases. |
| Databáze: | OpenAIRE |
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