FruitFire: a luciferase based on a fruit fly metabolic enzyme.
| Autor: | Adams ST Jr; Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School, 364 Plantation St, Worcester, MA 01605., Zephyr J; Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School, 364 Plantation St, Worcester, MA 01605., Bohn MF; Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School, 364 Plantation St, Worcester, MA 01605., Schiffer CA; Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School, 364 Plantation St, Worcester, MA 01605., Miller SC; Department of Biochemistry and Molecular Biotechnology, UMass Chan Medical School, 364 Plantation St, Worcester, MA 01605. |
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| Jazyk: | angličtina |
| Zdroj: | BioRxiv : the preprint server for biology [bioRxiv] 2023 Jun 30. Date of Electronic Publication: 2023 Jun 30. |
| DOI: | 10.1101/2023.06.30.547126 |
| Abstrakt: | Firefly luciferase is homologous to fatty acyl-CoA synthetases from insects that are not bioluminescent. Here, we determined the crystal structure of the fruit fly fatty acyl-CoA synthetase CG6178 to 2.5 Å. Based on this structure, we mutated a steric protrusion in the active site to create the artificial luciferase FruitFire, which prefers the synthetic luciferin CycLuc2 to d-luciferin by >1000-fold. FruitFire enabled in vivo bioluminescence imaging in the brains of mice using the pro-luciferin CycLuc2-amide. The conversion of a fruit fly enzyme into a luciferase capable of in vivo imaging underscores the potential for bioluminescence with a range of adenylating enzymes from nonluminescent organisms, and the possibilities for application-focused design of enzyme-substrate pairs. |
| Databáze: | MEDLINE |
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