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FruitFire: a luciferase based on a fruit fly metabolic enzyme
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 arti...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10327219/ https://www.ncbi.nlm.nih.gov/pubmed/37425765 http://dx.doi.org/10.1101/2023.06.30.547126 |
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author | Adams, Spencer T. Zephyr, Jacqueto Bohn, Markus F. Schiffer, Celia A. Miller, Stephen C. |
author_facet | Adams, Spencer T. Zephyr, Jacqueto Bohn, Markus F. Schiffer, Celia A. Miller, Stephen C. |
author_sort | Adams, Spencer T. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-10327219 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-103272192023-07-08 FruitFire: a luciferase based on a fruit fly metabolic enzyme Adams, Spencer T. Zephyr, Jacqueto Bohn, Markus F. Schiffer, Celia A. Miller, Stephen C. bioRxiv Article 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. Cold Spring Harbor Laboratory 2023-06-30 /pmc/articles/PMC10327219/ /pubmed/37425765 http://dx.doi.org/10.1101/2023.06.30.547126 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use. |
spellingShingle | Article Adams, Spencer T. Zephyr, Jacqueto Bohn, Markus F. Schiffer, Celia A. Miller, Stephen C. FruitFire: a luciferase based on a fruit fly metabolic enzyme |
title | FruitFire: a luciferase based on a fruit fly metabolic enzyme |
title_full | FruitFire: a luciferase based on a fruit fly metabolic enzyme |
title_fullStr | FruitFire: a luciferase based on a fruit fly metabolic enzyme |
title_full_unstemmed | FruitFire: a luciferase based on a fruit fly metabolic enzyme |
title_short | FruitFire: a luciferase based on a fruit fly metabolic enzyme |
title_sort | fruitfire: a luciferase based on a fruit fly metabolic enzyme |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10327219/ https://www.ncbi.nlm.nih.gov/pubmed/37425765 http://dx.doi.org/10.1101/2023.06.30.547126 |
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