Illuminating the mechanism and allosteric behavior of NanoLuc luciferase

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Publikace nespadá pod Ekonomicko-správní fakultu, ale pod Přírodovědeckou fakultu. Oficiální stránka publikace je na webu muni.cz.
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NEMERGUT Michal PLUSKAL Daniel HORÁČKOVÁ Jana ŠUSTROVÁ Tereza TULIS Jan BÁRTA Tomáš BAATALLAH Racha GAGNOT Glwadys NOVÁKOVÁ Veronika MAJEROVÁ Marika SEDLÁČKOVÁ Karolina MARQUES Sérgio Manuel TOUL Martin DAMBORSKÝ Jiří PROKOP Zbyněk BEDNÁŘ David JANIN Yves L. MAREK Martin

Rok publikování 2023
Druh Článek v odborném periodiku
Časopis / Zdroj Nature Communications
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www https://www.nature.com/articles/s41467-023-43403-y
Doi http://dx.doi.org/10.1038/s41467-023-43403-y
Klíčová slova MOLECULAR-DYNAMICS SIMULATIONS; BIOLOGICAL MACROMOLECULES; CRYSTAL-STRUCTURE; BINDING PROTEIN; CDNA CLONING; BIOLUMINESCENCE; COELENTERAMIDE; INTEGRATION; PREDICTION; SUBSTRATE
Přiložené soubory
Popis NanoLuc, a superior beta-barrel fold luciferase, was engineered 10 years ago but the nature of its catalysis remains puzzling. Here experimental and computational techniques are combined, revealing that imidazopyrazinone luciferins bind to an intra-barrel catalytic site but also to an allosteric site shaped on the enzyme surface. Structurally, binding to the allosteric site prevents simultaneous binding to the catalytic site, and vice versa, through concerted conformational changes. We demonstrate that restructuration of the allosteric site can boost the luminescent reaction in the remote active site. Mechanistically, an intra-barrel arginine coordinates the imidazopyrazinone component of luciferin, which reacts with O2 via a radical charge-transfer mechanism, and then it also protonates the resulting excited amide product to form a light-emitting neutral species. Concomitantly, an aspartate, supported by two tyrosines, fine-tunes the blue color emitter to secure a high emission intensity. This information is critical to engineering the next-generation of ultrasensitive bioluminescent reporters.
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