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Stalling chromophore synthesis of the fluorescent protein Venus reveals the molecular basis of the final oxidation stepстатья Исследовательская статья
Статья опубликована в высокорейтинговом журнале
Статья опубликована в журнале из списка Web of Science и/или Scopus- Авторы: Auhim Husam Sabah, Grigorenko Bella L., Harris Tessa K., Aksakal Ozan E., Polyakov Igor V., Berry Colin, Gomes Gabriel dos Passos, Alabugin Igor V., Rizkallah Pierre J., Nemukhin Alexander V., Jones D.Dafydd
- Журнал: Chemical science
- Год издания: 2021
- Издательство: Royal Society of Chemistry
- Местоположение издательства: United Kingdom
- DOI: 10.1039/d0sc06693a
- Аннотация: Fluorescent proteins (FPs) have revolutionised the life sciences, but the mechanism of chromophorematuration is still not fully understood. Here we show that incorporation of a photo-responsive noncanonical amino acid within the chromophore stalls maturation of Venus, a yellow FP, at an intermediate stage; a crystal structure indicates the presence of O2 located above a dehydrated enolate form of the imidazolone ring, close to the strictly conserved Gly67 that occupies a twisted conformation. His148 adopts an “open” conformation so forming a channel that allows O2 access to the immature chromophore. Absorbance spectroscopy supported by QM/MM simulations suggests that the first oxidation step involves formation of a hydroperoxyl intermediate in conjunction with dehydrogenation of the methylene bridge. A fully conjugated mature chromophore is formed through release of H2O2, both in vitro and in vivo. The possibility of interrupting and photochemically restarting chromophore maturation and the mechanistic insights open up new approaches for engineering optically controlled fluorescent proteins.
- Добавил в систему: Немухин Александр Владимирович
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