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Fluorescent protein with environmentally-sensitive fluorescence lifetime for quantitative pH measurementстатья
Статья опубликована в высокорейтинговом журнале
Статья опубликована в журнале из списка Web of Science и/или Scopus- Авторы: Simonyan Tatiana R., Protasova Elena A., Mamontova Anastasia V., Shakhov Aleksander M., Bodunova Daria V., Sidorenko Svetlana V., Maksimov Eugene G., Bogdanov Alexey M.
- Журнал: Archives of Biochemistry and Biophysics
- Том: 766
- Год издания: 2025
- Издательство: Academic Press
- Местоположение издательства: United States
- Номер статьи: 110350
- DOI: 10.1016/j.abb.2025.110350
- Аннотация: ntracellular pH is a key factor in cell homeostasis, regulated within specific compartments, and changes in pH can result from or affect biochemical pathways. This study explores a yellow fluorescent protein EYFP-G65T as a core for a time-resolved pH-indicator. Among the tested designs—a circular permutant, a chimeric SypHer3s-like construct, and an unmodified protein—the unmodified EYFP-G65T performed best for live-cell imaging. Upon two-photon excitation, purified EYFP-G65T exhibited a 4.5-fold increase in mean fluorescence lifetime across pH 5.5–7 and a 7-fold change in its major component's lifetime from pH 6.5–8. Using this indicator, we measured pH values ranging from 6 to 8 in various organelles, and mapped pH shifts in mitochondria and the Golgi apparatus in response to stimuli.
- Добавил в систему: Сидоренко Светлана Вадимовна