Biopolym. Cell. 2020; 36(4):254-263.
Структура и функции биополимеров
Метилирование фактора элонгации трансляции человека ЕЕF1А2 не влияет на взаимодействие еEF1A2-еEF1B.
1Порублёва Л. В., 2Колесник Д. Л., 1Ельская А. В., 1Негруцкий Б. С.
  1. Институт молекулярной биологии и генетики НАН Украины
    ул. Академика Заболотного, 150, Киев, Украина, 03143
  2. Институт экспериментальной патологии, онкологии и радиобиологии им. Р. Е. Кавецкого
    ул. Васильковская, 45, Киев, Украина, 03022

Abstract

Цель. Выяснение влияния метилирования лизиновых остатков фактора элонгации трансляции еЕF1А2 на взаимодействие с субъединицами элонгационного комплекса еЕF1В. Методы. Мутагенез, клонирование, культивирование клеток человека 293, трансфекция клеток, BRET (резонанс трансфера биолюминисцентной энергии), аффинная очистка белковых комплексов, Вестерн блот, денситометрия. Результаты. Получены пять мутантов еЕF1А2, в которых один из пяти метилируемых консервативных лизиновых остатков, заменен на аргинин, и один мутант, у которого мутированы все пять лизинов. Результаты BRET и Вестерн блот анализа выделенных белков-партнеров еЕF1А2 не показали существенных различий во взаимодействии еЕF1А2 и его неметилируемых мутантов с субъединицами элонгационного комплексаеЕF1В. Выводы. Метилирование лизиновых остатков еЕF1А2, по-видимому, не влияет на взаимодействие с субъединицами элонгационного комплекса еЕF1В.
Keywords: еЕF1А2, еЕА1Вα, еЕА1Вβ, еЕА1Вγ, метилирование, BRET, HaloTag.

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