Biopolym. Cell. 2020; 36(4):254-263.
Структура та функції біополімерів
Метилування фактора елонгації трансляції людини еЕF1А2 не впливає на взаємодію еEF1A2-еEF1B.
- Інститут молекулярної біології і генетики НАН України
Вул. Академіка Заболотного, 150, Київ, Україна, 03143 - Інститут експериментальної патології, онкології і радіобіології ім. Р. Є. Кавецького
вул. Васильківська, 45, Київ, Україна, 03022
Abstract
Мета. З’ясування впливу метилування лізинових залишків фактора елонгації трансляції еЕF1А2 на взаємодію із субодиницями елонгаційного комплексу еЕF1В. Методи. Мутагенез, клонування, культивування клітин людини 293, трансфекція клітин, BRET (резонанс трансферу біолюмінісцентної енергії), афінна очистка білкових комплексів, Вестерн блот, денситометрія. Результати. Отримані п’ять мутантів еЕF1А2, в яких один з п’яти консервативних лізинових залишків, здатних до метилування, замінений на аргінін, та один мутант, в якого мутовані всі п’ять лізинів. Результати BRET та Вестерн блот аналізу виділених білків-партнерівеЕF1А2 показали відсутність суттєвих відмінностей у взаємодії еЕF1А2 і його мутантів, не здатних до метилування, із субодиницями елонгаційного комплексу еEF1B. Висновки. Метилування лізинових залишків еЕF1А2, вочевидь, не впливає на взаємодію із субодиницями елонгаційного комплексу еEF1B.
Keywords: еЕF1А2, еЕА1Вα, еЕА1Вβ, еЕА1Вγ, метилування, BRET, HaloTag.
Повний текст: (PDF, англійською)
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