БИОХИМИЯ, 2019, том 84, вып. 8, с. 1177–1188
УДК 577.6
Термостабильная лихеназа Clostridium thermocellum допускает одиночные и тандемные инсерции белковых доменов*
1 Институт физиологии растений им. К.А. Тимирязева РАН, 127276 Москва, Россия; электронная почта: Helliga.p@gmail.com и irengold58@gmail.com
2 Бакинский государственный университет, AZ 1148 Баку, Азербайджан; электронная почта: orkhan@bioset.org
Поступила в редакцию 03.03.2019
После доработки 29.04.2019
Принята к публикации 30.04.2019
DOI: 10.1134/S0320972519080104
КЛЮЧЕВЫЕ СЛОВА: лихеназа, флуоресцентные белки, одиночные и тандемные инсерции белков.
Аннотация
В этом исследовании лихеназа Clostridium thermocellum (эндо-β-1,3;1,4-глюкан-D-гликозилгидролаза, ЕС 3.2.1.73 (Р29716)) апробирована в роли белка, принимающего в двух областях инсерцию двух модельных флуоресцентных белков (EGFP и TagRFP). Функциональная укладка всех доменов подтверждена сохранением ферментативной активности лихеназы и флуоресценции EGFP и TagRFP. Полученные результаты убедительно свидетельствуют, что (i) два экспериментально апробированных петлевых региона лихеназы могут служить областями для инсерции белков без нарушения ее фолдинга in vivo, (ii) лихеназа способна допускать не только одиночные, но и тандемные вставки крупных белковых доменов. Высокая активность и термостабильность, эффективный рефолдинг in vitro лихеназы делает ее привлекательной в качестве нового принимающего «host»-белка при использовании подхода инсерционного слияния доменов в инженерии многофункциональных белков.
Текст статьи
Сноски
* Приложение к статье на английском языке опубликовано на сайте журнала «Biochemistry» (Moscow) (http://protein.bio.msu.ru/biokhimiya/) и на сайте издательства Springer (https://link.springer.com/journal/10541), том 84, вып. 8, 2019.
** Адресат для корреспонденции.
Финансирование
Работа поддержана Российским фондом фундаментальных исследований (проект № 16-34-00249; AAT).
Конфликт интересов
Авторы заявляют об отсутствии конфликта интересов.
Соблюдение этических норм
Настоящая статья не содержит описания каких-либо исследований с участием людей и животных в качестве объектов.
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