|
ИСТИНА |
Войти в систему Регистрация |
ИСТИНА ЦЭМИ РАН |
||
Recombinant human erythropoietin with additional processable protein domains: purification of protein synthesized in Escherichia coli heterologous expression systemстатья
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 4 января 2018 г.
- Авторы: Grunina T.M., Demidenko A.V., Lyaschuk A.M., Poponova M.S., Galushkina Z.M., Soboleva L.A., Cherepushkin S.A., Polyakov N.B., Grumov D.A., Solovyev A.I., Zhukhovitsky V.G., Boksha I.S., Subbotina M.E., Gromov A.V., Lunin V.G., Karyagina A.S.
- Журнал: Biochemistry
- Том: 82
- Номер: 11
- Год издания: 2017
- Издательство: American Chemical Society
- Местоположение издательства: United States
- Первая страница: 1285
- Последняя страница: 1294
- DOI: 10.1134/S0006297917110062
- Аннотация: Three variants of human recombinant erythropoietin (rhEPO) with additional N-terminal protein domains were obtained by synthesis in an Escherichia coli heterologous expression system. These domains included (i) maltose binding protein (MBP), (ii) MBP with six histidine residues (6His) in N-terminal position, (iii) s-tag (15 a.a. oligopeptide derived from bovine pancreatic ribonuclease A) with N-terminal 6His. Both variants of the chimeric protein containing MBP domain were prone to aggregation under nondenaturing conditions, and further purification of EPO after the domain cleavage by enterokinase proved to be impossible. In the case of 6His-s-tag-EPO chimeric protein, the products obtained after cleavage with enterokinase were successfully separated by column chromatography, and rhEPO without additional domains was obtained. Results of MALDI-TOF mass spectrometry showed that after refolding 6His-s-tag-EPO formed a structure similar to that of native EPO with two disulfide bonds. Both 6His-s-tag-EPO and rhEPO without additional protein domains purified after proteolysis possessed biological activity in vitro in the cell culture.
- Добавил в систему: Карягина-Жулина Анна Станиславовна