Recombinant alpha-fetoprotein receptor-binding domain co-expression with polyglutamate tags facilitates in vivo folding in E. coli.

Mollaev M., Gorokhovets N., Nikolskaya E., Faustova M., Zabolotsky A., Sokol M., Tereshenko O., Zhunina O., Shvets V., Severin E., Yabbarov N.
Тип документаJournal Article
Дата публикации2018-03-25
Название журналаProtein Expression and Purification
Квартиль по SCImagoQ3
Квартиль по Web of ScienceQ4
Импакт-фактор 20212.03
ISSN10465928, 10960279
Краткое описание
A wide range of methods are known to increase the prokaryotic intracellular recombinant proteins solubility, for instance, growth at low temperature, supplementation of culture media with "chemical chaperones" (proline, glycine-betaine, and trehalose), co-expression with chaperones or highly soluble fusion partners. As an alternative, we have introduced the polyglutamate tag, which, as it has been shown, increased the protein solubility and facilitated folding. In this study we evaluated the minimal quantity of high density negatively charged EEEEVE amino acid repeats (pGlu) necessary to switch the recombinant receptor-binding domain of human alpha-fetoprotein (rbdAFP) expression almost entirely from the inclusion bodies to the soluble cytoplasmic fraction in E. coli. For this purpose, genetic constructs based on pET vectors coding rbdAFP and containing from 1 to 4 additional EEEEVE repeats at the C-terminus have been prepared. It was found that 3 pGlu repeats is the minimal number, that leads to a complete shift of the expression to the soluble cytoplasmic fraction in E. coli SHuffle Express T7 while 4 repeats were required for that in E. coli BL21(DE3). The rbdAFP contained 4 pGlu repeats was purified making use of ion-exchange chromatography and characterized by circular dichroism and ability to bind and accumulate in AFP receptor positive cancer cells in order to check for the structural and specific activity alterations related to the additional polyanionic sequence introduction.
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1. Mollaev M. и др. Recombinant alpha-fetoprotein receptor-binding domain co-expression with polyglutamate tags facilitates in vivo folding in E. coli // Protein Expression and Purification. 2018. Т. 143. С. 77–82.


DO - 10.1016/j.pep.2017.11.001

UR -

TI - Recombinant alpha-fetoprotein receptor-binding domain co-expression with polyglutamate tags facilitates in vivo folding in E. coli

T2 - Protein Expression and Purification

AU - Mollaev, Murad

AU - Gorokhovets, Neonila

AU - Nikolskaya, Elena

AU - Faustova, Maria

AU - Zabolotsky, Arthur

AU - Sokol, Maria

AU - Tereshenko, Oksana

AU - Zhunina, Olga

AU - Shvets, Vitaliy

AU - Severin, Evgeniy

AU - Yabbarov, Nikita

PY - 2018

DA - 2018/03

PB - Elsevier BV

SP - 77-82

VL - 143

SN - 1046-5928

ER -

BibTex |


doi = {10.1016/j.pep.2017.11.001},

url = {},

year = 2018,

month = {mar},

publisher = {Elsevier {BV}},

volume = {143},

pages = {77--82},

author = {Murad Mollaev and Neonila Gorokhovets and Elena Nikolskaya and Maria Faustova and Arthur Zabolotsky and Maria Sokol and Oksana Tereshenko and Olga Zhunina and Vitaliy Shvets and Evgeniy Severin and Nikita Yabbarov},

title = {Recombinant alpha-fetoprotein receptor-binding domain co-expression with polyglutamate tags facilitates in vivo folding in E. coli},

journal = {Protein Expression and Purification}


Mollaev, Murad, et al. “Recombinant Alpha-Fetoprotein Receptor-Binding Domain Co-Expression with Polyglutamate Tags Facilitates in Vivo Folding in E. Coli.” Protein Expression and Purification, vol. 143, Mar. 2018, pp. 77–82. Crossref,