Human IL-36RA production in Escherichia coli with coexpression of E. coli methionine aminopeptidase. II. Comparison of IL-36RA biological activity from different strains
- Authors: Kolobov A.A.1,2, Kondratyeva E.V.1, Sharafutdinova T.A.1, Kalinin R.S.1,2, Nimiritsky P.P.2, Stefanov V.E.2, Petrov A.V.1
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Affiliations:
- Institute of Highly Pure Biopreparations
- Department of Biochemistry
- Issue: Vol 11, No 6 (2017)
- Pages: 453-457
- Section: Article
- URL: https://journals.rcsi.science/1990-519X/article/view/212501
- DOI: https://doi.org/10.1134/S1990519X17060074
- ID: 212501
Cite item
Abstract
Proinflammatory cytokines of the interleukin-36 (IL-36) family are involved in the pathogenesis of different skin diseases in human and mice. Administration of exogenous IL-36 receptor antagonist (IL-36RA) may be an approach to therapy of different dermatitises. For its full biological activity, IL-36RA requires cleavage of N-terminal methionine residue. We created three E. coli strains producing IL-36RA coexpressed with E. coli methionine aminopeptidase under control of different promoters. To test the biological activity of IL-36RA from different strains we transfected А549 cells with plasmid carrying the IL-36 receptor gene (IL1RL2). These cells respond to IL-36g treatment with production of IL-8, which can be quantified with ELISA. IL-36RA treatment disrupts IL-36 receptor activation by IL-36g and production of IL-8. Using this system, we proved that IL-36RA from all three producer strains is fully biologically active.
About the authors
A. A. Kolobov
Institute of Highly Pure Biopreparations; Department of Biochemistry
Author for correspondence.
Email: alexey.kolobov.spb@gmail.com
Russian Federation, St. Petersburg, 197110; St. Petersburg, 199004
E. V. Kondratyeva
Institute of Highly Pure Biopreparations
Email: alexey.kolobov.spb@gmail.com
Russian Federation, St. Petersburg, 197110
T. A. Sharafutdinova
Institute of Highly Pure Biopreparations
Email: alexey.kolobov.spb@gmail.com
Russian Federation, St. Petersburg, 197110
R. S. Kalinin
Institute of Highly Pure Biopreparations; Department of Biochemistry
Email: alexey.kolobov.spb@gmail.com
Russian Federation, St. Petersburg, 197110; St. Petersburg, 199004
P. P. Nimiritsky
Department of Biochemistry
Email: alexey.kolobov.spb@gmail.com
Russian Federation, Moscow, 119192
V. E. Stefanov
Department of Biochemistry
Email: alexey.kolobov.spb@gmail.com
Russian Federation, St. Petersburg, 199004
A. V. Petrov
Institute of Highly Pure Biopreparations
Email: alexey.kolobov.spb@gmail.com
Russian Federation, St. Petersburg, 197110