Human IL-36RA production in Escherichia coli with coexpression of E. coli methionine aminopeptidase. II. Comparison of IL-36RA biological activity from different strains

A. A. Kolobov, E. V. Kondratyeva, T. A. Sharafutdinova,R. S. Kalinin,P. P. Nimiritsky, V. E. Stefanov,A. V. Petrov

Cell and Tissue Biology(2017)

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摘要
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.
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关键词
interleukin, Interleukin-36 receptor antagonist (IL-36RA), methionine aminopeptidase (MAP), recombinant protein, coexpression, methionine, А549 cells
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