Human IL-36RA production in Escherichia coli with coexpression of E. coli methionine aminopeptidase. I. Comparison of IL-36RA production in different strains
Cell and Tissue Biology(2017)
Abstract
Generalized pustular psoriasis (GPP) is a rare, sometimes lethal, form of psoriasis caused by series of mutations in the interleukin-36 receptor antagonist (IL-36RA) gene associated with its reduced expression or activity. Administration of exogenous IL-36RA can be a potent therapeutic approach to treating GPP and other forms of psoriasis. Since cleavage of the starting N-formylmethionine residue from the N-terminal end is needed for full biological activity of IL-36RA, we have developed a technique for producing IL-36RA lacking N-formylmethionine residue in E. coli . We have created a series of plasmids carrying the E. coli methionine aminopeptidase (MAP) gene under the control of different promoters for coexpression of IL-36RA and MAP and tested their effect on IL-36RA production. The highest production of IL-36RA with <3% of unprocessed molecules with uncleaved N-terminal formylmethionine residue has been shown for E. coli strain carrying the MAP gene under the control of arabinose-inducible promoter.
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Key words
Interleukin, interleukin-36 receptor antagonist (IL-36RA), methionine aminopeptidase (MAP), recombinant protein, coexpression, methionine
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