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Truncated chicken interleukin-1beta with increased biologic activity.

Zsuzsa Gyorfy·Annette Ohnemus·Bernd Kaspers·Erno Duda·Peter Staeheli
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Abstract

Chicken interleukin-1beta (ChIL-1beta) is synthesized as a precursor molecule that unlike its mammalian counterpart, lacks a typical caspase-1 cleavage site. Therefore, it was unclear if proteolytic cleavage of ChIL-1beta can occur and if cleavage might modulate the biologic activity of this cytokine. Using an avian indicator cell line that carries an NF-kappaB-regulated luciferase reporter gene, we established a sensitive and highly specific bioassay for ChIL-1beta. Experiments with a rabbit antiserum indicated that the NF-kappaB-stimulating activity in supernatants of lipopolysaccharide (LPS)-treated chicken HD-11 macrophages is largely due to IL-1beta and that proteolytic processing of natural and recombinant ChIL-1beta is not very efficient. Functional analyses further revealed that cDNAs for either full-length or N-terminally truncated chicken ChIL-1beta yielded active cytokine. A truncated molecule that closely resembled putative mature ChIL-1beta exhibited more than 100-fold enhanced biologic activity after expression in mammalian cells, indicating that precursor cleavage is indeed of critical importance for maximal activity.

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AnimalsBinding SitesBiological AssayCell LineChickensDNA, ComplementaryGenes, ReporterHumansInterleukin-1LuciferasesNF-kappa BProtein Processing, Post-TranslationalQuailRecombinant ProteinsJournal ArticleResearch Support, Non-U.S. Gov't

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Zsuzsa Gyorfy, Annette Ohnemus, Bernd Kaspers, Erno Duda, Peter Staeheli. 2003. Truncated chicken interleukin-1beta with increased biologic activity.. https://doi.org/10.1089/107999003321829935

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