Abstract
Interleukin-24 (IL-24), a cytokine belonging to the IL-10 family, can selectively induce apoptosis in a broad range of tumor cells without harming normal cells. The efficient and soluble expression of bioactive recombinant IL-24 in Escherichia coli remains an obstacle because of aggregation and insufficient yield. In this study, a fusion of the small ubiquitin-related modifier (SUMO) or maltose-binding protein (MBP) has shown potential in facilitating the produce of IL-24. Thus, a new construct for MBP-SUMO-IL-24 expression would be a promising approach. Our results showed that the MBP-SUMO-IL-24 fusion protein was efficiently expressed as a soluble protein. SUMO protease-mediated cleavage at the SUMO/IL-24 junction released the recombinant IL-24 from the fusion protein. In addition, a His6 tag fused upstream of SUMO allowed for one-step purification through nickel affinity chromatography. Cleavage of the MBP-SUMO tag on the column resulted in the release of purified IL-24 and simplified the purification process. The final yield of IL-24 with approximately 90 % purity was 19 mg/L in flask fermentation. In vitro activity assays demonstrated that the purified IL-24 could induce apoptosis in MCF-7 breast cancer cells, but not normal NHLF cells, in a dose-dependent manner. In summary, we developed a novel method to express soluble and bioactive IL-24 protein in prokaryotic cells.
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Acknowledgments
We thank Chaogang Bai and Xiaojuan Wang for discussions and pointing out important references. This work was supported by the grants from the National Major Science and Technology Projects of China (Grant No. 2012ZX09304009).
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The authors declare that they have no conflict of interest.
Funding
This study was funded by the National major science and technology projects of China (Grant No. 2012ZX09304009)
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Zhang, J., Lv, X., Xu, R. et al. Soluble expression, rapid purification, and characterization of human interleukin-24 (IL-24) using a MBP-SUMO dual fusion system in Escherichia coli . Appl Microbiol Biotechnol 99, 6705–6713 (2015). https://doi.org/10.1007/s00253-015-6441-4
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DOI: https://doi.org/10.1007/s00253-015-6441-4