Soluble expression, rapid purification, and characterization of human interleukin-24 (IL-24) using a MBP-SUMO dual fusion system in Escherichia coli
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| 024 | 7 | 0 | |a 10.1007/s00253-015-6441-4 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6441-4 | ||
| 245 | 0 | 0 | |a Soluble expression, rapid purification, and characterization of human interleukin-24 (IL-24) using a MBP-SUMO dual fusion system in Escherichia coli |h [Elektronische Daten] |c [Jian Zhang, Xinxin Lv, Rui Xu, Xinyi Tao, Yuguo Dong, Aiyou Sun, Dongzhi Wei] |
| 520 | 3 | |a 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 19mg/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. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a Interleukin-24 (IL-24) |2 nationallicence | |
| 690 | 7 | |a Small ubiquitin-related modifier (SUMO) |2 nationallicence | |
| 690 | 7 | |a Maltose-binding protein (MBP) |2 nationallicence | |
| 690 | 7 | |a Soluble expression |2 nationallicence | |
| 690 | 7 | |a One-step purification |2 nationallicence | |
| 690 | 7 | |a Apoptosis |2 nationallicence | |
| 700 | 1 | |a Zhang |D Jian |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 700 | 1 | |a Lv |D Xinxin |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 700 | 1 | |a Xu |D Rui |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 700 | 1 | |a Tao |D Xinyi |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 700 | 1 | |a Dong |D Yuguo |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 700 | 1 | |a Sun |D Aiyou |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 700 | 1 | |a Wei |D Dongzhi |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/16(2015-08-01), 6705-6713 |x 0175-7598 |q 99:16<6705 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6441-4 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6441-4 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Zhang |D Jian |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Lv |D Xinxin |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Xu |D Rui |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Tao |D Xinyi |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Dong |D Yuguo |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sun |D Aiyou |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Wei |D Dongzhi |u State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, 200237, Shanghai, People's Republic of China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/16(2015-08-01), 6705-6713 |x 0175-7598 |q 99:16<6705 |1 2015 |2 99 |o 253 | ||