Abstract
Excessive production of transforming growth factor-β1 (TGF-β1) and its binding to transforming growth factor-β receptor type II (TGF-βRII) promotes fibrosis by activation of the TGF-β1-mediated signaling pathway. Thus, the truncated extracellular domain of TGF-βRII (tTβRII) is a promising anti-fibrotic candidate, as it lacks the signal transduction domain. In this work, the native N-terminal tTβRII was prepared as a His-SUMO fusion protein (termed His-SUMO-tTβRII) in Escherichia coli strain BL21 (DE3). His-SUMO-tTβRII was expressed as a soluble protein under optimal conditions (6 h of induction with 0.5 mM IPTG at 37 °C). His-SUMO-tTβRII was purified by Ni–NTA resin chromatography, and then cleaved with SUMO protease to release native tTβRII, which was re-purified using a Ni–NTA column. Approximately 12 mg of native tTβRII was obtained from a one liter fermentation culture with no less than 95% purity. In vivo studies demonstrated that tTβRII prevented CCl4-induced liver fibrosis, as evidenced by the inhibition of fibrosis-related Col I and α-SMA protein expression in C57BL/6 mice. In addition, tTβRII downregulated phosphorylation of SMAD2/3, which partly repressed TGF-β1-mediated signaling. These data indicate that the His-SUMO expression system is an efficient approach for preparing native tTβRII that possesses anti-liver fibrotic activity, allowing for the large-scale production of tTβRII, which potentially could serve as an anti-fibrotic candidate for treatment of TGF-β1-related diseases.
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Acknowledgements
This study was supported by Grants from National Natural Sciences Foundation of China (Nos. 81500471, 81700544), University Nursing Program for Young Scholars with Creative Talents in Heilongjiang Province (No. UNPYSCT-2016112), Basic Scientific Research Project of University belongs to Heilongjiang in 2017 under Grant (Nos. 2017-KYYWF-0649, 2017-KYYWF-0640) and Science and Technology Project belongs to Mudanjiang (No. Z2017s0031).
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Wang, X., Li, Y., Li, X. et al. Expression, purification, and evaluation of in vivo anti-fibrotic activity for soluble truncated TGF-β receptor II as a cleavable His-SUMO fusion protein. World J Microbiol Biotechnol 34, 181 (2018). https://doi.org/10.1007/s11274-018-2565-x
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DOI: https://doi.org/10.1007/s11274-018-2565-x