Tropical Journal of Pharmaceutical Research
Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
Vol. 12, No. 3, 2013, pp. 329-334
Bioline Code: pr13051
Full paper language: English
Document type: Research Article
Document available free of charge
Tropical Journal of Pharmaceutical Research, Vol. 12, No. 3, 2013, pp. 329-334
© Copyright 2013 - Tropical Journal of Pharmaceutical Research
Expression, Purification, Characterization and In Vitro Activity of Recombinant Mouse Cu/Zn-Binding Superoxide Dismutase (mSOD1)|
Zhang, Zide; Huang, Luyuan; Luo, Zhihui; Liu, Yong; Li, Aiyun; Sun, Hua; Wu, Qiuhong; Jiang, Renwang & Wang, Feng
Purpose: To express, purify and characterize recombinant mouse Cu/Zn-binding superoxide dismutase (mSOD1), and investigate its activity in vitro.
Methods: The protein, mSOD1, was expressed after induction with isopropyl-beta-D-thiogalactopyranoside (IPTG). The target protein was purified by Ni-NTA affinity chromatography. The identity of the recombinant protein was confirmed by Western-blot and peptide mass fingerprinting (PMF) analysis. Protein activity in vitro was investigated by SOD activity assay kit and DNA damage protective assay.
Results: mSOD1 protein was expressed with a final yield of about 60 mg of pure protein per liter of culture medium. After puriﬁcation, the target protein was > 95 %. The identity of the recombinant protein was conﬁrmed. SOD activity assay showed that the highest activity of the mSOD1 was 3789.0 ± 80.5 U/mg. The present work showed that mSOD1 was effective in protecting DNA from oxidative damage.
Conclusion: High purity recombinant mSOD1 was obtained and characterized, and had high activity in vitro. The study indicates that the mSOD1 may serve as a potential therapeutic agent for those diseases caused by oxidative stress.
Cu/Zn-binding Superoxide dismutase; Expression; DNA damage; Metal ions; Purification
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