Title of article :
Expression, Purification, Characterization and In Vitro Activity of Recombinant Mouse Cu/Zn-Binding Superoxide Dismutase (mSOD1)
Author/Authors :
Zhang, Zide Jinan University - College of Pharmacy - Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, China , Huang, Luyuan Chinese Academy of Sciences - Guangzhou Institute of Biomedicine and Health, South China Institute for Stem Cell Biology and Regenerative Medicine, China , Luo, Zhihui Jinan University - College of Pharmacy, China , Liu, Yong Guangdong Institute of Eco-Environmental and Soil Sciences - Guangdong Key Laboratory of Agro-Environment Integrated Control, China , Li, Aiyun Jinan University - College of Pharmacy, China , Sun, Hua Jinan University - College of Pharmacy, China , Wu, Qiuhong Jinan University - College of Pharmacy, China , Jiang, Renwang Jinan University - College of Pharmacy, Institute of Traditional Chinese Medicine and Natural Products - Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, China , Wang, Feng Jinan University - College of Pharmacy - Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, China
Abstract :
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-Dthiogalactopyranoside (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.
Keywords :
Cu , Zn , binding Superoxide dismutase , Expression , Purification , Metal ions , DNA damage
