Biochemical characterization of human peroxiredoxin 2, an antioxidative protein.
Human peroxiredoxin 2 (Prx2), which is abundant in erythrocytes, has been shown to play a key role in protecting erythrocytes against oxidative stress by scavenging reactive oxygen species as well as participating in cell signal transduction. Here, human Prx2 gene was successfully cloned into Escherichia coli BL21 (DE3) for Prx2 expression. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis suggested that the recombinant protein was expressed mainly in a soluble form. The recombinant protein was purified by one-step Ni-nitrilotriacetic acid chelating affinity chromatography to a purity of up to 91.5%. The peroxidase activity of Prx2 to scavenge H(2)O(2) was determined by a ferrithiocyanate assay. The ability of Prx2 to protect plasmid DNA was tested by using a mixed-function oxidation system, and results showed that Prx2 could prevent DNA from undergoing oxidative stress. Ultraviolet (UV)-induced cell apoptosis assay demonstrated that Prx2 is also able to protect NIH/3T3 cells from UV-induced damage, suggesting its possible applications in cosmetics and other areas.
School of Bioscience and Bioengineering,  South China University of Technology, Guangzhou 510006,  China.
Sheng
S
Yan
Shaopei
S
Chen
Zhendong
Z
Li
Haiying
H
Wang
Tuxiong
T
Huang
Xiaoning
X
Wang
Jufang
J
Wang
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D017209:Apoptosis / Q000528:radiation effects; D015153:Blotting, Western; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D051379:Mice; D041681:NIH 3T3 Cells; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D009195:Peroxidase / Q000235:genetics / Q000378:metabolism*; D054464:Peroxiredoxins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014466:Ultraviolet Rays
D000975:Antioxidants; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D004247:DNA; D006861:Hydrogen Peroxide; D054464:Peroxiredoxins; D009195:Peroxidase