Biochemical characterization of human peroxiredoxin 2, an antioxidative protein

Biochemical characterization of human peroxiredoxin 2, an antioxidative protein

Sheng Yan;Shaopei Chen;Zhendong Li;Haiying Wang;Tuxiong Huang;Xiaoning Wang;Jufang Wang

【期刊名称】《生物化学与生物物理学报：英文版》 【年(卷),期】2012(044)009

【摘要】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 H2O2was 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