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| Effects of Prokaryotic Expressed Radish Phospholipid Hydroperoxide Glutathione Peroxidase on Restoration of B16 Melanoma Cells Irradiated with Ultroviolet |
1.School of Life Sciences, Tsinghua University,Beijing 100084,China
2.Jiangsu Longliqi Bioscience Co Ltd, Changshu 215555,China |
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Abstract The radish phospholipid hydroperoxide glutathione peroxidase (RsPHGPx) and its mutant (mRsPHGPx) fusion proteins expressed in Escherichia coli BL21(DE3) with high efficiency were purified with Glutathione Sepharose 4B affinity chromatography and cleaved on-column with PreScission protease, and both the recombinant proteins without tags were obtained at a purity more than 90%. The enzymatic measurement confirmed that the activity of mRsPHGPx was far less than that of RsPHGPx. Effects of RsPHGPx and mRsPHGPx on restoration of B16 melanoma cells irradiated with ultroviolet were investigated. The results showed that the active RsPHGPx could significantly increase the cell viability and reduce the lipid peroxidation of cell membranes, while mRsPHGPx with very low activity had no effects, suggesting RsPHGPx could inhibit the ultroviolet-mediated injury in cell membranes mainly by rapidly scavenging various lipid peroxides.
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Received: 09 September 2009
Published: 26 February 2010
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