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| Expression, Purification, Activity Detection and Crystal Growth Studies on Sf2523 Protein from Shigella flexneri 2a Strain 301 |
| FENG Duo1, ZHANG Kuo2, LI Qian-qian1, HAN Cui-xiao1, GAO Wei 1 |
1. National Engineering Laboratory for Tree Breeding /College of Science, Beijing Forestry University, Beijing 100083, China;
2. Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China |
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Abstract Sf2523 protein is a member from the Thioredoxin Peroxidase (Prx) family. Peroxiredoxins are ubiquitous proteins that catalyze the reduction of hydroperoxides, thus conferring resistance to oxidative stress. Sf2523 protein can protect the large biological molecules from reactive oxygen specie, playing an important role during aerobic metabolism. After the construction of prokaryotic expression system, the Sf2523 protein was expressed in soluble state, and purified by Ni Affinity Chromatography and Size Exclusive Chromatography. The peroxidase activity experiment proved that it still has a natural enzyme activity and the core structure has not changed. Enzyme protein in vivo and in vitro has different state of aggregation. The purified monomeric protein of crystallization experiments, screening out the needle-like crystals. Through further resection of label protein to optimize the processing, finally, a homogeneous three-dimensional crystal was got.
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Received: 19 April 2012
Published: 25 August 2012
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