Expression, Purification and Crystal Growing Conditions of Recombinant Oryza sativa PHGPx

Chem. J. Chinese Universities

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Expression, Purification and Crystal Growing Conditions of Recombinant Oryza sativa PHGPx

WANG Feng, LIU Jin-Yuan*

Laboratory of Molecular Biology and Protein Science Laboratory of the Ministry of Education, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China

Received:2006-11-21 Revised:1900-01-01 Online:2007-09-10 Published:2007-09-10
Contact: LIU Jin-Yuan

Abstract

Abstract: Phospholipid hydroperoxide glutathione peroxidase(PHGPx) is a unique antioxidant enzyme that directly reduces lipid hydroperoxides in biomembranes. It plays potential important roles in oxidative stress response. Oryza sativa PHGPx gene was cloned into expression vector pGEX-6P-1 and transformed into E.coli strain BL21(DE3). OsPHGPx for crystals was prepared with employing Glutathione Sepharose^TM affinity, cation-exchange and gel filtration chromatography. The purity of the purified OsPHGPx was over 95%. OsPHGPx showed an obvious PHGPx activity towards lipid hydroperoxides. MALDI-TOF analysis shows that the exact molecular weight of OsPHGPx was 19275.568, which was in accordance with the theoretical molecular weight. The microcrystals of OsPHGPx were obtained under several conditions. In addition, a tertiary structure model of the OsPHGPx generated from http://swissmodel.expasv.org/ displayed the thioredoxin fold.

Key words: Oryza sativa, Phospholipid hydroperoxide glutathione peroxidase, Expression, Purification, Microcrystal growth of OsPHGPx, 3D structure modeling

CLC Number:

O650
Q518.2

TrendMD:

WANG Feng, LIU Jin-Yuan*. Expression, Purification and Crystal Growing Conditions of Recombinant Oryza sativa PHGPx[J]. Chem. J. Chinese Universities, doi: .

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Expression, Purification and Crystal Growing Conditions of Recombinant Oryza sativa PHGPx

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