Expression of selenocysteine-containing glutathione S-transferase in eukaryote

Protein Expression and Purification
Huijuan LiuJingyan Wei

Abstract

Glutathione peroxidase (GPX) is a crucial antioxidant selenocysteine (Sec) containing enzyme which plays a significant role in protecting cells against oxidative damage by catalyzing the reduction of hydroperoxides with glutathione (GSH). Several methods have been used to generate GPX mimics, however, only a few of these methods involved genetic engineering and none of them have achieved specific site-directed incorporation of Sec without other modifications, which has hampered further structure-function studies. Here, we report for the first time the conversion of human glutathione transferase Zeta (hGSTZ1-1) into seleno-hGSTZ1-1 by means of genetic engineering in eukaryotes. Fluorescence microscopy images of the expression of Seleno-GST-green fluorescent protein chimaera indicated that we successfully achieved the read-through of the UGA codon to specifically incorporate Sec. Therefore, we achieved the conversion of human glutathione transferase Zeta (hGSTZ1-1) into a seleno-GST (seleno-hGSTZ1-1) by means of genetic engineering in eukaryotes. These results show that recombinant selenoproteins with incorporation of specific selenocysteine residues may be heterologously produced in eukaryotes by using a Sec insertion sequence i...Continue Reading

References

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Citations

Nov 30, 2012·Angewandte Chemie·Caroline AldagDieter Söll
Feb 14, 2015·Chemical Communications : Chem Comm·Rasa RakauskaitėSaulius Klimašauskas
May 6, 2021·Chembiochem : a European Journal of Chemical Biology·Yuchuan WangJiangyun Wang

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