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Molecular and Catalytic Properties of Glutathione Peroxidase Family Proteins from Pinus tabulaeformis

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Abstract

Glutathione peroxidase (GPX) is one of the key enzymes that protect cells against oxidative damage caused by reactive oxygen species. Previous studies of plant GPXs focused mainly on angiosperms. In contrast, little information is available on the molecular characteristics of this gene family in gymnosperms. In this study, four GPX genes (PtaGPX1, 2, 3, and 4) were cloned from the gymnosperm Pinus tabulaeformis, which showed high protein sequence identity and similar expression patterns in various tissues. The four Pinus GPX proteins were expressed in Escherichia coli, and the purified proteins used thioredoxin, but not glutathione, as an electron donor. The four Pinus GPXs showed different enzymatic activities and kinetic characteristics, suggesting functional divergence. Two conserved Cys residues (corresponding to Cys44 and Cys92 of PtaGPX3) were identified in all plant GPXs, and their functions were assessed using site-directed mutagenesis. Cys44 and Cys92 of PtaGPX3 could form an intramolecular disulfide bond under oxidizing conditions. These two residues were critical components of active sites and contributed to catalytic activity. This study provides novel insights into the functional divergence and catalytic properties of the GPX family in gymnosperms.

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Acknowledgments

This study was supported by grants from the Natural Science Foundation of China (NSFC 31270641).

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Authors and Affiliations

  1. College of Life Sciences and Biotechnology, Beijing Forestry University, 100083, Beijing, China

    Li Zhao, Xue-Min Han, Wei Wang & Hai-Ling Yang

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  1. Li Zhao
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  2. Xue-Min Han
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  3. Wei Wang
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  4. Hai-Ling Yang
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Corresponding author

Correspondence to Hai-Ling Yang.

Additional information

Li Zhao and Xue-Min Han contributed equally to this work.

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Zhao, L., Han, XM., Wang, W. et al. Molecular and Catalytic Properties of Glutathione Peroxidase Family Proteins from Pinus tabulaeformis . Plant Mol Biol Rep 32, 771–778 (2014). https://doi.org/10.1007/s11105-013-0692-y

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