Skip to main content Accessibility help
×
Home

Expression and characterization of a phospholipid hydroperoxide glutathione peroxidase gene in Schistosoma japonicum

  • YING ZHANG (a1), YUAN HE (a2), LI HE (a2), HONGYING ZONG (a2) and GUOBIN CAI (a2)...

Summary

Phospholipid hydroperoxide glutathione peroxidase (PHGPx, GPx4) is a major antioxidant enzyme, which plays unique roles in the protection of cells against oxidative stress by catalysing reduction of lipid hydroperoxides. We isolated and characterized a full-length cDNA sequence encoding GPx gene from a blood fluke, Schistosoma japonicum (designated SjGPx), which contained an in-frame TGA codon for selenocysteine (Sec) and a concurrent Sec insertion sequence in its 3′-untranslated region. Protein encoded by SjGPx demonstrated a primary structure characteristic to the PHGPx family, including preservation of catalytic domains and absence of the subunit interaction domains. Semi-quantitative reverse transcription PCR and Western blotting showed that the SjGPx was mainly expressed in the female adults and eggs. RNA interference approach was employed to investigate the effects of knockdown of SjGPx. SjGPx expression level was significantly reduced on the 5th day post-RNAi. Significantly reduction in GPx enzyme activities, as well as obvious changes in morphology of intrauterine eggs followed the reduction in SjGPx transcript level. We observed a 63·04% reduction in GPx activity and the eggs severely deformed. Our results revealed that SjGPx protein might be involved in the provision of enzyme activity during egg production.

Copyright

Corresponding author

* Corresponding author. Department of Medical Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China. E-mail: gbcai2011@whu.edu.cn

References

Hide All
Arthur, J. R. (2000). The glutathione peroxidases. Cellular and Molecular Life Sciences 57, 18251835.
Bae, Y. A., Kim, S. H., Ahn, C. S., Kim, J. G. and Kong, Y. (2015). Molecular and biochemical characterization of Paragonimus westermani tyrosinase. Parasitology 142, 807815.
Brigelius-Flohé, R., Aumann, K. D., Blöcker, H., Gross, G., Kiess, M., Klöppel, K. D., Maiorino, M., Roveri, A., Schuckelt, R., Usani, F., Wingender, E. and Flohé, L. (1994). Phospholipid-hydroperoxide glutathione peroxidase. Genomic DNA, cDNA, and deduced amino acid sequence. Journal of Biological Chemistry 269, 73427348.
Cai, G., Bae, Y., Zhang, Y., He, Y., Jiang, M. and He, L. (2009). Expression and characterization of two tyrosinases from the trematode Schistosoma japonicum . Parasitology Research 104, 601609.
Cai, G. B., Bae, Y. A., Kim, S. H., Sohn, W. M., Lee, Y. S., Jiang, M. S., Kim, T. S. and Kong, Y. (2008). Vitellocyte-specific expression of hydroperoxide glutathione peroxidases in Clonorchis sinensis . International Journal for Parasitology 38, 16131623.
Callahan, H. L., Crouch, R. K. and James, E. R. (1988). Helminth anti-oxidant enzymes: a protective mechanism against host oxidants? Parasitology Today 4, 218225.
Cheng, G., Fu, Z., Lin, J., Shi, Y., Zhou, Y., Jin, Y. and Cai, Y. (2009). In vitro and in vivo evaluation of small interference RNA-mediated gynaecophoral canal protein silencing in Schistosoma japonicum . The Journal of Gene Medicine 11, 412421.
Cordingley, J. S. (1987). Trematode eggshells: novel protein biopolymers. Parasitology Today 22, 341344.
Dröge, W. (2002). Free radicals in the physiological control of cell function. Physiological Reviews 82, 4795.
Epp, O., Ladenstein, R. and Wendel, A. (1983). The refined structure of the selenoenzyme glutathione peroxidase at 0·2-nm resolution. European Journal of Biochemistry 133, 5169.
Hancok, J. T., Desikan, R. and Neill, S. J. (2001). Role of reactive oxygen species in cell signalling pathways. Biochemistry Society Transactions 29, 345350.
Henkle-Dührsen, K. and Kampkötter, A. (2001). Antioxidant enzyme families in parasitic nematodes. Molecular and Biochemical Parasitology 114, 129142.
He, Y., Cai, G., Ni, Y., Li, Y., Zong, H. and He, L. (2012). siRNA-mediated knockdown of two tyrosinase genes from Schistosoma japonicum cultured in vitro . Experimental Parasitology 132, 394402.
Imai, H. and Nakagawa, Y. (2003). Biological significance of phospholipids hydroperoxide glutathione peroxidase (PHGPx, GPx4) in mammalian cells. Free Radical Biology and Medicine 34, 145169.
Kim, S. H., Cai, G. B., Bae, Y. A., Lee, E. G., Lee, Y. S. and Kong, Y. (2009). Two novel phospholipid hydroperoxide glutathione peroxidase genes of Paragonimus westermani induced by oxidative stress. Parasitology 136, 553565.
Krautz-Peterson, G., Radwanska, M., Ndegwa, D., Shoemaker, C. B. and Skelly, P. J. (2007). Optimizing gene suppression in schistosomes using RNA interference. Molecular and Biochemical Parasitology 2, 194202.
Kryukov, G. V., Castellano, S., Novoselov, S. V., Lobanov, A. V., Zehtab, O., Guigó, R. and Gladyshev, V. N. (2003). Characterization of mammalian selenoproteomes. Science 300, 14391443.
Kumagai, T., Osada, Y., Ohta, N. and Kanazawa, T. (2009). Peroxiredoxin-1 from Schistosoma japonicum functions as a scavenger against hydrogen peroxide but not nitric oxide. Molecular and Biochemical Parasitology 164, 2631.
McManus, D. P., Hu, W., Brindley, P. J., Feng, Z. and Han, Z. G. (2004). Schistosome transcriptome analysis at the cutting edge. Trends in Parasitology 20, 301304.
Mei, H. and LoVerde, P. T. (1995). Schistosoma mansoni: cloning the gene encoding glutathione peroxidase. Experimental Parasitology 80, 319322.
Mei, H., Thakur, A., Schwarts, J. and LoVerde, P. T. (1996). Expression and characterization of glutathione peroxidase activity in the human blood fluke Schistosoma mansoni . Infection Immunity 64, 42994306.
Mkoji, G. M., Smith, J. M. and Prichard, R. K. (1988). Antioxidant systems in Schistosoma mansoni: correlation between susceptibility to oxidant killing and the levels of scavengers of hydrogen peroxide and oxygen free radicals. International Journal for Parasitology 18, 661666.
Morales, M. E., Rinaldi, G., Gobert, G. N., Kines, K. J., Tort, J. F. and Brindley, P. J. (2008). RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade. Molecular and Biochemical Parasitology 157, 160168.
Mourão, M. M., Dinguirard, N., Franco, G. R. and Yoshino, T. P. (2009). Phenotypic screen of early developing larvae of the blood Fluke, Schistosoma mansoni, using RNA interference. PLoS Neglected Tropical Diseases 3, e502.
Roche, C., Liu, J. L., LePresle, T., Capron, A. and Pierce, R. J. (1996). Tissue localization and stage-specific expression of the phospholipid hydroperoxide glutathione peroxidase of Schistosoma mansoni . Molecular and Biochemical Parasitology 75, 187195.
Selkirk, M. E., Smith, V. P., Thomas, G. R. and Gounaris, K. (1998). Resistance of filarial nematode parasites to oxidative stress. International Journal for Parasitology 28, 13151332.
Sies, H. (1993). Strategies of antioxidant defense. European Journal of Biochemistry 215, 213219.
Stadtman, T. C. (1996). Selenocysteine. Annual Review of Biochemistry 65, 83100.
Tabara, H., Grishok, A. and Mello, C. C. (1998). RNAi in C. elegans: soaking in the genome sequence. Science 282, 430431.
Tavernarakis, N., Wang, S. L., Dorovkov, M., Ryazanov, A. and Driscoll, M. (2000). Heritable and inducible genetic interference by double-stranded RNA encoded by transgenes. Nature Genetics 2, 180183.
Tchoubrieva, E. B., Ong, P. C., Pike, R. N., Brindley, P. J. and Kalinna, B. H. (2010). Vector-based RNA interference of cathepsin B1 in Schistosoma mansoni . Cellular and Molecular Life Sciences 67, 37393748.
Ursini, F. and Bindoli, A. (1987). The role of selenium peroxidases in the protection against oxidative damage of membranes. Chemistry and Physics of Lipids 44, 255276.
Williams, D. L., Pierce, R. J., Cookson, E. and Capron, A. (1992). Molecular cloning and sequencing of glutathione peroxidase from Schistosoma mansoni . Molecular and Biochemical Parasitology 52, 127130.
Wyler, D. J. (1992). Why does liver fibrosis occur in schistosomiasis? Parasitology Today 8, 277279.
Zamore, P. D., Tuschl, T., Sharp, P. A. and Bartel, D. P. (2000). RNAi: double-stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell 101, 2533.
Zelck, U. E. and von Janowsky, B. (2004). Antioxidant enzymes in intramolluscan Schistosoma mansoni and ROS-induced changes in expression. Parasitology 128, 493501.

Keywords

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed