Analysis of the expression and antioxidative property of a peroxiredoxin 6 from Scophthalmus maximus.
Peroxiredoxins (Prxs) are a group of antioxidant proteins that protect cells from oxidative damage caused by various peroxides. To date, six different isoforms of peroxiredoxin (Prx1 to Prx6) have been identified, of which, Prx6 belongs to the 1-Cys Prx subfamily. Although Prx6 of several fish species have been reported at sequence level, there are very few documented studies on the potential function of fish Prx6. In this report, we describe the identification and analysis of a Prx6 homologue, SmPrx6, from turbot Scophthalmus maximus. The full length cDNA of SmPrx6 contains a 5'- untranslated region (UTR) of 60 bp, an open reading frame of 666 bp, and a 3'-UTR of 244 bp. The deduced amino acid sequence of SmPrx6 shares 81-87% overall identities with known fish Prx6. In silico analysis identified in SmPrx6 a conserved Prx6 catalytic motif, PVCTTE, and the catalytic triads putatively involved in peroxidase and phospholipase A2 activities. Expression of SmPrx6 was detected in most fish organs, with the highest expression levels found in blood and heart and the lowest level in spleen. Experimental challenges with bacterial pathogens and poly(I:C) upregulated SmPrx6 expression in liver and spleen in a manner that is dependent on the challenging agent and the tissue type. Treatment of cultured primary hepatocytes with H(2)O(2) enhanced SmPrx6 expression in a dose-dependent manner. Recombinant SmPrx6 expressed in and purified from Escherichia coli exhibited thiol-dependent antioxidant activity and could protect cultured hepatocytes from H(2)O(2)-induced oxidative damage. Taken together, these results indicate that SmPrx6 is a Prx6 homologue with antioxidative property and is likely to be involved in both cellular maintenance and protective response during host immune defense against bacterial infection.
Zheng
Wen-jiang
WJ
Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, PR China.
Hu
Yong-hua
YH
Zhang
Min
M
Sun
Li
L
D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000302:isolation & purification / Q000494:pharmacology; D001483:Base Sequence; D002478:Cells, Cultured; D005393:Fish Diseases / Q000276:immunology; D005412:Flatfishes / Q000235:genetics / Q000276:immunology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016905:Gram-Negative Bacterial Infections / Q000276:immunology / Q000662:veterinary; D022781:Hepatocytes / Q000187:drug effects; D007369:Interferon Inducers / Q000494:pharmacology; D053526:Listonella / Q000276:immunology; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000187:drug effects; D054465:Peroxiredoxin VI / Q000235:genetics* / Q000276:immunology*; D011070:Poly I-C / Q000494:pharmacology; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D013290:Streptococcal Infections / Q000276:immunology / Q000662:veterinary; D013291:Streptococcus / Q000276:immunology
D000975:Antioxidants; D007369:Interferon Inducers; D011994:Recombinant Proteins; D054465:Peroxiredoxin VI; D011070:Poly I-C