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RISS 인기검색어
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- 저자 : Pushpamali, W.A. (검색결과 2 건)
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Pushpamali, W.A.,De Zoysa, M.,Kang, H.S.,Oh, C.H.,Whang, I.,Kim, S.J.,Lee, J. Academic Press 2008 Fish & shellfish immunology Vol.24 No.3
Thioredoxin peroxidase (TPx), also named peroxiredoxin (Prx), is an important peroxidase, which can protect organisms against various oxidative stresses. Two TPxs were isolated from a disk abalone (Haliotis discus discus) cDNA library, named as AbTPx1 and AbTPx2, respectively. AbTPx1 and AbTPx2 consist of 1315 and 1045bp full-length cDNA with 753 and 597bp open reading frames encoding 251 and 199 amino acids, respectively. The TPx signature motif 1 (FYPLDFTFVCPTEI) and motif 2 (GEVCPA) were conserved in both AbTPx1 and AbTPx2 amino acid sequences. Purified recombinant abalone TPx fusion proteins catalyzed the reduction of H<SUB>2</SUB>O<SUB>2</SUB> and butyl hydroperoxide in peroxidase assays. Furthermore, both AbTPx fusion proteins were shown to protect super-coiled DNA from damage by metal-catalyzed oxidation (MCO) in vitro. Escherichia coli cells transformed with AbTPx1 and AbTPx2 coding sequences in pMAL-c2x showed resistance to H<SUB>2</SUB>O<SUB>2</SUB> at 0.8mM concentration by in vivo H<SUB>2</SUB>O<SUB>2</SUB> tolerance assay. AbTPx1 and AbTPx2 mRNA were constitutively expressed in gill, mantle, abductor muscle and digestive tract in a tissue specific manner. Additionally, both TPxs mRNA were up-regulated in gill and digestive tract tissues against H<SUB>2</SUB>O<SUB>2</SUB> at 3h post injection. The results indicate that AbTPx1 and AbTPx2 gene expressions are induced by oxidative stress and their respective proteins function in the detoxification of different ROS molecules to maintain efficient antioxidant defense in disk abalone.
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De Zoysa, M.,Pushpamali, W.A.,Oh, C.,Whang, I.,Kim, S.J.,Lee, J. Academic Press 2008 FISH AND SHELLFISH IMMUNOLOGY Vol.25 No.4
Selenium dependent glutathione peroxidase (Se-GPx) belongs to the family of selenoprotein, which acts mainly as an antioxidant in the cellular defence system. We have identified Se-GPx full length cDNA from disk abalone (Haliotis discus discus) designated as AbSe-GPx. It has a characteristic codon at <SUP>223</SUP>TGA<SUP>225</SUP> that corresponds to selenocysteine (Sec) amino acid as U<SUB>75</SUB>. The full length cDNA consists of 675bp, an open reading frame encoding 225 amino acids. Sequence characterization revealed that AbSe-GPx contains a characteristic GPx signature motif 2 (<SUP>97</SUP>LGFPCNQF<SUP>104</SUP>), an active site motif (<SUP>183</SUP>WNFEKF<SUP>188</SUP>) and essential residues for the enzymatic function. Additionally, the eukaryotic selenocysteine insertion sequence (SECIS) is conserved in the 3' UTR. The AbSe-GPx amino acid sequence exhibited the highest level of identity (46%) with insect (Ixodes scapularis) GPx, and shares 41% with bivalve (Unio tumidus) Se-GPx. The RT-PCR analysis revealed that AbSe-GPx mRNA was expressed constitutively in gill, mantle, gonad, abductor muscle, digestive tract, and hemocytes in a tissue specific manner. AbSe-GPx mRNA expression was significantly up-regulated in gill and digestive tract tissues after H<SUB>2</SUB>O<SUB>2</SUB> injection and Vibrio alginolyticus infection. However, AbSe-GPx expression was not up-regulated after Aroclor 1254 injection. These results indicate that AbSe-GPx mRNA is expressed at a basal level in abalone tissues, which can be up-regulated transcriptionally by H<SUB>2</SUB>O<SUB>2</SUB> oxidative stress and Vibrio alginolyticus infection. Therefore, AbSe-GPx may be involved in a protective role against H<SUB>2</SUB>O<SUB>2</SUB> oxidative stress and immune defence against bacterial infection.
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