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Comparative study of two thioredoxin peroxidases from disk abalone (Haliotis discus discus): cloning, recombinant protein purification, characterization of antioxidant activities and expression analysis.
Fish Shellfish Immunol. 2008 Mar; 24(3):294-307.FS

Abstract

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 1045 bp full-length cDNA with 753 and 597 bp 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 H2O2 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 H2O2 at 0.8 mM concentration by in vivo H2O2 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 H2O2 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.

Authors+Show Affiliations

Department of Marine Biotechnology, College of Ocean Science, Cheju National University, 66 Jejudaehakno, Ara-Dong, Jeju 690-756, Republic of Korea.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18226547

Citation

Pushpamali, Wickramaarachchilage Anoja, et al. "Comparative Study of Two Thioredoxin Peroxidases From Disk Abalone (Haliotis Discus Discus): Cloning, Recombinant Protein Purification, Characterization of Antioxidant Activities and Expression Analysis." Fish & Shellfish Immunology, vol. 24, no. 3, 2008, pp. 294-307.
Pushpamali WA, De Zoysa M, Kang HS, et al. Comparative study of two thioredoxin peroxidases from disk abalone (Haliotis discus discus): cloning, recombinant protein purification, characterization of antioxidant activities and expression analysis. Fish Shellfish Immunol. 2008;24(3):294-307.
Pushpamali, W. A., De Zoysa, M., Kang, H. S., Oh, C. H., Whang, I., Kim, S. J., & Lee, J. (2008). Comparative study of two thioredoxin peroxidases from disk abalone (Haliotis discus discus): cloning, recombinant protein purification, characterization of antioxidant activities and expression analysis. Fish & Shellfish Immunology, 24(3), 294-307. https://doi.org/10.1016/j.fsi.2007.11.016
Pushpamali WA, et al. Comparative Study of Two Thioredoxin Peroxidases From Disk Abalone (Haliotis Discus Discus): Cloning, Recombinant Protein Purification, Characterization of Antioxidant Activities and Expression Analysis. Fish Shellfish Immunol. 2008;24(3):294-307. PubMed PMID: 18226547.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparative study of two thioredoxin peroxidases from disk abalone (Haliotis discus discus): cloning, recombinant protein purification, characterization of antioxidant activities and expression analysis. AU - Pushpamali,Wickramaarachchilage Anoja, AU - De Zoysa,Mahanama, AU - Kang,Hyun-Sil, AU - Oh,Cheol Hong, AU - Whang,Ilson, AU - Kim,Se Jae, AU - Lee,Jehee, Y1 - 2007/12/05/ PY - 2007/10/02/received PY - 2007/11/22/revised PY - 2007/11/22/accepted PY - 2008/1/30/pubmed PY - 2008/7/25/medline PY - 2008/1/30/entrez SP - 294 EP - 307 JF - Fish & shellfish immunology JO - Fish Shellfish Immunol VL - 24 IS - 3 N2 - 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 1045 bp full-length cDNA with 753 and 597 bp 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 H2O2 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 H2O2 at 0.8 mM concentration by in vivo H2O2 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 H2O2 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. SN - 1050-4648 UR - https://www.unboundmedicine.com/medline/citation/18226547/Comparative_study_of_two_thioredoxin_peroxidases_from_disk_abalone__Haliotis_discus_discus_:_cloning_recombinant_protein_purification_characterization_of_antioxidant_activities_and_expression_analysis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1050-4648(07)00207-0 DB - PRIME DP - Unbound Medicine ER -