First report of a thioredoxin homologue in jellyfish: molecular cloning, expression and antioxidant activity of CcTrx1 from Cyanea capillata.
Thioredoxins (Trx proteins) are a family of small, highly-conserved and ubiquitous proteins that play significant roles in the resistance of oxidative damage. In this study, a homologue of Trx was identified from the cDNA library of tentacle of the jellyfish Cyanea capillata and named CcTrx1. The full-length cDNA of CcTrx1 was 479 bp with a 312 bp open reading frame encoding 104 amino acids. Bioinformatics analysis revealed that the putative CcTrx1 protein harbored the evolutionarily-conserved Trx active site 31CGPC34 and shared a high similarity with Trx1 proteins from other organisms analyzed, indicating that CcTrx1 is a new member of Trx1 sub-family. CcTrx1 mRNA was found to be constitutively expressed in tentacle, umbrella, oral arm and gonad, indicating a general role of CcTrx1 protein in various physiological processes. The recombinant CcTrx1 (rCcTrx1) protein was expressed in Escherichia coli BL21 (DE3), and then purified by affinity chromatography. The rCcTrx1 protein was demonstrated to possess the expected redox activity in enzymatic analysis and protection against oxidative damage of supercoiled DNA. These results indicate that CcTrx1 may function as an important antioxidant in C. capillata. To our knowledge, this is the first Trx protein characterized from jellyfish species.
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Zengliang
Z
Ruan
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Guoyan
G
Liu
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Yufeng
Y
Guo
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Yonghong
Y
Zhou
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Qianqian
Q
Wang
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Yinlong
Y
Chang
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Beilei
B
Wang
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Jiemin
J
Zheng
Department of Marine Biotechnology, Faculty of Naval Medicine, Second Military Medical University, Shanghai, China.
Liming
L
Zhang
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010084:Oxidation-Reduction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D037401:Scyphozoa / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
D018076:DNA, Complementary; D011994:Recombinant Proteins; D013879:Thioredoxins