Molecular cloning and functional characterization of theta class glutathione S-transferase from Apostichopus japonicus

作者：Shao, Yina; Lv, Zhimeng; Li, Chenghua^*; Zhang, Weiwei; Duan, Xuemei; Qiu, Qiongfen; Jin, Chunhua; Zhao, Xuelin

来源：Fish & Shellfish Immunology, 2017, 63: 31-39.

DOI：10.1016/j.fsi.2017.02.004

• Apostichopus japonicus
• Glutathione S-transferase
• Gene expression
• Immune response

Glutathione S-transferases (GSTs) are the superfamily of multifunctional detoxification isoenzymes and play crucial roles in innate immunity. In the present study, a theta class GST homology was identified from A. japonicus (designated as AjGST-theta) by RACE approaches. The full-length cDNA of AjGST-theta was of 1013 bp encoded a cytosolic protein of 231 amino acids residues. Structural analysis revealed that AjGST-theta processed the characteristic N-terminal GSH-binding site (G-site) and the C-terminal hydrophobic substrate binding site (H-site). Multiple sequence alignment and phylogenetic analysis together supported that AjGST-theta belonged to a new member of theta class GST protein subfamily. Spatial expression analysis revealed that AjGST-theta was ubiquitously expressed in all examined tissues with the larger magnitude in intestine. The Vibrio splendidus challenge in vivo and LPS stimulation in vitro could both significantly up-regulate the mRNA expression of AjGST-theta when compared with control group. The recombinant protein was expressed in Escherichia coli and the purified AjGST-theta showed high activity with GST substrate. Meantime, disc diffusion assay showed that recombinant AjGST-theta protein could markedly improve bacterial growth under Cumene hydroperoxide exposure. More importantly, the recombinant AjGST-theta could effectively prevent primary coelomocytes apoptosis after LPS exposure. Our present findings suggested that AjGST-theta might play significantly roles in the modulation of immune response and protect cells from pathogens infection in A. japonicus.

• 出版日期2017-4
• 单位宁波大学