Molecular cloning and functional characterization of theta class glutathione S-transferase from Apostichopus japonicus. Issue 63 (April 2017)

• Record Type: Journal Article
• Authors: Shao, Yina; Lv, Zhimeng; Li, Chenghua; Zhang, Weiwei; Duan, Xuemei; Qiu, Qiongfen; Jin, Chunhua; Zhao, Xuelin
• Is Part Of: Fish & shellfish immunology. Issue 63(2017)
• Page Start: 31
• Page End: 39
• Publication Date: 2017-04
• Subjects: Apostichopus japonicus -- Glutathione S-transferase -- Gene expression -- Immune response; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2017.02.004
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: 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-θ ) by RACE approaches. The full-length cDNA of AjGST-θ was of 1013 bp encoded a cytosolic protein of 231 amino acids residues. Structural analysis revealed that AjGST-θ 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-θ belonged to a new member of theta class GST protein subfamily. Spatial expression analysis revealed that AjGST-θ 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-θ when compared with control group. The recombinant protein was expressed in Escherichia coli and the purified AjGST-θ showed high activity with GST substrate. Meantime, disc diffusion assay showed that recombinant AjGST-θ protein could markedly improve bacterial growth under Cumene hydroperoxide exposure. More importantly, the recombinant AjGST-θ could effectively prevent primary coelomocytes apoptosis after LPS exposure. Our present findings suggested that AjGST-θ might play significantly roles in the modulation of immune response and protect cells from pathogens infection in A. japonicus . Highlights: The full-length cDNA of AjGST-θ was identified in Apostichopus japonicus. AjGST-θ 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 AjGST-θ mRNA expression. The recombinant AjGST-θ could markedly improve bacterial growth under Cumene hydroperoxide exposure. The recombinant AjGST-θ could effectively prevent primary coelomocytes apoptosis in LPS exposed coelomocytes.