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微粒体谷胱甘肽转移酶1减轻了活性氧诱导的刺参脂质过氧化反应。

Microsomal glutathione transferase 1 attenuated ROS-induced lipid peroxidation in Apostichopus japonicus.

作者信息

Zhang Zhen, Lv Zhimeng, Shao Yina, Qiu Qiongfen, Zhang Weiwei, Duan Xuemei, Li Ye, Li Chenghua

机构信息

School of Marine Sciences, Ningbo University, Ningbo 315211, PR China.

School of Marine Sciences, Ningbo University, Ningbo 315211, PR China.

出版信息

Dev Comp Immunol. 2017 Aug;73:79-87. doi: 10.1016/j.dci.2017.03.011. Epub 2017 Mar 14.

Abstract

Microsomal glutathione transferase (mGST) is a membrane bound glutathione transferase in multifunctional detoxification isoenzymes family and also plays crucial roles in innate immunity. In the present study, a novel microsomal GST homology was identified from Apostichopus japonicus (designated as AjmGST1) by RACE approaches. The full-length cDNA of AjmGST1 was of 1296 bp encoded a protein of 169 amino acids residues. Multiple sequence alignment and phylogenetic analysis together supported that AjmGST1 belonged to a new member in invertebrates mGST family. Spatial expression analysis revealed that AjmGST1was ubiquitously expressed in all examined tissues with the larger magnitude in tentacle. Time-course expression of AjmGST1 mRNA in coelomocytes was up-regulated after Vibrio splendidus challenge from 6 h until 72 h with the peak expression in 24 h, compared with that in the control group. Similarly, the induced expression of AjmGST1 expression was also detected in lipopolysaccharide (LPS) exposed primary coelomocytes. The purified recombinant protein of AjmGST1 showed high activity with GST substrate at pH of 7.0 and temperature of 35 °C. Meantime, the recombinant AjmGST1 depressed HO-induced MDA production both in vivo and in vitro. All of these results indicated that AjmGST1 was an important regulator in elimination of lipid peroxidation under immune response.

摘要

微粒体谷胱甘肽转移酶(mGST)是多功能解毒同工酶家族中的一种膜结合谷胱甘肽转移酶,在先天免疫中也发挥着关键作用。在本研究中,通过RACE方法从刺参中鉴定出一种新的微粒体GST同源物(命名为AjmGST1)。AjmGST1的全长cDNA为1296 bp,编码一个由169个氨基酸残基组成的蛋白质。多重序列比对和系统发育分析共同支持AjmGST1属于无脊椎动物mGST家族的一个新成员。空间表达分析显示,AjmGST1在所有检测组织中均有广泛表达,在触手组织中表达量较高。与对照组相比,灿烂弧菌攻击后,体腔细胞中AjmGST1 mRNA的时序表达在6小时至72小时内上调,在24小时达到峰值表达。同样,在暴露于脂多糖(LPS) 的原代体腔细胞中也检测到AjmGST1的诱导表达。纯化的重组AjmGST1蛋白在pH为7.0、温度为35℃时对GST底物表现出高活性。同时,重组AjmGST1在体内和体外均能抑制HO诱导的MDA产生。所有这些结果表明,AjmGST1是免疫反应中消除脂质过氧化的重要调节因子。

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