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多毛纲动物琥珀新糠虾中θ类谷胱甘肽S-转移酶(GST-T)的分子克隆、表达、生化特性及生物标志物潜力

Molecular cloning, expression, biochemical characteristics, and biomarker potential of theta class glutathione S-transferase (GST-T) from the polychaete Neanthes succinea.

作者信息

Rhee Jae-Sung, Lee Young-Mi, Hwang Dae-Sik, Won Eun-Ji, Raisuddin Sheikh, Shin Kyung-Hoon, Lee Jae-Seong

机构信息

Department of Molecular and Environmental Bioscience, Graduate School, Hanyang University, Seoul, South Korea.

出版信息

Aquat Toxicol. 2007 Jun 15;83(2):104-15. doi: 10.1016/j.aquatox.2007.03.015. Epub 2007 Mar 27.

Abstract

We cloned and sequenced the full-length cDNA of a theta class glutathione S-transferase (GST-T) from the polychaete Neanthes succinea. The open reading frame of N. succinea GST-T cDNA was 678bp and encoded 226 amino acid residues. We generated recombinant N. succinea GST-T by expression in transformed Escherichia coli and studied the kinetic properties as well as the effects of inhibitors, pH, and temperature on N. succinea GST-T. GST-T expression was studied using real-time RT-PCR in response to exposure to the model oxidative stress-inducing agent, CuCl(2). Copper induced a concentration-dependant increase in the expression of GST-T. Moreover, polychaetes collected from a heavily contaminated lake near an industrial complex showed significantly higher levels of GST-T expression. Interestingly, the site-collected polychaetes with the highest GST-T mRNA expression levels also showed the highest metallothioneins levels. These results suggest that GST-T in polychaetes may have an antioxidant role and that N. succinea GST-T expression may be a useful biomarker for exposure to environmental contaminants such as copper. Our findings provide a better understanding of the biochemical characteristics of N. succinea GST-T, and elucidate the potential role of GST-T in heavy metal-induced oxidative stress and as a biomarker for environmental contamination.

摘要

我们克隆并测序了多毛纲动物琥珀新米虾(Neanthes succinea)中一种θ类谷胱甘肽S-转移酶(GST-T)的全长cDNA。琥珀新米虾GST-T cDNA的开放阅读框为678bp,编码226个氨基酸残基。我们通过在转化的大肠杆菌中表达来生成重组琥珀新米虾GST-T,并研究了其动力学特性以及抑制剂、pH值和温度对琥珀新米虾GST-T的影响。使用实时RT-PCR研究了GST-T在暴露于模型氧化应激诱导剂氯化铜(CuCl₂)时的表达情况。铜诱导了GST-T表达的浓度依赖性增加。此外,从工业园区附近一个重度污染湖泊采集的多毛纲动物显示出显著更高水平的GST-T表达。有趣的是,采集地点中GST-T mRNA表达水平最高的多毛纲动物也显示出最高的金属硫蛋白水平。这些结果表明,多毛纲动物中的GST-T可能具有抗氧化作用,并且琥珀新米虾GST-T表达可能是暴露于铜等环境污染物的一种有用生物标志物。我们的研究结果更好地理解了琥珀新米虾GST-T的生化特性,并阐明了GST-T在重金属诱导的氧化应激中的潜在作用以及作为环境污染生物标志物的作用。

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