Lee Young-Mi, Lee Kyun-Woo, Park Hyun, Park Heum Gi, Raisuddin Sheikh, Ahn In-Young, Lee Jae-Seong
Department of Chemistry and the National Research Lab of Marine Molecular and Environmental Bioscience, College of Natural Sciences, Hanyang University, Seoul 133-791, South Korea.
Chemosphere. 2007 Oct;69(6):893-902. doi: 10.1016/j.chemosphere.2007.05.087. Epub 2007 Jul 19.
Glutathione S-transferases (GSTs) play a major role in detoxification of xenobiotics and antioxidant defense. Here we report full-length cDNA sequence of a novel Sigma-class of GST (GST-S) from the intertidal copepod Tigriopus japonicus. The full sequence was of 1,136 bp in length containing an open reading frame (ORF) of 651 bp that encoded 217 amino acid residues. The recombinant Tigriopus GST-S was highly expressed in transformed Escherichia coli. Kinetic properties and effects of pH, temperature and chemical inhibitors on Tigriopus GST-S were also studied. The expression of GST-S was studied using real-time RT-PCR in response to exposure to two oxidative stresses-inducing agents, viz., hydrogen peroxide (H(2)O(2)) and heavy metals (copper, manganese). It was observed that H(2)O(2) (2mM) exposure down-regulated its expression at the initial stage but there was recovery and up-regulation shortly afterwards. In case of heavy metal exposure there was concentration-dependent increase in Tigriopus GST-S gene expression up to 24h. These results suggest that Tigriopus GST-S expression is modulated by prooxidant chemicals and it may play a role against oxidative stress. A majority of other GST isoforms is known to play an important role in antioxidant defense. This study provides a preliminary insight into the possible antioxidant role for Sigma-class of GST in T. japonicus.
谷胱甘肽S-转移酶(GSTs)在异源物质的解毒和抗氧化防御中起主要作用。在此,我们报道了来自潮间带桡足类日本虎斑猛水蚤的一种新型西格玛类GST(GST-S)的全长cDNA序列。该全长序列长度为1136 bp,包含一个651 bp的开放阅读框(ORF),编码217个氨基酸残基。重组日本虎斑猛水蚤GST-S在转化的大肠杆菌中高度表达。我们还研究了日本虎斑猛水蚤GST-S的动力学特性以及pH、温度和化学抑制剂对其的影响。使用实时RT-PCR研究了GST-S在暴露于两种氧化应激诱导剂(即过氧化氢(H₂O₂)和重金属(铜、锰))时的表达情况。观察到暴露于2 mM H₂O₂在初始阶段会下调其表达,但随后不久会恢复并上调。在重金属暴露的情况下,日本虎斑猛水蚤GST-S基因表达在24小时内呈浓度依赖性增加。这些结果表明,日本虎斑猛水蚤GST-S的表达受促氧化剂化学物质调节,并且它可能在对抗氧化应激中发挥作用。已知大多数其他GST同工型在抗氧化防御中起重要作用。本研究初步揭示了西格玛类GST在日本虎斑猛水蚤中可能的抗氧化作用。