C. elegans metallothioneins: response to and defence against ROS toxicity.
C. elegans metallothioneins: response to and defence against ROS toxicity.
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DOI：
10.1039/c1mb05114h
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发表时间：
2011-08
影响因子：
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通讯作者：
Sukaina Zeitoun-Ghandour;O. I. Leszczyszyn;Claudia A. Blindauer;F. Geier;J. Bundy;S. Stürzenbaum
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文献类型：
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作者：
Sukaina Zeitoun-Ghandour;O. I. Leszczyszyn;Claudia A. Blindauer;F. Geier;J. Bundy;S. Stürzenbaum
秀丽隐杆线虫（Caenorhabditis elegans）的基因组编码两个多功能金属结合金属硫蛋白（MTs）， CeMT-1和CeMT-2。在这里，我们应用qPCR鉴定了暴露于自由基产生者（ROS）百草枯或过氧化氢后的转录上调，这一趋势与表达ppmtl::GFP的等位基因证实了。MT基因座的缺失导致体内过氧化氢水平升高，暴露于ROS导致野生型线虫的总产蛋量、生长和寿命减少，这种影响在CeMT-2和双敲除中尤为明显。此外，重组mt与过氧化氢的体外培养表明，cemt存在直接氧化，锌从两种异构体中释放出来，并伴随形成分子内二硫化物。最后，在氧化应激源存在/不存在的情况下，野生型和MT敲除的代谢谱（代谢组学）分析证实了其他实验描述的总体趋势，并确定2-氨基己二酸盐是一种潜在的新型氧化应激小分子标志物。综上所述，本研究强调秀丽隐杆线虫金属硫蛋白清除和保护活性氧和潜在的氧化应激，其中CeMT-2比CeMT-1更有效。
The genome of the nematode Caenorhabditis elegans encodes for two multifunctional metal binding metallothioneins (MTs), CeMT-1 and CeMT-2. Here we applied qPCR to identify a transcriptional up-regulation following the exposure to free radical generators (ROS) paraquat or hydrogen peroxide, a trend that was confirmed with Pmtl::GFP expressing alleles. The deletion of the MT loci resulted in an elevation of in vivo levels of hydrogen peroxide and exposure to ROS caused a reduction in total egg production, growth and life span in wild type nematodes, effects that were particularly pronounced in the CeMT-2 and double knockout. Moreover, in vitro incubation of recombinant MTs with hydrogen peroxide demonstrated the presence of direct oxidation of the CeMTs, with zinc released from both isoforms and concomitant formation of intra-molecular disulfides. Finally, metabolic profiling (metabolomic) analysis of wild type and MT knockouts in the presence/absence of oxidative stressors, confirmed the overall trend described by the other experiments, and identified 2-aminoadipate as a potential novel small-molecule marker of oxidative stress. In summary, this study highlights that C. elegans metallothioneins scavenge and protect against reactive oxygen species and potentially against oxidative stress, with CeMT-2 being more effective than CeMT-1.