McDonagh Brian, Tyther Raymond, Sheehan David
Proteomics Research Group, Department of Biochemistry and Environmental Research Institute, University College Cork, Cork, Ireland.
Aquat Toxicol. 2005 Jul 1;73(3):315-26. doi: 10.1016/j.aquatox.2005.03.020.
Protein expression profiles (PEPs) were generated by two-dimensional electrophoresis (2-D SDS-PAGE) for gill and digestive glands of Mytilus edulis sampled from a polluted and reference site in Cork Harbour, Ireland. Similar patterns and expression levels were found for both sites in silver stained gels. However, Western blotting for carbonylated proteins demonstrated higher levels of specific carbonylation of proteins in tissues from animals in the polluted site. Animals from the reference site were acclimated in holding tanks, exposed to 1 mM H2O2 for 24 h, dissected and analysed by 2-D SDS-PAGE. Again, generally similar PEPs were found in control and exposed animals for gill and digestive gland but carbonylation was more pronounced in polluted and exposed animals. Western blotting of extracts after one-dimensional electrophoresis with antibodies to glutathione and actin revealed that gill proteins are glutathionylated more strongly than digestive gland and that this process is more pronounced in polluted animals than in controls. We conclude that carbonylation and glutathionylation can occur in gill and digestive gland in response to oxidative stress in M. edulis. Actin is a major target for both glutathionylation and carbonylation under oxidative stress conditions.
采用二维电泳（2-D SDS-PAGE）技术，对采自爱尔兰科克港污染位点和对照位点的紫贻贝鳃和消化腺进行蛋白质表达谱（PEP）分析。在银染凝胶中，两个位点的蛋白质表达模式和水平相似。然而，对羰基化蛋白质进行的蛋白质印迹分析表明，污染位点动物组织中蛋白质的特定羰基化水平更高。将对照位点的动物置于养殖水槽中驯化，暴露于1 mM过氧化氢24小时，解剖后用2-D SDS-PAGE进行分析。同样，在鳃和消化腺的对照动物和暴露动物中，通常发现相似的蛋白质表达谱，但羰基化在污染和暴露动物中更为明显。用抗谷胱甘肽和肌动蛋白抗体对一维电泳后的提取物进行蛋白质印迹分析表明，鳃蛋白的谷胱甘肽化比消化腺更强，并且这一过程在污染动物中比在对照动物中更明显。我们得出结论，紫贻贝的鳃和消化腺在氧化应激下可发生羰基化和谷胱甘肽化。在氧化应激条件下，肌动蛋白是谷胱甘肽化和羰基化的主要靶点。
