蛋白质组学技术筛选与鉴定在甲基对硫磷胁迫下牙鲆脑组织表达的差异蛋白质

以甲基对硫磷(MP)为有机磷农药污染源,采用蛋白质组学技术分离及鉴定在甲基对硫磷胁迫下,牙鲆(Paralichthys olivaceus)脑组织表达的差异蛋白质,从中筛选出潜在的适合于监测甲基对硫磷污染程度的蛋白指示物。实验结果表明:在甲基对硫磷胁迫下,牙鲆脑组织表达出17个差异蛋白质,经肽质量指纹(PMF)图谱技术鉴定后,发现其中部分差异蛋白质为热休克蛋白、细胞色素P450和谷胱甘肽S-转移酶,均是与受甲基对硫磷胁迫有关的蛋白质。

Differential proteins revealed with proteomics in the brain tissue of Paralichthys olivaceus under the stress of methyl parathion

The proteins from the brain tissue of the fish Paralichtys olivaceus (PO) were separated by the optimized two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Approximately 650 protein spots in each gel were detected from the brain sample when applying 200 mg protein to a 2D-PAGE gel in the pH range of 3.0-10.0. The differential proteins from the PO brain were separated and identified by proteomic technologies and they were further selected as the potential protein markers to monitor the contamination levels of methyl parathion (MP). Compared with the control group, 17 differential proteins were obtained from the PO brain under the stress of MP at 3 mg/L. Among these spots, fourteen were up-regulated and three were down-regulated. Then these differential proteins were further identified by peptide mass fingerprint (PMF) and database search. The results showed that heat shock protein 70, glutathione S-transferase and cytochrome P450 were significantly found in those differential proteins. These proteins might have strong potential for monitoring the contamination levels of MP and searching for the toxicological mechanism, as well as to evaluate the degree of risk of human fatalities under the condition of MP pollution. The results indicated that the application of the multiple biomarkers has the advantages over that of the single biomarker for monitoring the levels of MP contamination in the flowing seawater.