Isolation, Identification and Bioinformatics Analysis of Phosphatidy Lethanolamine-binding Protein 1 (PEBP1) Associated with Liver Toxicity Induced by Copper Nanoparticles in Rats (Rattus norvegicus)
摘要为筛选纳米铜对大鼠(Rattus norvegicus)肝脏毒性的差异蛋白，探讨其毒性作用机制，本研究应用双向电泳(two dimensional gel electro-phoresis, 2-DE)和质谱等蛋白质组学方法，分离和鉴定肝毒性相关差异蛋白，并利用荧光定量PCR验证和生物信息学分析。结果共筛选到显著差异表达的蛋白斑点共43个，其中下调的一个差异蛋白点1006被鉴定为磷脂酰乙醇胺结合蛋白1(phosphatidy lethanolamine-binding protein 1, PEBP1)；荧光定量PCR验证与双向电泳结果一致。PEBP1蛋白的生物信息学分析表明其性质稳定，不存在信号肽，定位于细胞质，可能属于非分泌性蛋白，含磷脂酰乙醇胺结合蛋白家族信号位点64YTLVLTDPDAPSRKDPKFREWHH86；主要二级结构元件为无规则卷曲和延伸链。同源性分析表明，大鼠PEBP1蛋白与其他9个物种有较高同源性，并构建了PEBP1蛋白的系统进化树。中毒组大鼠肝脏PEBP1蛋白表达下调，导致肝细胞线粒体功能障碍，可能是纳米铜发挥毒性作用的途径之一。
Abstract：In order to screen the differentially expressed protein and explore the mechanism of hepatotoxicity induced by copper nanoparticles in rat(Rattus norvegicus), the differential proteins were separated and identified by two dimensional gel electro-phoresis (2-DE) and MALDI-TOF-TOF MS, and then verified by Real-time PCR, and analyzed by bioinformatics. Total 43 spots of differentially expressed proteins were found，of which a spot 1006 associated with hepatotoxity was identified as phosphatidy lethanolamine-binding protein 1(PEBP1)(accession No. IP1002309), the results of Real-time PCR was the same as that of 2-DE. Bioinformatics analysis showed that PEBP1 was located in the cytoplasm, and had no signal peptide, it may belongs to non-secretory protein, and contain phosphatidy lethanolamine-binding protein family signature 64YTLVLTDPDAPSRKDPKFREWHH86; the random coils and extended chains were its main secondary structural elements. The homology analysis showed that PEBP1 had a high homology between rat and other nine species, and the phylogenetic tree of PEBP1 was constructed by Neighbor-joint method. The expression of PEBP1 protein was reduced in poisoned rats so as to liver cell mitochondrial dysfunction, which may be a pathway of copper nanoparticles to exert the hepatotoxic effects.
