Proteomic analysis of differentially expressed proteins in Penaeus vannamei hemocytes upon Taura syndrome virus infection
Proteomic analysis of differentially expressed proteins in Penaeus vannamei hemocytes upon Taura syndrome virus infection
复制标题
DOI：
10.1002/pmic.200700281
复制
发表时间：
2007-10-01
期刊：
影响因子：
3.4
通讯作者：
Krittanai, Chartchai
中科院分区：
文献类型：
--
作者：
Chongsatja, Phattara-orn;Bourchookarn, Apichai;Krittanai, Chartchai
为了了解甲壳动物血细胞对病毒感染的分子反应，我们应用双向电泳蛋白质组学技术研究了南美白对虾（Penaeus vannamei）Taura感染期间血细胞蛋白质的变化。综合征病毒（TSV）感染。在感染后24 h，定量强度分析和nano-LC-ESI-MS/MS显示8种蛋白质中的11种形式显著上调，而5种蛋白质中的9种形式显著下调。这些改变的蛋白质在宿主防御（血蓝蛋白、过氧化氢酶、羧酸酯酶、转谷氨酰胺酶和谷胱甘肽转移酶）、信号传导（14-3-3 zeta）、碳水化合物代谢（乙酰葡萄糖胺焦磷酸化酶）、细胞结构和完整性（β-微管蛋白、β-肌动蛋白、原肌球蛋白和肌球蛋白）以及ER应激反应（蛋白质二硫键异构酶）中发挥重要作用。半定量RT-PCR和Western blot分析证实了14-3-3在mRNA和蛋白水平上的上调。有趣的是，几个改变的蛋白质点被鉴定为血蓝蛋白的片段。质谱分析表明，在酸性和碱性区域的血蓝蛋白斑点分别代表C-和N-末端血蓝蛋白片段。由于四分之三的C-末端片段被上调，而三分之二的N-末端血蓝蛋白片段被下调，因此我们假设C-和N-末端血蓝蛋白片段可能在血细胞中具有不同的作用。这些数据的进一步调查可能会导致更好地了解甲壳动物血细胞TSV感染的分子反应。
To understand molecular responses of crustacean hemocytes to virus infection, we applied 2-DE proteomics approach to investigate altered proteins in hemocytes of Penaeus vannamei during Taura. syndrome virus (TSV) infection. At 24 h postinfection, quantitative intensity analysis and nano-LC-ESI-MS/MS revealed 11 forms of 8 proteins that were significantly up-regulated, whereas 9 forms of 5 proteins were significantly down-regulated in the infected shrimps. These altered proteins play important roles in host defense (hemocyanin, catalase, carboxylesterase, transglutaminase, and glutathione tyansferase), signal transduction (14-3-3 zeta), carbohydrate metabolism (acetylglucosamine pyrophosphorylase), cellular structure and integrity (beta-tubulin, beta-actin, tropomyosin, and myosin), and ER-stress response (protein disulfide isomerase). Semiquantitative RT-PCR and Western blot analysis confirmed the upregulation of 14-3-3 at both mRNA and protein levels. Interestingly, several altered protein spots were identified as fragments of hemocyanin. Mass spectrometric analysis showed that the hemocyanin spots at acidic and basic regions represented the C- and N-terminal hemocyanin fragments, respectively. As three-quarters of C-terminal fragments were up-regulated, whereas two-thirds of N-terminal hemocyanin fragments were down-regulated, we therefore hypothesize that C- and N-terminal hemocyanin fragments may have differential roles in hemocytes. Further investigation of these data may lead to better understanding of the molecular responses of crustacean hemocytes to TSV infection.