付胜利, 刘海粟, 廖绍安, 郭政. 嗜水气单胞菌感染暗纹东方鲀脾脏的蛋白质组学分析[J]. 华南师范大学学报(自然科学版), 2020, 52(5): 76-84. doi: 10.6054/j.jscnun.2020080
引用本文: 付胜利, 刘海粟, 廖绍安, 郭政. 嗜水气单胞菌感染暗纹东方鲀脾脏的蛋白质组学分析[J]. 华南师范大学学报(自然科学版), 2020, 52(5): 76-84. doi: 10.6054/j.jscnun.2020080
FU Shengli, LIU Haisu, LIAO Shaoan, GUO Zheng. A Proteomic Analysis of Pufferfish (Takifugu obscurus) Spleen Under Aeromonas hydrophila Infection[J]. Journal of South China Normal University (Natural Science Edition), 2020, 52(5): 76-84. doi: 10.6054/j.jscnun.2020080
Citation: FU Shengli, LIU Haisu, LIAO Shaoan, GUO Zheng. A Proteomic Analysis of Pufferfish (Takifugu obscurus) Spleen Under Aeromonas hydrophila Infection[J]. Journal of South China Normal University (Natural Science Edition), 2020, 52(5): 76-84. doi: 10.6054/j.jscnun.2020080

嗜水气单胞菌感染暗纹东方鲀脾脏的蛋白质组学分析

A Proteomic Analysis of Pufferfish (Takifugu obscurus) Spleen Under Aeromonas hydrophila Infection

  • 摘要: 为了解暗纹东方鲀(Takifugu obscurus)受到病原菌感染后参与应答的主要蛋白的功能与调控作用,采用TMT标记、高效液相色谱(HPLC)分级技术以及基于质谱的定量蛋白质组学技术,分析了暗纹东方鲀感染嗜水气单胞菌(Aeromonas hydrophila)12 h后脾脏组织的蛋白表达水平,鉴定差异表达蛋白,并通过亚细胞定位、GO(Gene Oncology)和KEGG(Kyoto Encyclopedia of Genes and Genomes)对差异表达蛋白进行进一步分析.本次实验共鉴定到306个差异表达蛋白,其中120个上调表达蛋白,186个下调表达蛋白;利用平行反应检测实验(PRM)对4个(1个上调,3个下调)与免疫相关的差异蛋白进行验证,其结果与蛋白质测序结果一致;亚细胞定位结果显示:差异表达蛋白主要定位在细胞质、细胞核和胞外;GO分析发现:差异表达蛋白参与的主要生物进程有代谢过程、细胞进程和单一生物进程,主要的分子功能有结合、催化活性和机构分析活性;KEGG通路分析结果显示:差异表达蛋白主要参与了核糖体、视黄醇代谢、卟啉和叶绿素代谢、ECM受体相互作用、吞噬体、叶酸生物合成、药物代谢、酪氨酸代谢等通路.研究结果为进一步探究病原菌感染暗纹东方鲀的致病机理提供了一定的理论基础.

     

    Abstract: TMT labeling, high performance liquid chromatography (HPLC) and quantitative proteomics based on mass spectrometry were used to investigate the differential expression proteins (DEPs) in spleen of pufferfish (Takifugu obscurus) infected with Aeromonas hydrophila for 12 h. The spleens of the experimental group and the control group were collected at 12 h after infection, then the expression level of proteins was detected and DEPs were screened through identification and quantification analysis. In addition, DEPs were further analyzed with subcellular localization, gene oncology(GO) and kyoto encyclopedia of genes and genomes(KEGG). A total of 306 DEPs were screened, among which 120 proteins were up-regulated and 186 proteins were down-regulated. Four (1 up-regulated, 3 down-regulated) immune-related proteins were chosen for validation with parallel response assay (PRM), and the results were consistent with those of the quantitative proteomics. The results of subcellular localization showed that DEPs were mainly located in cytoplasm, nucleus and extracellular. GO analysis found that the main biological processes involved in DEPs were metabiolic processes, cellular processes and single biological processes, and the main molecular functions were binding, catalytic activity and mechanism analysis activity. Bioinformatics analysis indicated that DEPs were mainly associated with ribosome, retinol metabolism, porphyrin and chlorophyll metabolism, ECM-receptor interaction, phagosome, folate biosynthesis, drug metabolism, tyrosine metabolism and so on. The potential mechanism of regulated proteins by A. hydrophila infection in spleen pathology needs further investigation.

     

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