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Quantitative Proteomic Analysis of Porcine Intestinal Epithelial Cells Infected with Porcine Deltacoronavirus Using iTRAQ-Coupled LC-MS/MS
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-10-12 , DOI: 10.1021/acs.jproteome.0c00592 Xinrong Zhou 1 , Lei Zhou 1 , Xinna Ge 1 , Xin Guo 1 , Jun Han 1 , Yongning Zhang 1 , Hanchun Yang 1
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-10-12 , DOI: 10.1021/acs.jproteome.0c00592 Xinrong Zhou 1 , Lei Zhou 1 , Xinna Ge 1 , Xin Guo 1 , Jun Han 1 , Yongning Zhang 1 , Hanchun Yang 1
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Porcine deltacoronavirus (PDCoV) is an emergent enteropathogenic coronavirus associated with swine diarrhea. Porcine small intestinal epithelial cells (IPEC) are the primary target cells of PDCoV infection in vivo. Here, isobaric tags for relative and absolute quantification (iTRAQ) labeling coupled to liquid chromatography–tandem mass spectrometry (LC-MS/MS) was used to quantitatively identify differentially expressed proteins (DEPs) in PDCoV-infected IPEC-J2 cells. A total of 78 DEPs, including 23 upregulated and 55 downregulated proteins, were identified at 24 h postinfection. The data are available via ProteomeXchange with identifier PXD019975. To ensure reliability of the proteomics data, two randomly selected DEPs, the downregulated anaphase-promoting complex subunit 7 (ANAPC7) and upregulated interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), were verified by real-time PCR and Western blot, and the results of which indicate that the proteomics data were reliable and valid. Bioinformatics analyses, including GO, COG, KEGG, and STRING, further demonstrated that a majority of the DEPs are involved in numerous crucial biological processes and signaling pathways, such as immune system, digestive system, signal transduction, RIG-I-like receptor, mTOR, PI3K-AKT, autophagy, and cell cycle signaling pathways. Altogether, this is the first study on proteomes of PDCoV-infected host cells, which shall provide valuable clues for further investigation of PDCoV pathogenesis.
中文翻译：
使用 iTRAQ 耦合 LC-MS/MS 对感染猪 Delta 冠状病毒的猪肠上皮细胞进行定量蛋白质组学分析
猪三角洲冠状病毒（PDCoV）是一种与猪腹泻相关的新出现的肠道致病性冠状病毒。猪小肠上皮细胞（IPEC）是体内PDCoV感染的主要靶细胞。在这里，使用用于相对和绝对定量（iTRAQ）标记的同量异位标签与液相色谱-串联质谱（LC-MS/MS）相结合来定量鉴定PDCoV感染的IPEC-J2细胞中的差异表达蛋白（DEP）。感染后 24 小时共鉴定出 78 个 DEP，包括 23 个上调蛋白和 55 个下调蛋白。数据可通过 ProteomeXchange 获取，标识符为 PXD019975。为了确保蛋白质组学数据的可靠性，通过实时PCR和Western blot验证了两个随机选择的DEP，即下调的后期促进复合物亚基7（ANAPC7）和上调的干扰素诱导的四三肽重复蛋白1（IFIT1），并进行了验证。结果表明蛋白质组学数据可靠、有效。包括GO、COG、KEGG和STRING在内的生物信息学分析进一步证明，大多数DEP参与许多关键的生物过程和信号通路，如免疫系统、消化系统、信号转导、RIG-I样受体、 mTOR、PI3K-AKT、自噬和细胞周期信号通路。总之，这是首次对PDCoV感染宿主细胞的蛋白质组进行研究，这将为进一步研究PDCoV发病机制提供有价值的线索。
更新日期：2020-11-06
中文翻译：
使用 iTRAQ 耦合 LC-MS/MS 对感染猪 Delta 冠状病毒的猪肠上皮细胞进行定量蛋白质组学分析
猪三角洲冠状病毒（PDCoV）是一种与猪腹泻相关的新出现的肠道致病性冠状病毒。猪小肠上皮细胞（IPEC）是体内PDCoV感染的主要靶细胞。在这里，使用用于相对和绝对定量（iTRAQ）标记的同量异位标签与液相色谱-串联质谱（LC-MS/MS）相结合来定量鉴定PDCoV感染的IPEC-J2细胞中的差异表达蛋白（DEP）。感染后 24 小时共鉴定出 78 个 DEP，包括 23 个上调蛋白和 55 个下调蛋白。数据可通过 ProteomeXchange 获取，标识符为 PXD019975。为了确保蛋白质组学数据的可靠性，通过实时PCR和Western blot验证了两个随机选择的DEP，即下调的后期促进复合物亚基7（ANAPC7）和上调的干扰素诱导的四三肽重复蛋白1（IFIT1），并进行了验证。结果表明蛋白质组学数据可靠、有效。包括GO、COG、KEGG和STRING在内的生物信息学分析进一步证明，大多数DEP参与许多关键的生物过程和信号通路，如免疫系统、消化系统、信号转导、RIG-I样受体、 mTOR、PI3K-AKT、自噬和细胞周期信号通路。总之，这是首次对PDCoV感染宿主细胞的蛋白质组进行研究，这将为进一步研究PDCoV发病机制提供有价值的线索。
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