[1]赵为民,涂枫,王丽,等.猪PKM2基因在肺炎支原体感染3D4/21细胞后的互作蛋白质鉴定与分析[J].江苏农业学报,2019,(06):1381-1389.[doi:doi:10.3969/j.issn.1000-4440.2019.06.016]
　ZHAO Wei-min,TU Feng,WANG Li,et al.Identification and analysis of interaction proteins of porcine PKM2 gene in 3D4/21 cells during Mycoplasma pneumoniae infection[J].,2019,(06):1381-1389.[doi:doi:10.3969/j.issn.1000-4440.2019.06.016]
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猪PKM2基因在肺炎支原体感染3D4/21细胞后的互作蛋白质鉴定与分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年06期
页码:
1381-1389
栏目:
畜牧兽医·水产养殖
出版日期:
2019-12-31

文章信息/Info

Title:
Identification and analysis of interaction proteins of porcine PKM2 gene in 3D4/21 cells during Mycoplasma pneumoniae infection
作者:
赵为民1; 2; 涂枫1; 2; 王丽1; 2; 任守文1; 2; 付言峰1; 2; 李碧侠1; 2; 徐小波1; 2; 陈哲1; 2; 方晓敏1; 2
（1. 江苏省农业科学院畜牧研究所/江苏省农业种质资源保护与利用平台，江苏南京210014； 2. 农业部种养结合重点实验室，江苏南京210014）
Author(s):
ZHAO Wei-min1; 2; TU Feng1; 2; WANG Li1; 2; REN Shou-wen1; 2; FU Yan-feng1; 2; LI Bi-xia1; 2; XU Xiao-bo1; 2; CHEN Zhe1; 2; FANG Xiao-min1; 2
(1.Institute of Animal Science, Jiangsu Academy of Agricultural Sciences/Jiangsu Germplasm Resources Protection and Utilization Platform, Nanjing 210014, China; 2. Key Laboratory of Crop and Livestock Integration, Ministry of Agriculture, Nanjing 210014, China)
关键词:
猪; PKM2基因; 肺炎支原体; 互作蛋白质; 免疫共沉淀
Keywords:
porcine; PKM2 gene; Mycoplasma pneumoniae; interaction protein; co-immunoprecipitation(Co-IP)
分类号:
S852.62
DOI:
doi:10.3969/j.issn.1000-4440.2019.06.016
文献标志码:
A
摘要:
通过构建含有HA标签的PKM2过表达质粒转染3D4/21细胞，利用免疫共沉淀（Co-IP）与液相色谱-质谱联用（LC-MS/MS）技术分析与鉴定猪PKM2基因在肺炎支原体感染猪肺泡巨噬细胞3D4/21后的互作蛋白质，并对这些互作蛋白质进行GO与KEGG通路分析。结果显示肺炎支原体感染3D4/21细胞后，PKM2在mRNA水平和蛋白质水平上表达上调，其表达水平呈现剂量与时间依懒性。免疫共沉淀反应结果显示IP与IgG组共鉴定到72个蛋白质，这2组鉴定到的蛋白质数分别为60和19，其中7个蛋白质在2组中同时鉴定到。GO分析结果表明IP组的互作蛋白质主要参与了NAD代谢过程、NADH代谢过程、肌原纤维组装、NADH 再生和葡萄糖代谢丙酮酸等生物过程，其中大多数与能量代谢有关。KEGG pathway分析结果表明互作蛋白质参与了致病性大肠杆菌感染、军团杆菌病、糖酵解/糖异生、丙酮酸代谢和癌症中的中枢碳代谢等通路，这些通路与细菌感染和能量代谢有关。
Abstract:
3D4/21 cells were transfected withHA-tagged PKM2-overexpression plasmid, and the co-immunoprecipitation (Co-IP) and liquid chromatography-mass spectrometry (LC-MS/MS) technique were used to identify and analyze the interaction protein of porcine PKM2 gene in porcine alveolar macrophages 3D4/21 during Mycoplasma pneumoniae infection. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotations were performed on these interaction proteins. The results showed that PKM2 was up-regulated at mRNA and protein levels after Mycoplasma pneumoniae infection in 3D4/21, and its expression level was dose and time-dependent. A total of 72 proteins were identified in the IP and IgG groups in the co-immunoprecipitation reaction. The number of proteins identified in the above two groups was 60 and 19, respectively, and seven proteins were identified in both groups. GO analysis for biological processes showed that the interaction proteins of IP group were mainly involved in NAD metabolism process, NADH metabolism process, myofibril assembly, NADH regeneration and glucose catabolic process to pyruvate, most of which were related to energy metabolism. The KEGG pathway results indicated that the interacting proteins were involved in pathogenic Escherichia coli infection, legionellosis, glycolysis/gluconeogenesis, pyruvate metabolism, and central carbon metabolism in cancer, which were related to bacteria infection and energy metabolism.

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备注/Memo

备注/Memo:
收稿日期：2019-03-12 基金项目：国家自然科学基金项目（31601928）；江苏省农业重大新品种创制项目(PZCZ201733)；江苏省农业科学院基本科研业务专项[ZX(15)4002] 作者简介：赵为民（1983-），男，湖北钟祥人，博士，副研究员，主要从事猪的抗病育种， (E-mail)zhao_weimin1983@aliyun.com 通讯作者：方晓敏，(E-mail)fxmw2000@163.com
更新日期/Last Update: 2020-01-09