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基于iTRAQ的高压二氧化碳诱导的亚致死损伤O157:H7细胞蛋白质组学分析

iTRAQ-Based Proteomic Analysis of Sublethally Injured O157:H7 Cells Induced by High Pressure Carbon Dioxide.

作者信息

Bi Xiufang, Wang Yongtao, Hu Xiaosong, Liao Xiaojun

机构信息

Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.

Key Lab of Fruit and Vegetable Processing, Ministry of Agriculture, Beijing, China.

出版信息

Front Microbiol. 2017 Dec 18;8:2544. doi: 10.3389/fmicb.2017.02544. eCollection 2017.

Abstract

High pressure carbon dioxide (HPCD) could cause sublethally injured cells (SICs), which may cause food poisoning and spoilage during food storage and limit its application. Therefore, the formation of SICs of O157:H7 was investigated by isobaric tag for relative and absolute quantification (iTRAQ) proteomic methods in this study for better controlling the SICs induced by HPCD. A total of 2,446 proteins was identified by iTRAQ, of which 93 and 29 were significantly differentially expressed in the SICs compared with live control cells (CK) and dead control cells (CK), respectively. Among the 93 differentially expressed proteins (DEP) in the SICs compared with CK, 65 proteins showed down-regulation and 28 showed up-regulation. According to the comprehensive proteome coverage analysis, the SICs survived under HPCD by reducing carbohydrate decomposing, lipid transport and metabolism, amino acid transport and metabolism, transcription and translation, DNA replication and repair. Besides, the SICs showed stress response, DNA damage response and an increased carbohydrate transport, peptidoglycan synthesis and disulfide bond formation to HPCD. Among the 29 DEP in the SICs compared with CK, 12 proteins showed down-regulation and 17 showed up-regulation. According to the comprehensive proteome coverage analysis, the SICs survived under HPCD by accumulation of cell protective agents like carbohydrates and amino acids, and decreasing transcription and translation activities. Results showed that the formation of the SICs with low metabolic activity and high survival ability was a survival strategy for O157:H7 against HPCD.

摘要

高压二氧化碳(HPCD)可导致亚致死损伤细胞(SICs),这可能在食品储存期间引发食物中毒和变质,并限制其应用。因此,本研究采用等压标签相对和绝对定量(iTRAQ)蛋白质组学方法研究了O157:H7的SICs形成情况,以便更好地控制HPCD诱导的SICs。通过iTRAQ共鉴定出2446种蛋白质,其中与活对照细胞(CK)和死对照细胞(CK)相比,分别有93种和29种在SICs中显著差异表达。与CK相比,SICs中93种差异表达蛋白(DEP)中,65种蛋白表达下调,28种蛋白表达上调。根据综合蛋白质组覆盖分析,SICs在HPCD下通过减少碳水化合物分解、脂质转运和代谢、氨基酸转运和代谢、转录和翻译、DNA复制和修复而存活。此外,SICs对HPCD表现出应激反应、DNA损伤反应以及碳水化合物转运、肽聚糖合成和二硫键形成增加。与CK相比,SICs中29种DEP中,12种蛋白表达下调,17种蛋白表达上调。根据综合蛋白质组覆盖分析,SICs在HPCD下通过积累碳水化合物和氨基酸等细胞保护剂以及降低转录和翻译活性而存活。结果表明,具有低代谢活性和高存活能力的SICs的形成是O157:H7对抗HPCD的一种存活策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f22/5770692/3ed2154dff83/fmicb-08-02544-g0001.jpg

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