基于差异蛋白质组学分析氧化应激下耐辐射奇球菌的抗氧化机制  

Antioxidative mechanism of Deinococcus radiodurans under oxidative stress based on differential proteomics

作　　者：王馥丽 魏琳洋 甘巧玉 蒋辉 宋天宇 姚戈 万秀坤 Wang Fuli;Wei Linyang;Gan Qiaoyu;Jiang Hui;Song Tianyu;Yao Ge;Wan Xiukun(State Key Laboratory of National Nuclear,Biological and Chemical Disaster Protection,Beijing 102205;School of Life,Beijing Institute of Technology,No.5 Zhongguancun South Street,Beijing 100081)

机构地区：[1]国民核生化灾害防护国家重点实验室,北京102205 [2]北京市中关村南大街5号北京理工大学生命学院,北京100081

出　　处：《生命科学仪器》2023年第3期54-63,共10页Life Science Instruments

基　　金：国民核生化灾害防护国家重点实验室科研基金项目(SKLNBC2020-13)

摘　　要：耐辐射奇球菌具有极强的生命力,被吉尼斯世界记录收录并誉为是“世界上最顽强的细菌”,也作为模式生物用于抗性机制研究,电离辐射、强氧化剂等物理化学作用导致细胞产生大量具有高度细胞毒性作用的活性氧自由基,从而间接对生命体产生氧化损伤,对ROS带来氧化损伤其也具有极强的抗氧化机制,对但其抗氧化机制的研究尚未完全。本研究将采用非标记定量蛋白质组学分析氧化应激下耐辐射奇球菌的抗氧化机制,基于非标记定量蛋白质组学分析,H2O2处理后的耐辐射奇球菌共检测到1853个蛋白,其中80个蛋白表达量与处理前相比具有显著差异,其中25个显著上调,55个显著下调。4h恢复培养后,耐辐射奇球菌组共检测到1853个蛋白,其中58个蛋白表达量发生显著变化,38个显著上调,20个显著下调。基于差异蛋白进行生物信息学分析并比较差异蛋白类型,我们发现H2O2氧化处理产生的差异蛋白主要富集在抵抗氧化应激胁迫等生物学过程;恢复培养4h产生的差异蛋白在恢复耐辐射奇球菌的正常生理代谢生物过程中显著富集。分析可知,在强氧化剂作用状态下,耐辐射奇球菌产生保护机制,翻译、核糖体结构相关蛋白上调以的合成更多抵御氧化损伤的蛋白,恢复培养后,重建恢复平衡相关蛋白的上调使得耐辐射奇球菌能够更快捷恢复正常生长状态。Deinococcus radiodurans has the strong vitality.It is included in the Guinness World Records and praised as"the most tenacious bacteria in the world".It is also used as a model organism for the study of resistance mechanism.Physical and chemical effects such as ionizing radiation and strong oxidants lead to the production of a large number of reactive oxygen free radicals with highly cytotoxic effects,which indirectly cause oxidative damage to life.It brings oxidative damage to ROS,and Deinococcus radiodurans also has a strong antioxidant mechanism,but the research on its antioxidant mechanism has not been completed.In this study,differential proteomics will be used to analyze the antioxidant mechanism of Deinococcus radiodurans under oxidative stress.Based on label-free,1853 proteins were detected in Deinococcus radiodurans after H_(2)O_(2)treatment,of which 80 proteins had significant differences compared with those before treatment,of which 25 were significantly up-regulated and 55 were significantly down regulated.After 4 h of culture,1853 proteins were detected in Deinococcus radiodurans group,of which 58 proteins were significantly changed,38 were significantly up-regulated and 20 were significantly down regulated.Based on the bioinformatics analysis of differential proteins and comparing the types of differential proteins,we found that the differential proteins produced by H_(2)O_(2)oxidation were mainly enriched in biological processes such as resistance to oxidative stress.The differential proteins produced after 4 hours of culture recovery were significantly enriched in the process of restoring the normal physiological metabolism of Deinococcus radiodurans.The analysis shows that under the action of strong oxidants,Deinococcus radiodurans produces a protective mechanism,and the translation and ribosomal structure related proteins are up-regulated to synthesize more proteins that resist oxidative damage.After culture is restored,the balance is restored,and the up regulation of related proteins enables De

关 键 词：耐辐射奇球菌 氧化应激 抗氧化机制 蛋白质组学 非标记定量分析 

分 类 号：R378.1[医药卫生—病原生物学]