布鲁菌16M感染后宿主免疫相关蛋白质泛素化修饰的差异分析

doi:10.11843/j.issn.0366-6964.2019.11.013

摘要/Abstract

摘要： 为探究布鲁菌感染宿主细胞早期泛素化修饰蛋白质组表达变化并筛选出影响免疫应答的关键调控蛋白，通过Label-free和泛素化富集技术以及高分辨率LC-MS/MS联用的定量蛋白质组学研究策略，对布鲁菌16M感染11 h（感染5 h，胞内复制6 h）后的巨噬细胞和未感染布鲁菌16M的巨噬细胞进行了泛素化蛋白质组学定量研究，数据库检索分析了16M感染后的巨噬细胞和未感染的巨噬细胞差异表达的泛素化位点对应的蛋白质，并应用生物信息学方法筛选出16M感染巨噬细胞后可造成宿主免疫抑制的关键蛋白质。结果显示，共鉴定出349个蛋白质上580个泛素化位点。布鲁菌16M感染组相对于未感染组167个蛋白质上259个位点泛素化修饰水平发生上调，212个蛋白质上321个位点泛素化修饰水平发生下调（差异倍数>1.5，P<0.05）；35个泛素化修饰差异表达的蛋白质可能与布鲁菌感染后宿主的免疫应答有关，其中27个泛素化修饰的下调蛋白质（如Bcap31、Btk、Faf1和Akap31等），1个泛素化修饰上调蛋白质Ubqln1可能是布鲁菌感染宿主后造成免疫抑制的关键蛋白。本研究筛选获得了布鲁菌16M感染宿主细胞免疫相关差异表达的泛素化修饰蛋白质，初步揭示布鲁菌能够调控宿主免疫信号通路、自噬和凋亡等免疫应答过程中相关蛋白质的泛素化修饰，为进一步研究布鲁菌感染后调节宿主泛素化修饰进而形成免疫逃逸的分子机制提供理论基础。

Abstract: This experiment was conducted to investigate the expression level of ubiquitinated modified proteins in host cells infected with Brucella in early stage and screen out the key regulatory proteins that affect the process of immune response. By using Label-free and ubiquitination enrichment technology and high resolution LC-MS/MS combined quantitative proteomics research strategy, ubiquitination proteomics quantitative studies were carried out on Brucella 16M infected macrophages and uninfected macrophages after 11 hours (5 hours' infection, 6 hours' intracellular replication). Database retrieval analysis was performed on the proteins which were corresponding to the ubiquitination sites that differentially expressed by 16M infected macrophages and uninfected macrophages. The key proteins that can cause host immunosuppression after 16M infection of macrophages were screened out by bioinformatics method. In this study, 580 ubiquitination sites were identified on 349 proteins. The ubiquitination level of 259 sites on 167 proteins in the 16M infection group was up-regulated compared with the uninfected group, 321 sites on 212 proteins were down-regulated (difference multiple >1.5, P<0.05); There were 35 ubiquitination-modified differentially expressed proteins may be related to the host immune response after Brucella infection. Among them, we found 27 ubiquitinated down-regulated proteins such as Bcap31, Btk, Faf1 and Akap31; one up-regulated ubiquitinated protein, Ubqln1, may be the key protein causing immunosuppression after Brucella infection. In this study, ubiquitinated modified proteins differentially expressed in host cell immune response to Brucella 16M infection were screened and obtained. It was preliminarily revealed that Brucella can affect the ubiquitinated modification of related proteins in host immune signaling pathway, autophagy and apoptosis, so as to further study the regulation of perennial status after Brucella infection. The main ubiquitination modification to complete the molecular mechanism of immune escape provides a theoretical basis.

中图分类号:

S852.614

周玉成, 郭梦楠, 程世鹏, 张海威, 周曼莉, 乔连江, 杨艳玲. 布鲁菌16M感染后宿主免疫相关蛋白质泛素化修饰的差异分析[J]. 畜牧兽医学报, 2019, 50(11): 2290-2301.

ZHOU Yucheng, GUO Mengnan, CHENG Shipeng, ZHANG Haiwei, ZHOU Manli, QIAO Lianjiang, YANG Yanling. Differential Analysis of Ubiquitination Modification of Host Immune-related Proteins after Brucella 16M Infection[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(11): 2290-2301.

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