A quantitative proteomic screen of the Campylobacter jejuni flagellar-dependent secretome
A quantitative proteomic screen of the Campylobacter jejuni flagellar-dependent secretome
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DOI：
10.1016/j.jprot.2016.11.009
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发表时间：
2017-01-30
影响因子：
3.3
通讯作者：
Grant, Andrew
中科院分区：
文献类型：
--
作者：
Scanlan, Eoin;Yu, Lu;Grant, Andrew
空肠弯曲菌是世界上细菌性胃肠炎的主要原因。据信，许多因素导致空肠弯曲菌在人类宿主内引起疾病，包括通过鞭毛 III 型分泌系统 (FT3SS) 分泌非鞭毛蛋白。在这里，我们首次利用细胞培养物中氨基酸稳定同位素标记的定量蛋白质组学 (SILAC) 和无标记液相色谱-质谱 (LC/MS) 来比较来自空肠弯曲菌 M1 野生型和鞭毛缺陷（flgG 突变体）菌株的上清液样品，以鉴定通过 FT3SS 分泌的假定新蛋白质。来自 LC/MS 和 SILAC 数据集的编码鞭毛分泌候选蛋白的基因被删除。使用这些突变体感染人类 CACO-2 组织培养细胞，导致鉴定出与这些细胞相互作用所需的新基因。这项工作首次表明，CJM1_0791 和 CJM1_0395 在空肠弯曲菌染色 M1 和 81-176 上清液中的存在都依赖于鞭毛。 生物学意义：这项研究提供了迄今为止对空肠弯曲杆菌分泌组的最完整的描述。 SILAC 和无标记蛋白质组学比较了有或没有鞭毛的突变体，结果鉴定出两种空肠弯曲菌蛋白依赖于鞭毛从细菌细胞中输出。 (C) 2016 年作者。由 Elsevier B.V. 出版
Campylobacter jejuni is the leading cause of bacterial gastroenteritis in the world. A number of factors are believed to contribute to the ability of C. jejuni to cause disease within the human host including the secretion of non-flagellar proteins via the flagellar type III secretion system (FT3SS). Here for the first time we have utilised quantitative proteomics using stable isotope labelling by amino acids in cell culture (SILAC), and label-free liquid chromatography-mass spectrometry (LC/MS), to compare supernatant samples from C. jejuni M1 wild type and flagella-deficient (flgG mutant) strains to identify putative novel proteins secreted via the FT3SS. Genes encoding proteins that were candidates for flagellar secretion, derived from the LC/MS and SILAC datasets, were deleted. Infection of human CACO-2 tissue culture cells using these mutants resulted in the identification of novel genes required for interactions with these cells. This work has shown for the first time that both CJM1_0791 and CJM1_0395 are dependent on the flagellum for their presence in supernatants from C jejuni stains M1 and 81-176.Biological significance: This study provides the most complete description of the Campylobac er jejuni secretome to date. SILAC and label-free proteomics comparing mutants with or without flagella have resulted in the identification of two C. jejuni proteins that are dependent on flagella for their export from the bacterial cell. (C) 2016 The Authors. Published by Elsevier B.V.