School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, Australia.
Mark Wainwright Analytical Centre, University of New South Wales, Sydney, New South Wales, Australia.
Proteomics. 2018 Apr;18(8):e1700237. doi: 10.1002/pmic.201700237.
Bordetella pertussis causes whooping cough. The predominant strains in Australia changed to single nucleotide polymorphism (SNP) cluster I (pertussis toxin promoter allele ptxP3/pertactin gene allele prn2) from cluster II (non-ptxP3/non-prn2). Cluster I was mostly responsible for the 2008-2012 Australian epidemic and was found to have higher fitness compared to cluster II using an in vivo mouse competition assay, regardless of host's immunization status. This study aimed to identify proteomic differences that explain higher fitness in cluster I using isobaric tags for relative and absolute quantification (iTRAQ), and high-resolution multiple reaction monitoring (MRM-hr). A few key differences in the whole cell and secretome were identified between the cluster I and II strains tested. In the whole cell, nine proteins were upregulated (>1.2 fold change, q < 0.05) and three were downregulated (<0.8 fold change, q < 0.05) in cluster I. One downregulated protein was BP1569, a TLR2 agonist for Th1 immunity. In the secretome, 12 proteins were upregulated and 1 was downregulated which was Bsp22, a type III secretion system (T3SS) protein. Furthermore, there was a trend of downregulation in three T3SS effectors and other virulence factors. Three proteins were upregulated in both whole cell and supernatant: BP0200, molybdate ABC transporter (ModB), and tracheal colonization factor A (TcfA). Important expression differences in lipoprotein, T3SS, and transport proteins between the cluster I and II strains were identified. These differences may affect immune evasion, virulence and metabolism, and play a role in increased fitness of cluster I.
百日咳鲍特菌引起百日咳。澳大利亚的主要菌株已从 II 群（非 ptxP3/非 prn2）转变为单核苷酸多态性（SNP）I 群（百日咳毒素启动子等位基因 ptxP3/pertactin 基因等位基因 prn2）。I 群主要负责 2008-2012 年澳大利亚的流行，并在体内小鼠竞争试验中被发现比 II 群具有更高的适应性，而与宿主的免疫状态无关。本研究旨在使用相对和绝对定量同位素标记（iTRAQ）和高分辨率多重反应监测（MRM-hr）来鉴定解释 I 群更高适应性的蛋白质组学差异。在整个细胞和分泌组中，I 群和 II 群菌株之间鉴定出几个关键差异。在整个细胞中，有 9 种蛋白上调（>1.2 倍变化，q < 0.05），3 种蛋白下调（<0.8 倍变化，q < 0.05）。下调蛋白之一是 BP1569，一种 TLR2 激动剂，可诱导 Th1 免疫。在分泌组中，上调了 12 种蛋白，下调了 1 种蛋白，即 Bsp22，一种 III 型分泌系统（T3SS）蛋白。此外，还有三种 T3SS 效应子和其他毒力因子呈下调趋势。三种蛋白在整个细胞和上清液中均上调：BP0200、钼 ABC 转运蛋白（ModB）和气管定植因子 A（TcfA）。鉴定了 I 群和 II 群菌株之间脂蛋白、T3SS 和转运蛋白的重要表达差异。这些差异可能影响免疫逃避、毒力和代谢，并在 I 群适应性增加中发挥作用。
