Asakura Hiroshi, Kawamoto Keiko, Murakami Satoshi, Tachibana Masato, Kurazono Hisao, Makino Sou-Ichi, Yamamoto Shigeki, Igimi Shizunobu
Division of Biomedical Food Research, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan.
Research Center for Animal Hygiene and Food Safety, Obihiro University of Agriculture and Veterinary Medicine, Inada 2-11, Obihiro, Hokkaido 080-8555, Japan.
Res Microbiol. 2016 Feb-Mar;167(2):63-71. doi: 10.1016/j.resmic.2015.10.001. Epub 2015 Oct 21.
Campylobacter jejuni is one of the leading causes of foodborne gastrointestinal illness worldwide. Here we performed ex vivo proteomic analysis of C. jejuni 81-176 in chicken, a main reservoir for human infection. At 0, 1 and 4 weeks post-infection (p.i.) with the GFP-expressing 81-176 strain, inocula were recovered from chicken ceca by cell sorting using flow cytometry. iTRAQ-coupled 2D-LC-MS/MS analyses that detected 55 C. jejuni proteins, among which either 3 (FabG, HydB, CJJ81176_0876) or 7 (MscS, CetB, FlhF, PurH, PglJ, LpxC, Icd) proteins exhibited >1.4-fold-increased expression at 1 or 4 week(s) p.i. compared with those at 0 weeks p.i., respectively. Deletion of the fabG gene clearly decreased the proportion of bacterial unsaturated fatty acids (UFAs) and chicken colonization. The UFA proportion of the parental strain was not altered when grown at 42 °C. These findings suggest that FabG might play a pivotal role in UFA production, linked to bacterial adaptation in the poultry host. To our knowledge, this is the first example of ex vivo C. jejuni proteomics, in which fatty acid metabolism might affect bacterial adaptation to the chicken host.
空肠弯曲菌是全球食源性胃肠道疾病的主要病因之一。在此,我们对空肠弯曲菌81-176在鸡(人类感染的主要宿主)体内进行了体外蛋白质组学分析。在用表达绿色荧光蛋白的81-176菌株感染鸡后0、1和4周,通过流式细胞术细胞分选从鸡盲肠中回收接种物。iTRAQ耦合二维液相色谱-串联质谱分析检测到55种空肠弯曲菌蛋白,其中分别有3种(FabG、HydB、CJJ81176_0876)或7种(MscS、CetB、FlhF、PurH、PglJ、LpxC、Icd)蛋白在感染后1周或4周时的表达相较于感染后0周时分别增加了1.4倍以上。fabG基因的缺失明显降低了细菌不饱和脂肪酸(UFA)的比例以及在鸡体内的定殖。亲本菌株在42°C培养时UFA比例未发生改变。这些发现表明,FabG可能在UFA产生中起关键作用,这与细菌在家禽宿主中的适应性有关。据我们所知,这是首次进行的空肠弯曲菌体外蛋白质组学研究实例,其中脂肪酸代谢可能影响细菌对鸡宿主的适应性。