Institute of Chemical Biology, Ilia State University , Tbilisi , Georgia ; I.Beritashvili Center for Experimental Biomedicine , Tbilisi , Georgia.
Institute of Chemical Biology, Ilia State University , Tbilisi , Georgia ; National Center for Disease Control , Tbilisi , Georgia.
Front Public Health. 2015 Oct 16;3:239. doi: 10.3389/fpubh.2015.00239. eCollection 2015.
Yersinia pestis, the causative agent of plague, is a highly virulent bacterium responsible for millions of human deaths throughout history. In the last decade, two natural plague foci have been described in the Republic of Georgia from which dozens of Y. pestis strains have been isolated. Analyses indicate that there are genetic differences between these strains, but it is not known if these differences are also reflected in protein expression. We chose four strains of Y. pestis (1390, 1853, 2944, and 8787) from the National Center for Disease Control and Public Health collection for proteomic studies based on neighbor-joining tree genetic analysis and geographical loci of strain origin. Proteomic expression was analyzed using two-dimensional gel electrophoresis and mass spectrometry. Select Y. pestis strains were grown under different physiological conditions and their proteomes were compared: (1) 28°C without calcium; (2) 28°C with calcium; (3) 37°C without calcium; and (4) 37°C with calcium. Candidate proteins were identified and the differences in expression of F1 antigen, tellurium-resistance protein, and outer membrane protein C, porin were validated by Western blotting. The in vitro cytotoxicity activity of these strains was also compared. The results indicate that protein expression and cytotoxic activities differ significantly among the studied strains; these differences could contribute to variations in essential physiological functions in these strains.
鼠疫耶尔森菌是鼠疫的病原体，是一种高度致命的细菌，在历史上导致了数百万人死亡。在过去的十年中，在格鲁吉亚共和国已经描述了两个自然鼠疫疫源地，从中分离出了数十株鼠疫耶尔森菌。分析表明，这些菌株之间存在遗传差异，但尚不清楚这些差异是否也反映在蛋白质表达上。我们根据邻接法遗传分析和菌株起源的地理位置，从国家疾病控制和公共卫生中心收集的四个鼠疫耶尔森菌（1390、1853、2944 和 8787）菌株中选择了用于蛋白质组学研究的四个菌株。使用二维凝胶电泳和质谱法分析蛋白质组表达。选择了一些 Y. pestis 菌株在不同的生理条件下生长，并比较它们的蛋白质组：（1）28°C 无钙；（2）28°C 有钙；（3）37°C 无钙；（4）37°C 有钙。鉴定候选蛋白，并通过 Western blot 验证 F1 抗原、碲抗性蛋白和外膜蛋白 C、孔蛋白的表达差异。还比较了这些菌株的体外细胞毒性活性。结果表明，研究的菌株之间的蛋白质表达和细胞毒性活性存在显著差异；这些差异可能导致这些菌株在基本生理功能上的变化。
