Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, PR China.
J Proteomics. 2012 Jan 4;75(3):894-907. doi: 10.1016/j.jprot.2011.10.004. Epub 2011 Oct 25.
In this report, we carried out the in-depth proteomic analysis of Yersinia pestis strain 91001 under in vitro flea-simulated condition using three technique routes, SDS-PAGE combined with LTQ-FT, two-dimensional liquid chromatography peptide (2D-LC peptide) separation combined with LTQ-FT and intact protein separation followed by 2D-LC peptide separation combined with LTQ-FT. Totally, 1926 proteins (13082 peptides) were identified, covering 46.50% (1926/4142) of the predicted proteome. Transcriptome analysis based on a whole genome DNA microarray of Y. pestis defined 1655 genes with the coincidence of 56.65% to the proteomic results. Through analyzing the identifications of virulence factors involving in the life cycle of Y. pestis, it was found that Hms system and murine toxin, which are virulence factors involved in Y. pestis maintenance in flea, were highly expressed in our analysis. Moreover, some virulence factors also appeared with different extents, such as plasminogen activator, PhoP/PhoQ two-component system, type III secretion system, iron acquisition systems (Ybt, Yfe and Yfu) and ferric uptake regulator. These results indicated that Y. pestis may prepare itself with various strategies in advance for its survival when it evades the hosts. The protein identifications can be accessed through PRIDE database (http://www.ebi.ac.uk/pride) with accession number 18578-18605
在本报告中,我们使用三种技术路线(SDS-PAGE 联合 LTQ-FT、二维液相色谱肽(2D-LC peptide)分离联合 LTQ-FT 和完整蛋白质分离后联合 2D-LC peptide 分离)对体外跳蚤模拟条件下的鼠疫耶尔森氏菌 91001 进行了深入的蛋白质组学分析。总共鉴定了 1926 种蛋白质(13082 种肽),覆盖了预测蛋白质组的 46.50%(1926/4142)。基于鼠疫耶尔森氏菌全基因组 DNA 微阵列的转录组分析,有 1655 个基因与蛋白质组学结果的符合度为 56.65%。通过分析涉及鼠疫耶尔森氏菌生命周期的毒力因子的鉴定,发现 Hms 系统和鼠毒素是鼠疫耶尔森氏菌在跳蚤中维持的毒力因子,在我们的分析中表达水平较高。此外,一些毒力因子也表现出不同程度的出现,如纤溶酶原激活物、PhoP/PhoQ 双组分系统、III 型分泌系统、铁获取系统(Ybt、Yfe 和 Yfu)和铁摄取调节剂。这些结果表明,鼠疫耶尔森氏菌在逃避宿主时可能会提前准备各种策略来维持自身的生存。蛋白质鉴定可通过 PRIDE 数据库(http://www.ebi.ac.uk/pride)访问,访问号为 18578-18605。