Wongtrakoongate Patompon, Mongkoldhumrongkul Napachanok, Chaijan Suthidarak, Kamchonwongpaisan Sumalee, Tungpradabkul Sumalee
Department of Biology, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok 10400, Thailand.
Mol Cell Probes. 2007 Apr;21(2):81-91. doi: 10.1016/j.mcp.2006.08.006. Epub 2006 Sep 3.
Burkholderia pseudomallei is a bacterial pathogen causing the melioidosis disease, which is predominantly found in tropical areas of Southeast Asia and Northern Australia. Burkholderia thailandensis is a closely related species to B. pseudomallei but it is non-pathogenic species. In this study, we have constructed a proteome reference map of B. pseudomallei at the stationary phase of growth by using two-dimensional gel electrophoresis with a pH 4-7 immobilized pH gradient combined with matrix-assisted laser desorption ionization time of flight mass spectrometry. Approximately 550 spots could be detected by Coomassie brilliant blue G-250 staining, and 88 spots representing 77 unique proteins were identified. Eleven of the gene products were found in multiple spots indicating as isoforms. In attempt to detect distinctive expressed proteins between a virulent and a non-virulent species, the use of comparative proteomic profiles under the same condition were performed. We could identify more than 20 different spots. Twelve out of 14 spots are detected in B. pseudomallei and six proteins have been identified and indicated that they are involved in virulent characters of bacteria. Two hypothetical proteins were expressed and found only in B. pseudomallei. These proteins are potential markers to distinguish between these two species. Our study also provides a useful information of global intracellular protein expression and is a valuable starting point for analyzing a proteomic pathogenicity of the bacterial pathogen.
类鼻疽伯克霍尔德菌是一种引起类鼻疽病的细菌病原体,主要发现于东南亚和澳大利亚北部的热带地区。泰国伯克霍尔德菌是与类鼻疽伯克霍尔德菌密切相关的物种,但它是无致病力的物种。在本研究中,我们通过使用pH 4 - 7固定化pH梯度的二维凝胶电泳结合基质辅助激光解吸电离飞行时间质谱,构建了类鼻疽伯克霍尔德菌生长稳定期的蛋白质组参考图谱。考马斯亮蓝G - 250染色可检测到约550个斑点,鉴定出代表77种独特蛋白质的88个斑点。其中11种基因产物存在于多个斑点中,表明为同工型。为了检测有毒力和无毒力物种之间差异表达的蛋白质,我们在相同条件下进行了比较蛋白质组分析。我们能够鉴定出20多个不同的斑点。在类鼻疽伯克霍尔德菌中检测到14个斑点中的12个,已鉴定出6种蛋白质,表明它们与细菌的毒力特征有关。两种假定蛋白质仅在类鼻疽伯克霍尔德菌中表达并被发现。这些蛋白质是区分这两个物种的潜在标志物。我们的研究还提供了关于全球细胞内蛋白质表达的有用信息,是分析该细菌病原体蛋白质组致病性的宝贵起点。