Shotgun proteomic analysis of Chlamydia trachomatis
Shotgun proteomic analysis of Chlamydia trachomatis
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DOI：
10.1002/pmic.200401044
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发表时间：
2005-04-01
期刊：
影响因子：
3.4
通讯作者：
Clarke, IN
中科院分区：
文献类型：
--
作者：
Skipp, P;Robinson, J;Clarke, IN
衣原体是一种广泛存在的细菌病原体，可引起多种疾病，包括性传播感染、肺炎和沙眼。为了验证存在迄今为止假设的蛋白质预测从最近的衣原体基因组测序项目，并检查在蛋白质水平上的关键成分的表达模式，我们已经调查了沙眼衣原体菌株L2的表达蛋白质组。二维凝胶分析，多维蛋白质鉴定（MudPIT）和纳米毛细管液相色谱-串联质谱法的组合允许共328衣原体蛋白质被明确分配。被鉴定为以衣原体的代谢惰性形式（基本体）表达的蛋白质包括整套预测的糖酵解酶，这表明代谢物通量而不是该途径的从头合成是在宿主细胞感染后触发的。在衣原体的细胞内形式网状体中也检测到细胞壁生物合成的中心酶，这表明肽聚糖在宿主细胞内生长期间产生。其他确定的蛋白质组包括17种在疫苗研究中具有潜在意义的外膜相关蛋白质和67种以前被注释为假设或保守假设的蛋白质。总的来说，C.沙眼衣原体的蛋白质组已得到实验验证，代表了迄今为止对任何衣原体蛋白质组的最广泛调查。
Chlamydiae are widespread bacterial pathogens responsible for a broad range of diseases, including sexually transmitted infections, pneumonia and trachoma. To validate the existence of hitherto hypothetical proteins predicted from recent chlamydial genome sequencing projects and to examine the patterns of expression of key components at the protein level, we have surveyed the expressed proteome of Chlamydia trachomatis strain L2. A combination of two-dimensional gel analysis, multi-dimensional protein identification (MudPIT) and nanocapillary liquid chromatography-tandem mass spectrometry allowed a total of 328 chlamydial proteins to be unambiguously assigned. Proteins identified as being expressed in the metabolically inert form, elementary body, of Chlamydia include the entire set of predicted glycolytic enzymes, indicating that metabolite flux rather than de novo synthesis of this pathway is triggered upon infection of host cells. An enzyme central to cell wall biosynthesis was also detected in the intracellular form, reticulate body, of Chlamydia, suggesting that the peptidoglycan is produced during growth within host cells. Other sets of proteins identified include 17 outer membrane-associated proteins of potential significance in vaccine studies and 67 proteins previously annotated as hypothetical or conserved hypothetical. Taken together, > 35% of the predicted proteome for C. trachomatis has been experimentally verified, representing the most extensive survey of any chlamydial proteome to date.