Analysis of the periplasmic proteome of Pseudomonas aeruginosa, a metabolically versatile opportunistic pathogen
Analysis of the periplasmic proteome of Pseudomonas aeruginosa, a metabolically versatile opportunistic pathogen
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DOI：
10.1002/pmic.200800618
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发表时间：
2009-04-01
期刊：
影响因子：
3.4
通讯作者：
Fimia, Gian Maria
中科院分区：
文献类型：
--
作者：
Imperi, Francesco;Ciccosanti, Fabiola;Fimia, Gian Maria
革兰氏阴性菌铜绿假单胞菌是住院、烧伤、免疫功能低下和囊性纤维化患者感染的主要原因。铜绿假单胞菌致病所必需的许多过程，例如，营养吸收、抗生素抗性和毒力发生在细胞被膜中，并依赖于驻留在周质空间中的组分。最近的高通量研究集中在铜绿假单胞菌膜隔室。然而，其周质的组成和动力学仍然在很大程度上没有特征。在这里，我们报告的周质蛋白质组的2-DE和MALDI-TOF/TOF分析的野生型铜绿假单胞菌菌株PAO 1的详细描述。在蛋白质组水平上比较了三种提取方法，以获得最可靠和最全面的周质蛋白质图谱。共鉴定了495个点，代表395种不同的蛋白质。大部分的高强度点对应于周质蛋白，而细胞质污染物主要检测微弱的斑点。大多数已鉴定的周质蛋白参与转运、细胞被膜完整性和蛋白质折叠控制。值得注意的是，超过30%仍然具有不可预测的功能。这项工作提供了铜绿假单胞菌周质的第一个概述，并为未来研究铜绿假单胞菌适应不同环境和/或抗生素治疗期间发生的周质蛋白质组变化提供了基础。
The Gram-negative bacterium Pseudomonas aeruginosa is a main cause of infection in hospitalized, burned, immunocompromised, and cystic fibrosis patients. Many processes essential for P. aeruginosa pathogenesis, e.g., nutrient uptake, antibiotic resistance, and virulence, take place in the cell envelope and depend on components residing in the periplasmic space. Recent high-throughput studies focused on P. aeruginosa membrane compartments. However, the composition and dynamics of its periplasm remain largely uncharacterized. Here, we report a detailed description of the periplasmic proteome of the wild-type P. aeruginosa strain PAO1 by 2-DE and MALDI-TOF/TOF analysis. Three extraction methods were compared at proteome level in order to achieve the most reliable and comprehensive periplasmic protein map. A total of 495 spots representing 395 different proteins were identified. Most of the high intensity spots corresponded to periplasmic proteins, while cytoplasmic contaminants were mainly detected among faint spots. The majority, of the identified periplasmic proteins is involved in transport, cell envelope integrity, and protein folding control. Notably, more than 30% still has an unpredicted function. This work provides the first overview of the P. aeruginosa periplasm and offers the basis for future studies on periplasmic proteome changes occurring during P. aeruginosa adaptation to different environments and/or antibiotic treatments.