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首页> 外文期刊>Journal of Medical Microbiology: An Official Journal of the Pathological Society of Great Britain and Ireland >Reference map and comparative proteomic analysis of Neisseria gonorrhoeae displaying high resistance against spectinomycin
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Reference map and comparative proteomic analysis of Neisseria gonorrhoeae displaying high resistance against spectinomycin

机译：淋球菌对壮观霉素具有高抗性的参考图谱和比较蛋白质组学分析

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摘要

A proteome reference map of Neisseria gonorrhoeae was successfully established using two-dimensional gel electrophoresis in conjunction with matrix-assisted laser desorption ionization–time of flight mass spectrometry. This map was further applied to compare protein expression profiles of high-level spectinomycin-resistant (clinical isolate) and -susceptible (reference strain) N. gonorrhoeae following treatment with subminimal inhibitory concentrations (subMICs) of spectinomycin. Approximately 200 protein spots were visualized by Coomassie brilliant blue G-250 staining and 66 spots representing 58 unique proteins were subsequently identified. Most of the identified proteins were analysed as cytoplasmic proteins and belonged to the class of energy metabolism. Comparative proteomic analysis of whole protein expression of susceptible and resistant gonococci showed up to 96 % similarity while eight proteins were found to be differentially expressed in the resistant strain. In the presence of subMICs of spectinomycin, it was found that 50S ribosomal protein L7/L12, an essential component for ribosomal translocation, was upregulated in both strains, ranging from 1.5- to 3.5-fold, suggesting compensatory mechanisms of N. gonorrhoeae in response to antibiotic that inhibits protein synthesis. Moreover, the differential expression of proteins involved in energy metabolism, amino acid biosynthesis, and the cell envelope was noticeably detected, indicating significant cellular responses and adaptation against antibiotic stress. Such knowledge provides valuable data, not only fundamental proteomic data, but also knowledge of the mode of action of antibiotic and secondary target proteins implicated in adaptation and compensatory mechanisms.
机译：使用二维凝胶电泳结合基质辅助激光解吸电离-飞行时间质谱法成功建立了淋病奈瑟菌的蛋白质组参考图。将该图谱进一步用于比较经最小抑制浓度（subMICs）的壮观霉素治疗后对高壮观霉素耐药性（临床分离株）和敏感（参考菌株）淋病奈瑟氏球菌的蛋白质表达谱。通过考马斯亮蓝G-250染色观察到大约200个蛋白斑点，随后鉴定出代表58种独特蛋白的66个斑点。大部分鉴定出的蛋白质被分析为细胞质蛋白质，属于能量代谢类别。对易感和抗性淋球菌全蛋白表达的比较蛋白质组学分析显示出高达96％的相似性，而在抗性菌株中发现有8种蛋白差异表达。在壮观霉素的subMICs存在下，发现50S核糖体蛋白L7 / L12是核糖体易位的必需组分，在两种菌株中均被上调，范围是1.5-3.5倍，这表明淋病奈瑟氏球菌在应答中的补偿机制。抑制蛋白质合成的抗生素。此外，显着检测到参与能量代谢，氨基酸生物合成和细胞包膜的蛋白质的差异表达，这表明明显的细胞反应和对抗生素胁迫的适应性。这些知识不仅提供了基本的蛋白质组学数据，而且还提供了与适应和补偿机制有关的抗生素和次级靶蛋白的作用方式的知识。

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