Al Dahouk Sascha, Jubier-Maurin Véronique, Scholz Holger C, Tomaso Herbert, Karges Wolfram, Neubauer Heinrich, Köhler Stephan
Centre d'Etudes d'Agents Pathogènes et Biotechnologies pour la Santé (CPBS), Université Montpellier 1, Montpellier, France.
Proteomics. 2008 Sep;8(18):3862-70. doi: 10.1002/pmic.200800026.
A 2-D DIGE approach allowed the characterization of the intramacrophagic proteome of the intracellular pathogen Brucella suis at the late stage of in vitro infection by efficient discrimination between bacterial and host cell proteins. Using a subtraction model, a total of 168 proteins showing altered concentrations in comparison with extracellularly grown, stationary-phase bacteria were identified. The majority of the 44 proteins significantly regulated at this stage of infection were involved in bacterial metabolism and 40% were present in lowered concentrations, supporting the hypothesis of an adaptive response by quantitative reduction of processes participating in energy, protein, and nucleic acid metabolism. In the future, the 2-D DIGE-based approach will permit to decipher specifically and quantitatively the intracellular proteomes of various pathogens during adaptation to their specific host cell environments.
二维差异凝胶电泳(2-D DIGE)方法通过有效区分细菌和宿主细胞蛋白,实现了对细胞内病原体猪布鲁氏菌体外感染后期巨噬细胞内蛋白质组的表征。使用消减模型,共鉴定出168种与细胞外生长的稳定期细菌相比浓度发生变化的蛋白质。在感染的这个阶段,44种显著调节的蛋白质中,大多数参与细菌代谢,40%的蛋白质浓度降低,这支持了通过定量减少参与能量、蛋白质和核酸代谢的过程来进行适应性反应的假说。未来,基于二维差异凝胶电泳的方法将能够在各种病原体适应其特定宿主细胞环境的过程中,特异性地、定量地解析细胞内蛋白质组。