Pyndiah Slovénie, Lasserre Jean Paul, Ménard Armelle, Claverol Stéphane, Prouzet-Mauléon Valérie, Mégraud Francis, Zerbib Frank, Bonneu Marc
INSERM, U853, Bordeaux, F 33076 France.
Mol Cell Proteomics. 2007 Feb;6(2):193-206. doi: 10.1074/mcp.M600363-MCP200. Epub 2006 Nov 7.
The study of protein interactions constitutes an important domain to understand the physiology and pathogenesis of microorganisms. The two-dimensional blue native/SDS-PAGE was initially reported to analyze membrane protein complexes. In this study, both cytoplasmic and membrane complexes of a bacterium, the strain J99 of the gastric pathogen Helicobacter pylori, were analyzed by this method. It was possible to identify 34 different proteins grouped in 13 multiprotein complexes, 11 from the cytoplasm and two from the membrane, either previously reported partially or totally in the literature. Besides complexes involved in H. pylori physiology, this method allowed the description of interactions involving known pathogenic factors such as (i) urease with the heat shock protein GroEL or with the putative ketol-acid reductoisomerase IlvC and (ii) the cag pathogenicity island CagA protein with the DNA gyrase GyrA as well as insight on the partners of TsaA, a peroxide reductase/stress-dependent molecular chaperone. The two-dimensional blue native/SDS-PAGE combined with mass spectrometry is a potential tool to study the differences in complexes isolated in various situations and also to study the interactions between bacterial and eucaryotic cell proteins.
蛋白质相互作用的研究是理解微生物生理学和发病机制的一个重要领域。二维蓝色天然/SDS-PAGE最初被报道用于分析膜蛋白复合物。在本研究中，采用该方法对一种细菌——胃病原体幽门螺杆菌J99菌株的细胞质和膜复合物进行了分析。通过该方法可以鉴定出34种不同的蛋白质，它们被归为13个多蛋白复合物，其中11个来自细胞质，2个来自膜，这些复合物在文献中部分或全部已有报道。除了参与幽门螺杆菌生理学的复合物外，该方法还揭示了涉及已知致病因子的相互作用，如（i）脲酶与热休克蛋白GroEL或假定的酮醇酸还原异构酶IlvC的相互作用，以及（ii）cag致病岛CagA蛋白与DNA促旋酶GyrA的相互作用，同时还深入了解了过氧化物还原酶/应激依赖性分子伴侣TsaA的相互作用伙伴。二维蓝色天然/SDS-PAGE与质谱联用是研究不同情况下分离的复合物差异以及细菌和真核细胞蛋白之间相互作用的一种潜在工具。
