Department of Veterinary Biosciences, University of Helsinki, Helsinki, Finland.
Mol Cell Proteomics. 2011 Feb;10(2):M110.002741. doi: 10.1074/mcp.M110.002741. Epub 2010 Nov 15.
Lactobacillus rhamnosus GG (GG) is a widely used and intensively studied probiotic bacterium. Although the health benefits of strain GG are well documented, the systematic exploration of mechanisms by which this strain exerts probiotic effects in the host has only recently been initiated. The ability to survive the harsh conditions of the gastrointestinal tract, including gastric juice containing bile salts, is one of the vital characteristics that enables a probiotic bacterium to transiently colonize the host. Here we used gene expression profiling at the transcriptome and proteome levels to investigate the cellular response of strain GG toward bile under defined bioreactor conditions. The analyses revealed that in response to growth of strain GG in the presence of 0.2% ox gall the transcript levels of 316 genes changed significantly (p < 0.01, t test), and 42 proteins, including both intracellular and surface-exposed proteins (i.e. surfome), were differentially abundant (p < 0.01, t test in total proteome analysis; p < 0.05, t test in surfome analysis). Protein abundance changes correlated with transcriptome level changes for 14 of these proteins. The identified proteins suggest diverse and specific changes in general stress responses as well as in cell envelope-related functions, including in pathways affecting fatty acid composition, cell surface charge, and thickness of the exopolysaccharide layer. These changes are likely to strengthen the cell envelope against bile-induced stress and signal the GG cells of gut entrance. Notably, the surfome analyses demonstrated significant reduction in the abundance of a protein catalyzing the synthesis of exopolysaccharides, whereas a protein dedicated for active removal of bile compounds from the cells was up-regulated. These findings suggest a role for these proteins in facilitating the well founded interaction of strain GG with the host mucus in the presence of sublethal doses of bile. The significance of these findings in terms of the functionality of a probiotic bacterium is discussed.
鼠李糖乳杆菌 GG(GG)是一种广泛使用且经过深入研究的益生菌。尽管 GG 菌株的健康益处已有充分记录,但最近才开始系统探索该菌株在宿主中发挥益生菌作用的机制。能够在胃肠道的恶劣条件下生存,包括含有胆汁盐的胃液,是益生菌细菌能够短暂定植宿主的重要特征之一。在这里,我们使用转录组和蛋白质组水平的基因表达谱分析来研究 GG 菌株在特定生物反应器条件下对胆汁的细胞反应。分析表明,在含有 0.2%牛胆盐的条件下生长时,GG 菌株的 316 个基因的转录水平显著变化(p<0.01,t 检验),并且 42 种蛋白质,包括细胞内和表面暴露的蛋白质(即表面蛋白质组),丰度差异显著(p<0.01,总蛋白质组分析中的 t 检验;表面蛋白质组分析中的 p<0.05,t 检验)。这些蛋白质的丰度变化与其中 14 种蛋白质的转录水平变化相关。鉴定出的蛋白质表明,一般应激反应以及细胞包膜相关功能发生了多样化和特异性变化,包括影响脂肪酸组成、细胞表面电荷和胞外多糖层厚度的途径。这些变化可能会增强细胞膜对胆汁诱导的应激的抵抗力,并向 GG 细胞发出肠道入口的信号。值得注意的是,表面蛋白质组分析表明,参与合成胞外多糖的蛋白质丰度显著降低,而专门用于从细胞中主动去除胆汁化合物的蛋白质则上调。这些发现表明这些蛋白质在存在亚致死剂量胆汁的情况下促进 GG 菌株与宿主粘液的良好相互作用中发挥作用。讨论了这些发现对益生菌功能的意义。