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两株长双歧杆菌的无标记定量蛋白质组学

Label-free quantitative proteomics of two Bifidobacterium longum strains.

作者信息

Guillaume Elisabeth, Berger Bernard, Affolter Michael, Kussmann Martin

机构信息

Functional Genomics Group, Department of BioAnalytical Sciences, Nestlé Research Centre, Nestec Ltd., Vers-chez-les-Blanc, P.O. Box 44, CH-1000 Lausanne 26, Switzerland.

出版信息

J Proteomics. 2009 Jul 21;72(5):771-84. doi: 10.1016/j.jprot.2009.03.004. Epub 2009 Mar 26.

Abstract

Bifidobacteria are gram-positive anaerobes with a high Guanine/Cytosine genome content. Specific strains are used as probiotics because of their health benefits. Probiotics are live microorganisms with a positive influence on their host's health. They are used as nutritional supplements and their resistance to conditions of food manufacturing and of the gastro-intestinal tract is studied. We report on differential proteomics of two Bifidobacterium longum strains that differ in their heat shock resistance. This was achieved by a comparative qualitative survey of their proteomes and relative LC-MS/MS-based label-free protein quantification. Deploying a nano LC-ESI ion-trap mass spectrometer, 165 proteins expressed by the two probiotic strains were identified. Around 50% of these were common to both strains with the remaining half identified in either one of the strains. Using a label-free technology based on the 3D overlay of retention times, mass-over-charge ratios and ion intensities between LC-MS runs, we found quantitative differences in the relative abundance of 19 of the proteins common to both strains. The differentially expressed proteins were classified into categories involving glucose metabolism, protein synthesis and heat shock proteins. Six of these 19 proteins have been previously reported as being associated to heat stress response.

摘要

双歧杆菌是革兰氏阳性厌氧菌,其基因组中鸟嘌呤/胞嘧啶含量较高。特定菌株因其对健康有益而被用作益生菌。益生菌是对宿主健康有积极影响的活微生物。它们被用作营养补充剂,并对其在食品制造和胃肠道条件下的抗性进行研究。我们报告了两种抗热休克能力不同的长双歧杆菌菌株的差异蛋白质组学。这是通过对它们的蛋白质组进行比较定性调查以及基于液相色谱-串联质谱的无标记蛋白质相对定量来实现的。使用纳升液相色谱-电喷雾离子阱质谱仪,鉴定出这两种益生菌菌株表达的165种蛋白质。其中约50%是两种菌株共有的,其余一半在其中一种菌株中鉴定出来。使用基于液相色谱-质谱运行之间保留时间、质荷比和离子强度的三维叠加的无标记技术,我们发现两种菌株共有的19种蛋白质的相对丰度存在定量差异。差异表达的蛋白质被分类为涉及葡萄糖代谢、蛋白质合成和热休克蛋白的类别。这19种蛋白质中有6种先前已被报道与热应激反应有关。

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