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鱼皮黏液对鲑鱼弧菌可溶性蛋白质组的影响:二维凝胶电泳和串联质谱分析

Effect of fish skin mucus on the soluble proteome of Vibrio salmonicida analysed by 2-D gel electrophoresis and tandem mass spectrometry.

作者信息

Raeder Inger Lin Uttakleiv, Paulsen Steinar M, Smalås Arne O, Willassen Nils Peder

机构信息

Department of Chemistry, Faculty of Science, University of Tromsø, N-9037 Tromsø, Norway.

出版信息

Microb Pathog. 2007 Jan;42(1):36-45. doi: 10.1016/j.micpath.2006.10.003. Epub 2006 Dec 4.

Abstract

Vibrio salmonicida is the causative agent of cold-water vibriosis in farmed marine fish species. Adherence of pathogenic bacteria to mucosal surfaces is considered to be the first steps in the infective processes, and proteins involved are regarded as virulence factors. The global protein expression profile of V. salmonicida, grown with and without the presence of fish skin mucus in the synthetic media, was compared. Increased levels of proteins involved in motility, oxidative stress responses, and general stress responses were demonstrated as an effect of growth in the presence of mucus compared to non-mucus containing media. Enhanced levels of the flagellar proteins FlaC, FlaD and FlaE indicate increased motility capacity, while enhanced levels of the heat shock protein DnaK and the chaperonin GroEL indicate a general stress response. In addition, we observed that peroxidases, TPx.Grx and AhpC, involved in the oxidative stress responses, were induced by mucus proteins. The addition of mucus to the culture medium did not significantly alter the growth rate of V. salmonicida. An analysis of mucus proteins suggests that the mucus layer harbours a protein species that potentially possesses catalytic activity against DNA, and a protein with iron chelating activity. This study represents the first V. salmonicida proteomic analysis, and provides specific insight into the proteins necessary for the bacteria to challenge the skin mucus barrier of the fish.

摘要

杀鲑气单胞菌是养殖海水鱼类冷水弧菌病的病原体。病原菌粘附于粘膜表面被认为是感染过程的第一步,相关蛋白被视为毒力因子。比较了在合成培养基中有无鱼皮粘液存在的情况下杀鲑气单胞菌的全球蛋白质表达谱。与不含粘液的培养基相比,在有粘液存在的情况下生长,参与运动、氧化应激反应和一般应激反应的蛋白质水平升高。鞭毛蛋白FlaC、FlaD和FlaE水平的提高表明运动能力增强,而热休克蛋白DnaK和伴侣蛋白GroEL水平的提高表明存在一般应激反应。此外,我们观察到参与氧化应激反应的过氧化物酶TPx.Grx和AhpC被粘液蛋白诱导。向培养基中添加粘液并没有显著改变杀鲑气单胞菌的生长速率。对粘液蛋白的分析表明,粘液层含有一种可能具有DNA催化活性的蛋白质和一种具有铁螯合活性的蛋白质。本研究是首次对杀鲑气单胞菌进行蛋白质组学分析,为细菌突破鱼类皮肤粘液屏障所需的蛋白质提供了具体见解。

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