Heim S, Ferrer M, Heuer H, Regenhardt D, Nimtz M, Timmis K N
Department of Environmental Microbiology, GBF--German Research Centre for Biotechnology, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Environ Microbiol. 2003 Dec;5(12):1257-69. doi: 10.1111/j.1462-2920.2003.00465.x.
The genome sequence of Pseudomonas putida strain KT2440, a nutritionally versatile, saprophytic and plant root-colonizing Gram-negative soil bacterium, was recently determined by K. E. Nelson et al. (2002, Environ Microbiol 4: 799-808). Here, we present a two-dimensional gel protein reference map of KT2440 cells grown in mineral salts medium with glucose as carbon source. Proteins were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) analysis, in conjunction with an in-house database developed from the genome sequence of KT2440, and approximately 200 two-dimensional gel spots were assigned. The map was used to assess the genomic response of KT2440 to iron limitation stress and to compare this response with that of the closely related facultative human pathogen Pseudomonas aeruginosa strain PAO1. The synthesis of about 25 proteins was affected in both strains, including four prominent upregulated ferric uptake regulator (Fur) protein-dependent proteins, but there were also striking differences in their proteome responses, for example in the expression of superoxide dismutases (Sod), which may indicate important roles of iron-responsive functions in the adaptation of these two bacteria to different lifestyles. The Sod enzyme of KT2440 was shown to be a novel heterodimer of the SodA and SodB polypeptides.
恶臭假单胞菌KT2440是一种营养需求多样、腐生且能定殖于植物根部的革兰氏阴性土壤细菌,其基因组序列最近由K. E. 尼尔森等人测定(2002年,《环境微生物学》4: 799 - 808)。在此,我们展示了以葡萄糖作为碳源在无机盐培养基中生长的KT2440细胞的二维凝胶蛋白质参考图谱。蛋白质通过基质辅助激光解吸电离飞行时间(MALDI - TOF)分析进行鉴定,并结合从KT2440基因组序列开发的内部数据库,大约有200个二维凝胶斑点被成功鉴定。该图谱用于评估KT2440对铁限制应激的基因组反应,并将此反应与密切相关的兼性人类病原体铜绿假单胞菌PAO1的反应进行比较。两种菌株中约25种蛋白质的合成受到影响,包括四种显著上调的铁摄取调节因子(Fur)蛋白依赖性蛋白质,但它们的蛋白质组反应也存在显著差异,例如超氧化物歧化酶(Sod)的表达,这可能表明铁响应功能在这两种细菌适应不同生活方式中发挥着重要作用。KT2440的Sod酶被证明是SodA和SodB多肽的新型异源二聚体。