Dynamic protein phosphorylation during the growth of Xanthomonas campestris pv. campestris B100 revealed by a gel-based proteomics approach.
Dynamic protein phosphorylation during the growth of Xanthomonas campestris pv. campestris B100 revealed by a gel-based proteomics approach.
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DOI：
10.1016/j.jbiotec.2013.06.009
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发表时间：
2013-08
影响因子：
4.1
通讯作者：
Y. Musa;Katrin Bäsell;Sarah Schatschneider;Frank-Jörg Vorhölter;D. Becher;K. Niehaus
中科院分区：
文献类型：
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作者：
Y. Musa;Katrin Bäsell;Sarah Schatschneider;Frank-Jörg Vorhölter;D. Becher;K. Niehaus
野油菜黄单胞菌野油菜致病变种（Xanthomonascampestris pv.campestris，Xcc）是一种能合成大量黄原胶的植物病原菌，其模式植物拟南芥（Arabidopsis thaliana）就是其中之一。黄原胶作为增稠剂通过Xcc的发酵在工业规模上生产。在基于二维凝胶电泳的方法中，来自不同生长阶段的蛋白质样品被表征以初始化黄单胞菌磷酸化蛋白质组的分析。用Pro-Q Diamond磷蛋白染色剂对2D凝胶进行染色以鉴定pueridine磷酸化蛋白。从凝胶上切下嘌呤磷酸化蛋白质的斑点，并通过质谱法进行分析。三种蛋白质被证实是磷酸化的，磷酸葡萄糖变位酶/磷酸甘露糖变位酶XanA，这是重要的黄原胶和脂多糖的生物合成，磷酸烯醇式丙酮酸合酶PspA，这是参与黄单胞菌，和抗σ因子拮抗剂RsbR是迄今为止未表征的黄单胞菌。收集样品的生长阶段对Xcc中的蛋白磷酸化有影响，特别是在RsbR的情况下，其在从指数生长后期到稳定期的过渡期间被磷酸化。
Xanthomonas campestris pv.campestris(Xcc) synthesizes huge amounts of the exopolysaccharide xanthan and is a plant pathogen affecting Brassicaceae, among them the model plant Arabidopsis thaliana. Xanthan is produced as a thickening agent at industrial scale by fermentation ofXcc. In an approach based on 2D gel electrophoresis, protein samples from different growth phases were characterized to initialize analysis of theXanthomonasphosphoproteome. The 2D gels were stained with Pro-Q Diamond phosphoprotein stain to identify putatively phosphorylated proteins. Spots of putatively phosphorylated proteins were excised from the gel and analyzed by mass spectrometry. Three proteins were confirmed to be phosphorylated, the phosphoglucomutase/phosphomannomutase XanA that is important for xanthan and lipopolysaccharide biosynthesis, the phosphoenolpyruvate synthase PspA that is involved in gluconeogenesis, and an anti-sigma factor antagonist RsbR that was so far uncharacterized in xanthomonads. The growth phase in which the samples were collected had an influence on protein phosphorylation inXcc, particular distinct in case of RsbR, which was phosphorylated during the transition from the late exponential growth phase to the stationary phase.