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Towards a Rigorous Network of Protein-Protein Interactions of the Model Sulfate Reducer Desulfovibrio vulgaris Hildenborough

机译:迈向模型硫酸盐还原剂Desulfovibrio vulgaris Hildenborough的蛋白质-蛋白质相互作用的严格网络

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摘要

Protein–protein interactions offer an insight into cellular processes beyond what may be obtained by the quantitative functional genomics tools of proteomics and transcriptomics. The aforementioned tools have been extensively applied to study Escherichia coli and other aerobes and more recently to study the stress response behavior of Desulfovibrio vulgaris Hildenborough, a model obligate anaerobe and sulfate reducer and the subject of this study. Here we carried out affinity purification followed by mass spectrometry to reconstruct an interaction network among 12 chromosomally encoded bait and 90 prey proteins based on 134 bait-prey interactions identified to be of high confidence. Protein-protein interaction data are often plagued by the lack of adequate controls and replication analyses necessary to assess confidence in the results, including identification of potential false positives. We addressed these issues through the use of biological replication, exponentially modified protein abundance indices, results from an experimental negative control, and a statistical test to assign confidence to each putative interacting pair applicable to small interaction data studies. We discuss the biological significance of metabolic features of D. vulgaris revealed by these protein-protein interaction data and the observed protein modifications. These include the distinct role of the putative carbon monoxide-induced hydrogenase, unique electron transfer routes associated with different oxidoreductases, and the possible role of methylation in regulating sulfate reduction.
机译:蛋白质间相互作用不仅可以通过蛋白质组学和转录组学的定量功能基因组学工具获得深入的了解,还可以深入了解细胞过程。前述工具已被广泛地用于研究大肠杆菌和其他需氧菌,并且最近被用于研究Desulfovibrio vulgaris Hildenborough的应激反应行为,该模型为专性厌氧菌和硫酸盐还原剂,是本研究的主题。在这里,我们进行了亲和纯化,然后进行了质谱分析,以基于134个诱饵与猎物之间的相互作用确定了12个染色体编码的诱饵与90个猎物蛋白之间的相互作用网络,从而确定了它们的高可信度。蛋白质-蛋白质相互作用数据常常因缺乏评估结果可信度所需的适当对照和复制分析而困扰,包括确定潜在的假阳性结果。我们通过使用生物复制,指数修饰的蛋白质丰度指数,实验阴性对照的结果以及统计检验为每个适用于小型相互作用数据研究的推定相互作用对分配置信度来解决这些问题。我们讨论了由这些蛋白质-蛋白质相互作用数据和观察到的蛋白质修饰所揭示的D. v。寻常代谢特征的生物学意义。这些包括推定的一氧化碳诱导的氢化酶的独特作用,与不同的氧化还原酶相关的独特的电子转移途径以及甲基化在调节硫酸盐还原中的可能作用。

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