Sarma Annamraju D, Emerich David W
Department of Biochemistry, University of Missouri, Columbia, MO 65211, USA.
Proteomics. 2005 Nov;5(16):4170-84. doi: 10.1002/pmic.200401296.
As a prelude to using functional proteomics towards understanding the process of symbiotic nitrogen fixation between the legume soybean and the soil bacteria Bradyrhizobium japonicum, we examined the total protein expression pattern of the nodule bacteria, often referred to as bacteroids. A partial proteome map was constructed by separating the total bacteroid proteins using high-resolution 2-DE. Of the several hundred protein spots analyzed using PMF, 180 spots were tentatively identified by searching the available database for B. japonicum, (http://www.kazusa.or.jp/index.html). The data showed that the bacteroid expressed a dominant and elaborate protein network for nitrogen and carbon metabolism, which is closely dependent on the plant supplied metabolites, and seems aptly supported by a selective group of bacteroid transporter proteins. However, they seem to lack a defined fatty acid and nucleic acid metabolism. Interestingly, the proteins related to protein synthesis, scaffolding and degradation were among the most predominant spots of the bacteroid proteome. In addition, several proteins, which showed fairly good expression, were identified to be involved with cellular detoxification, stress regulation and signaling communication components. This preliminary proteomic data matches very well with several biochemical and genetic reports, and clearly shows the inter-connection between several metabolic pathways that meet the needs of the bacteroid. It is expected that in the future this will allow us to develop testable hypotheses about the roles of several of these proteins in context to the metabolic pathway connections and metabolite fluxes.
作为运用功能蛋白质组学来理解豆科植物大豆与土壤细菌日本慢生根瘤菌之间共生固氮过程的前奏,我们检测了根瘤菌(通常称为类菌体)的总蛋白表达模式。通过使用高分辨率二维电泳分离类菌体总蛋白构建了部分蛋白质组图谱。在使用基质辅助激光解吸电离飞行时间质谱分析的数百个蛋白点中,通过搜索日本慢生根瘤菌的可用数据库(http://www.kazusa.or.jp/index.html)初步鉴定出180个点。数据表明,类菌体表达了一个用于氮和碳代谢的主导且精细的蛋白质网络,该网络紧密依赖于植物提供的代谢物,并且似乎由一组选择性的类菌体转运蛋白提供了恰当的支持。然而,它们似乎缺乏明确的脂肪酸和核酸代谢。有趣的是,与蛋白质合成、支架构建和降解相关的蛋白质是类菌体蛋白质组中最主要的斑点之一。此外,鉴定出几种表达相当良好的蛋白质参与细胞解毒、应激调节和信号通讯成分。这些初步的蛋白质组学数据与几份生化和遗传学报告非常吻合,并且清楚地显示了满足类菌体需求的几种代谢途径之间的相互联系。预计未来这将使我们能够针对这些蛋白质中的几种在代谢途径连接和代谢物通量方面的作用提出可检验的假设。