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首页> 外文期刊>Marine Drugs >In-Depth Analysis of Exoproteomes from Marine Bacteria by Shotgun Liquid Chromatography-Tandem Mass Spectrometry: the Ruegeria pomeroyi DSS-3 Case-Study
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In-Depth Analysis of Exoproteomes from Marine Bacteria by Shotgun Liquid Chromatography-Tandem Mass Spectrometry: the Ruegeria pomeroyi DSS-3 Case-Study

机译：散弹枪液相色谱-串联质谱对海洋细菌外蛋白质组进行深入分析：Ruegeria pomeroyi DSS-3案例研究

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Microorganisms secrete into their extracellular environment numerous compounds that are required for their survival. Many of these compounds could be of great interest for biotechnology applications and their genes used in synthetic biology design. The secreted proteins and the components of the translocation systems themselves can be scrutinized in-depth by the most recent proteomic tools. While the secretomes of pathogens are well-documented, those of non-pathogens remain largely to be established. Here, we present the analysis of the exoproteome from the marine bacterium Ruegeria pomeroyi DSS-3 grown in standard laboratory conditions. We used a shotgun approach consisting of trypsin digestion of the exoproteome, and identification of the resulting peptides by liquid chromatography coupled to tandem mass spectrometry. Three different proteins that have domains homologous to those observed in RTX toxins were uncovered and were semi-quantified as the most abundantly secreted proteins. One of these proteins clearly stands out from the catalogue, representing over half of the total exoproteome. We also listed many soluble proteins related to ABC and TRAP transporters implied in the uptake of nutrients. The Ruegeria pomeroyi DSS-3 case-study illustrates the power of the shotgun nano-LC-MS/MS strategy to decipher the exoproteome from marine bacteria and to contribute to environmental proteomics.
机译：微生物将多种生存所需的化合物分泌到其细胞外环境中。这些化合物中的许多对于生物技术应用及其在合成生物学设计中使用的基因可能都非常重要。分泌蛋白和转运系统本身的组成部分可以通过最新的蛋白质组学工具进行深入研究。尽管病原体的分泌组有充分的文献记载，但非病原体的分泌组仍在很大程度上建立。在这里，我们介绍了在标准实验室条件下生长的海洋细菌Ruegeria pomeroyi DSS-3的外蛋白质组的分析。我们使用a弹枪方法，包括胰蛋白酶消化外蛋白质组，以及通过液相色谱-串联质谱法鉴定所得肽。结构域与RTX毒素中观察到的结构域同源的三种不同蛋白质未被发现，并被半定量为分泌最丰富的蛋白质。这些蛋白质中的一种显然从目录中脱颖而出，占外蛋白质组总数的一半以上。我们还列出了许多与营养吸收有关的与ABC和TRAP转运蛋白有关的可溶性蛋白。 Ruegeria pomeroyi DSS-3案例研究说明了gun弹枪纳米LC-MS / MS策略从海洋细菌中破译蛋白质组并为环境蛋白质组学做出贡献的能力。

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