PDF(Pubmed)
Abstract：
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.
摘要：
微生物向它们的细胞外环境分泌许多它们生存所需的化合物。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非常感兴趣。分泌的蛋白质和易位系统本身的成分可以通过最新的蛋白质组学工具进行深入的检查。虽然病原体的分泌组是有据可查的，那些非病原体仍在很大程度上有待建立。这里,我们提供了在标准实验室条件下生长的海洋细菌RuegeriapomeroyiDSS-3的外蛋白质组分析。我们使用了shot弹枪方法，包括外蛋白质组的胰蛋白酶消化，并通过液相色谱与串联质谱联用鉴定所得肽。揭示了具有与RTX毒素中观察到的那些同源的结构域的三种不同蛋白质,并将其半定量为分泌最丰富的蛋白质。其中一种蛋白质显然从目录中脱颖而出，代表超过一半的总的外蛋白质组。我们还列出了许多与ABC和TRAP转运蛋白相关的可溶性蛋白质，这些蛋白质暗示了营养素的摄取。RuegeriapomeroyiDSS-3案例研究说明了shot弹枪纳米LC-MS/MS策略破译海洋细菌外蛋白质组并为环境蛋白质组学做出贡献的能力。
