In-depth analysis of exoproteomes from marine bacteria by shotgun liquid chromatography-tandem mass spectrometry: the Ruegeria pomeroyi DSS-3 case-study.
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.
Christie-Oleza
Joseph Alexander
JA
Laboratoire de Biochimie des Systèmes Perturbés, CEA Marcoule, DSV, iBEB, SBTN, LBSP, F-30207 BAGNOLS-SUR-CEZE, France.
Armengaud
Jean
J
D018528:ATP-Binding Cassette Transporters / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid; D009792:Oceans and Seas; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D021382:Protein Sorting Signals; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D041902:Rhodobacteraceae / Q000737:chemistry* / Q000378:metabolism; D012623:Seawater; D053719:Tandem Mass Spectrometry
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D020543:Proteome