Assessing the exoproteome of marine bacteria, lesson from a RTX-toxin abundantly secreted by Phaeobacter strain DSM 17395

PLoS One. 2014 Feb 24;9(2):e89691. doi: 10.1371/journal.pone.0089691. eCollection 2014.

Abstract

Bacteria from the Roseobacter clade are abundant in surface marine ecosystems as over 10% of bacterial cells in the open ocean and 20% in coastal waters belong to this group. In order to document how these marine bacteria interact with their environment, we analyzed the exoproteome of Phaeobacter strain DSM 17395. We grew the strain in marine medium, collected the exoproteome and catalogued its content with high-throughput nanoLC-MS/MS shotgun proteomics. The major component represented 60% of the total protein content but was refractory to either classical proteomic identification or proteogenomics. We de novo sequenced this abundant protein with high-resolution tandem mass spectra which turned out being the 53 kDa RTX-toxin ZP_02147451. It comprised a peptidase M10 serralysin domain. We explained its recalcitrance to trypsin proteolysis and proteomic identification by its unusual low number of basic residues. We found this is a conserved trait in RTX-toxins from Roseobacter strains which probably explains their persistence in the harsh conditions around bacteria. Comprehensive analysis of exoproteomes from environmental bacteria should take into account this proteolytic recalcitrance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Electrophoresis, Polyacrylamide Gel
  • Marine Toxins / chemistry
  • Marine Toxins / isolation & purification
  • Marine Toxins / metabolism
  • Molecular Sequence Data
  • Proteomics / methods
  • Rhodobacteraceae / chemistry
  • Rhodobacteraceae / genetics*
  • Roseobacter / genetics
  • Seawater / microbiology
  • Tandem Mass Spectrometry
  • Trypsin / metabolism

Substances

  • Marine Toxins
  • Trypsin

Grants and funding

JA Christie-Oleza was supported by a fellowship from the Fundación Ramón Areces. The authors thank the Commissariat à l’Energie Atomique et aux Energies Alternatives for financial support. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.