PSEAE oprF.2012

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This is supplementary website for the following paper:

Aimee K. Wessel, Jean Liew, Taejoon Kwon, Edward M. Marcotte, Marvin Whiteley, The role of Pseudomonas aeruginosa peptidoglycan-associated outer membrane proteins in vesicle biogenesis, Submitted.



Gram-negative bacteria produce outer membrane vesicles (OMVs) that package and deliver proteins, small molecules, and DNA to prokaryotic and eukaryotic cells. The molecular details of OMV biogenesis have not been fully elucidated, but peptidoglycan-associated outer membrane proteins that tether the outer membrane to the underlying peptidoglycan have been shown to be critical for OMV formation in multiple Enterobacteriaceae. In this study, we demonstrate that the peptidoglycan-associated outer membrane proteins OprF and OprI, but not OprL, contribute to OMV formation in the opportunistic pathogen Pseudomonas aeruginosa. Interestingly, OprF 9 does not appear to be important for tethering the outer membrane to peptidoglycan but instead impacts OMV formation through modulation of the levels of the Pseudomonas Quinolone Signal (PQS), a quorum signal previously shown by our laboratory to be critical for OMV formation. Thus the role of OprF in OMV biogenesis is distinct from other peptidoglycan-associated outer 13 membrane proteins including OprI.

Shotgun proteomics experiment design

  • Samples
  1. PA14OMV - outer membrane vesicles from P. aeruginosa PA14 wild-type
  2. PA14OMVoprF - outer membrane vesicles from P. aeruginosa PA14 oprF deletion mutant
  • Two injections per sample, with Thermo Orbitrap Velos. 28,000 ~ 36,000 ms2 spectra were collected per injection.

Shotgun proteomics data

Database search result