title: Pseudomonas aeruginosa Cell Membrane Protein Expression from Phenotypically Diverse Cystic Fibrosis Isolates Demonstrates Host-Specific Adaptations.
abstract: Pseudomonas aeruginosa is a Gram-negative, nosocomial, highly adaptable opportunistic pathogen especially prevalent in immuno-compromised cystic fibrosis (CF) patients. The bacterial cell surface proteins are important contributors to virulence, yet the membrane subproteomes of phenotypically diverse P. aeruginosa strains are poorly characterized. We carried out mass spectrometry (MS)-based proteome analysis of the membrane proteins of three novel P. aeruginosa strains isolated from the sputum of CF patients and compared protein expression to the widely used laboratory strain, PAO1. Microbes were grown in planktonic growth condition using minimal M9 media, and a defined synthetic lung nutrient mimicking medium (SCFM) limited passaging. Two-dimensional LC-MS/MS using iTRAQ labeling enabled quantitative comparisons among 3171 and 2442 proteins from the minimal M9 medium and in the SCFM, respectively. The CF isolates showed marked differences in membrane protein expression in comparison with PAO1 including up-regulation of drug resistance proteins (MexY, MexB, MexC) and down-regulation of chemotaxis and aerotaxis proteins (PA1561, PctA, PctB) and motility and adhesion proteins (FliK, FlgE, FliD, PilJ). Phenotypic analysis using adhesion, motility, and drug susceptibility assays confirmed the proteomics findings. These results provide evidence of host-specific microevolution of P. aeruginosa in the CF lung and shed light on the adaptation strategies used by CF pathogens.
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Karthik Shantharam
authors.initials: KS
authors.lastname: Kamath
authors.affiliation: Australian Proteome Analysis Facility, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Dana
authors.initials: D
authors.lastname: Pascovici
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Anahit
authors.initials: A
authors.lastname: Penesyan
authors.affiliation: Australian Proteome Analysis Facility, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Apurv
authors.initials: A
authors.lastname: Goel
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Vignesh
authors.initials: V
authors.lastname: Venkatakrishnan
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Ian T
authors.initials: IT
authors.lastname: Paulsen
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Nicolle H
authors.initials: NH
authors.lastname: Packer
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University , Sydney, New South Wales 2109, Australia.|Australian Proteome Analysis Facility, Macquarie University , Sydney, New South Wales 2109, Australia.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Molloy
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics*; D001426:Bacterial Proteins / Q000032:analysis; D002853:Chromatography, Liquid; D003550:Cystic Fibrosis / Q000382:microbiology*; D015964:Gene Expression Regulation, Bacterial; D054884:Host-Pathogen Interactions; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis*; D010641:Phenotype; D040901:Proteomics / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000737:chemistry*; D013183:Sputum / Q000382:microbiology; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins
keywords: Pseudomonas aeruginosa; bacterial evolution and adaptation; mass spectrometry; membrane proteome; proteomics; virulence