title: Proteome Profiles of Outer Membrane Vesicles and Extracellular Matrix of Pseudomonas aeruginosa Biofilms.
abstract: In the present work, two different proteomic platforms, gel-based and gel-free, were used to map the matrix and outer membrane vesicle exoproteomes of Pseudomonas aeruginosa PAO1 biofilms. These two proteomic strategies allowed us a confident identification of 207 and 327 proteins from enriched outer membrane vesicles and whole matrix isolated from biofilms. Because of the physicochemical characteristics of these subproteomes, the two strategies showed complementarity, and thus, the most comprehensive analysis of P. aeruginosa exoproteome to date was achieved. Under our conditions, outer membrane vesicles contribute approximately 20% of the whole matrix proteome, demonstrating that membrane vesicles are an important component of the matrix. The proteomic profiles were analyzed in terms of their biological context, namely, a biofilm. Accordingly relevant metabolic processes involved in cellular adaptation to the biofilm lifestyle as well as those related to P. aeruginosa virulence capabilities were a key feature of the analyses. The diversity of the matrix proteome corroborates the idea of high heterogeneity within the biofilm; cells can display different levels of metabolism and can adapt to local microenvironments making this proteomic analysis challenging. In addition to analyzing our own primary data, we extend the analysis to published data by other groups in order to deepen our understanding of the complexity inherent within biofilm populations.
authors: Michael Barber Centre for Mass Spectrometry, Manchester Institute for Biotechnology, University of Manchester , Princess Road, Manchester, M1 7DN, U.K.
authors: Narciso
authors: N
authors: Couto
authors: Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph , Guelph, ON N1G 2W1, Canada.|Department of Physics, University of Guelph , Guelph, ON N1G 2W1, Canada.
authors: Sarah R
authors: SR
authors: Schooling
authors: Department of Physics, University of Guelph , Guelph, ON N1G 2W1, Canada.
authors: John R
authors: JR
authors: Dutcher
authors: Michael Barber Centre for Mass Spectrometry, Manchester Institute for Biotechnology, University of Manchester , Princess Road, Manchester, M1 7DN, U.K.|Manchester Pharmacy School, University of Manchester , Stopford Building, Oxford Road, Manchester, M13 9PT, U.K.
authors: Jill
authors: J
authors: Barber
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D018441:Biofilms / Q000254:growth & development*; D002462:Cell Membrane / Q000737:chemistry; D002853:Chromatography, Liquid; D005109:Extracellular Matrix / Q000737:chemistry*; D000067128:Extracellular Vesicles / Q000737:chemistry*; D005782:Gels; D015964:Gene Expression Regulation, Bacterial; D058977:Molecular Sequence Annotation; D020543:Proteome / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D011550:Pseudomonas aeruginosa / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D037521:Virulence Factors / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D005782:Gels; D020543:Proteome; D037521:Virulence Factors
keywords: Pseudomonas aeruginosa; biofilms; matrix; outer membrane vesicles; proteome