Analysis of the Pasteurella multocida outer membrane sub-proteome and its response to the in vivo environment of the natural host

Boyce, J. D., Cullen, P. A., Nguyen, V., Wilkie, I. and Adler, B. (2006) Analysis of the Pasteurella multocida outer membrane sub-proteome and its response to the in vivo environment of the natural host. Proteomics, 6 3: 870-880.


Author Boyce, J. D.
Cullen, P. A.
Nguyen, V.
Wilkie, I.
Adler, B.
Title Analysis of the Pasteurella multocida outer membrane sub-proteome and its response to the in vivo environment of the natural host
Journal name Proteomics  (ERA 2012 Listed)    (ERA 2010 Rank A)   Check publisher's open access policy
Publication date 2006
Sub-type Article
DOI 10.1002/pmic.200401342
Volume number 6
Issue number 3
ISSN 1615-9853
Start page 870
End page 880
Total pages 11
Place of publication Weinheim
Publisher Wiley-V C H Verlag Gmbh
Collection year 2006
Language eng
Subject C1
Abstract This study describes the identification of outer membrane proteins (OMPs) of the bacterial pathogen Pasteurella multocida and an analysis of how the expression of these proteins changes during infection of the natural host. We analysed the sarcosine-insoluble membrane fractions, which are highly enriched for OMPs, from bacteria grown under a range of conditions. Initially, the OMP-containing fractions were resolved by 2-DE and the proteins identified by MALDI-TOF MS. In addition, the OMP-containing fractions were separated by 1-D SDS-PAGE and protein identifications were made using nano LC MS/MS. Using these two methods a total of 35 proteins was identified from samples obtained from organisms grown in rich culture medium. Six of the proteins were identified only by 2-DE MALDI-TOF MS, whilst 17 proteins were identified only by 1-D LC MS/MS. We then analysed the OMPs from P. multocida which had been isolated from the bloodstream of infected chickens (a natural host) or grown in iron-depleted medium. Three proteins were found to be significantly up-regulated during growth in vivo and one of these (Pm0803) was also up-regulated during growth in iron-depleted medium. After bioinformatic analysis of the protein matches, it was predicted that over one third of the combined OMPs predicted by the bioinformatics sub-cellular localisation tools PSORTB and Proteome Analyst, had been identified during this study. This is the first comprehensive proteomic analysis of the P. multocida outer membrane and the first proteomic analysis of how a bacterial pathogen modifies its outer membrane proteome during infection.
Keyword 2-de
In Vivo Proteomics
Maldi Tof Ms
Nanolc-ms/ms
Outer Membrane
Biochemical Research Methods
Biochemistry & Molecular Biology
Elongation-factor Tu
Gram-negative Bacteria
Genomic-scale Analysis
Escherichia-coli
Fowl Cholera
Cross-protection
Gene-expression
Subcellular-localization
Haemophilus-influenzae
Proteins
Q-Index Code C1

 
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