Outer membrane proteome of Actinobacillus pleuropneumoniae: LC-MS/MS analyses validate in silico predictions

The Gram‐negative bacterial pathogen Actinobacillus pleuropneumoniae causes porcine pneumonia, a highly infectious respiratory disease that contributes to major economic losses in the swine industry. Outer membrane (OM) proteins play key roles in infection and may be targets for drug and vaccine res...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2007-06, Vol.7 (11), p.1854-1865
Main Authors: Chung, Jacqueline W., Ng-Thow-Hing, Christopher, Budman, Lorne I., Gibbs, Bernard F., Nash, John H. E., Jacques, Mario, Coulton, James W.
Format: Article
Language:English
Subjects:
Actinobacillus pleuropneumoniae
Actinobacillus pleuropneumoniae - chemistry
Actinobacillus pleuropneumoniae - genetics
Actinobacillus pleuropneumoniae - metabolism
Analytical, structural and metabolic biochemistry
Bacterial Outer Membrane Proteins - analysis
Bacterial Outer Membrane Proteins - genetics
Bacterial Outer Membrane Proteins - metabolism
Biological and medical sciences
Chromatography, Liquid
Computational Biology
Electrophoresis, Gel, Two-Dimensional
Extraction protocols
Fundamental and applied biological sciences. Psychology
LC-MS/MS
Lipoproteins - analysis
Lipoproteins - genetics
Lipoproteins - metabolism
Mass Spectrometry
Miscellaneous
Outer membrane proteome
Proteins
Proteomics - methods
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Summary:The Gram‐negative bacterial pathogen Actinobacillus pleuropneumoniae causes porcine pneumonia, a highly infectious respiratory disease that contributes to major economic losses in the swine industry. Outer membrane (OM) proteins play key roles in infection and may be targets for drug and vaccine research. Exploiting the genome sequence of A. pleuropneumoniae serotype 5b, we scanned in silico for proteins predicted to be localized at the cell surface. Five genome scanning programs (Proteome Analyst, PSORT‐b, BOMP, Lipo, and LipoP) were run to construct a consensus prediction list of 93 OM proteins in A. pleuropneumoniae. An inventory of predicted OM proteins was complemented by proteomic analyses utilizing gel‐ and solution‐based methods, both coupled to LC‐MS/MS. Different protocols were explored to enrich for OM proteins; the most rewarding required sucrose gradient centrifugation followed by membrane washes with sodium bromide and sodium carbonate. This protocol facilitated our identification of 47 OM proteins that represent 50% of the predicted OM proteome, most of which have not been characterized. Our study establishes the first OM proteome of A. pleuropneumoniae.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200600979