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Journal of Biomedicine and Biotechnology
Volume 2011 (2011), Article ID 701928, 10 pages
http://dx.doi.org/10.1155/2011/701928
Research Article

Proteomics Analyses of the Opportunistic Pathogen Burkholderia vietnamiensis Using Protein Fractionations and Mass Spectrometry

1Department of Chemistry and Biochemistry, University of Arizona, 1306 E University Bolevard, Tucson, AZ 85721, USA
2Department of Soil Water and Environmental Science, University of Arizona, Tucson, AZ 85721, USA

Received 2 June 2011; Revised 27 August 2011; Accepted 27 August 2011

Academic Editor: Isabel Sá-Correia

Copyright © 2011 Samanthi Wickramasekara et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The main objectives of this work were to obtain a more extensive coverage of the Burkholderia vietnamiensis proteome than previously reported and to identify virulence factors using tandem mass spectrometry. The proteome of B. vietnamiensis was precipitated into four fractions to as extracellular, intracellular, cell surface and cell wall proteins. Two different approaches were used to analyze the proteins. The first was a gel-based method where 1D SDS-PAGE was used for separation of the proteins prior to reverse phase liquid chromatography tandem mass spectrometry (LC-MS/MS). The second method used MudPIT analysis (Multi dimensional Protein Identification Technique), where proteins are digested and separated using cation exchange and reversed phase separations before the MS/MS analysis (LC/LC-MS/MS). Overall, gel-based LC-MS/MS analysis resulted in more protein identifications than the MudPIT analysis. Combination of the results lead to identification of more than 1200 proteins, approximately 16% of the proteins coded from the annotated genome of Burkholderia species. Several virulence factors were detected including flagellin, porin, peroxiredoxin and zinc proteases.