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Open Access Highly Accessed Research article

A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

Luis GC Pacheco123, Susan E Slade4, Núbia Seyffert2, Anderson R Santos2, Thiago LP Castro2, Wanderson M Silva2, Agenor V Santos1, Simone G Santos5, Luiz M Farias5, Maria AR Carvalho5, Adriano MC Pimenta1, Roberto Meyer3, Artur Silva6, James H Scrivens4, Sérgio C Oliveira1, Anderson Miyoshi2, Christopher G Dowson4 and Vasco Azevedo2*

Author affiliations

1 Department of Biochemistry and Immunology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, Belo Horizonte, 31.270-901, Brazil

2 Department of General Biology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, Belo Horizonte, 31.270-901, Brazil

3 Institute of Health Sciences, Universidade Federal da Bahia, Av. Reitor Miguel Calmon, Salvador, 40.110-902, Brazil

4 School of Life Sciences, University of Warwick, Gibbet Hill Road, Coventry, CV4 7AL, United Kingdom

5 Department of Microbiology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, Belo Horizonte, 31.270-901, Brazil

6 Genome and Proteome Network of the State of Pará, Universidade Federal do Pará, R. Augusto Corrêa, Belém, 66.075-110, Brazil

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Citation and License

BMC Microbiology 2011, 11:12  doi:10.1186/1471-2180-11-12

Published: 17 January 2011

Abstract

Background

Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA) in sheep and goats.

Results

An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MSE) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins.

Conclusions

Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far.