A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

BMC Microbiology

Research Article

Adriano MC Pimenta¹  Agenor V Santos¹  Sérgio C Oliveira¹  Luis GC Pacheco²  Thiago LP Castro³  Núbia Seyffert³  Anderson Miyoshi³  Vasco Azevedo³  Wanderson M Silva³  Anderson R Santos³  Maria AR Carvalho⁴  Simone G Santos⁴  Luiz M Farias⁴  Artur Silva⁵  Roberto Meyer⁶  Christopher G Dowson⁷  James H Scrivens⁷  Susan E Slade⁷ 

[1] Department of Biochemistry and Immunology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 31.270-901, Belo Horizonte, Brazil;Department of Biochemistry and Immunology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 31.270-901, Belo Horizonte, Brazil;Department of General Biology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 31.270-901, Belo Horizonte, Brazil;Institute of Health Sciences, Universidade Federal da Bahia, Av. Reitor Miguel Calmon, 40.110-902, Salvador, Brazil;Department of General Biology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 31.270-901, Belo Horizonte, Brazil;Department of Microbiology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 31.270-901, Belo Horizonte, Brazil;Genome and Proteome Network of the State of Pará, Universidade Federal do Pará, 66.075-110, R. Augusto Corrêa, Belém, Brazil;Institute of Health Sciences, Universidade Federal da Bahia, Av. Reitor Miguel Calmon, 40.110-902, Salvador, Brazil;School of Life Sciences, University of Warwick, Gibbet Hill Road, CV4 7AL, Coventry, United Kingdom;

关键词: Extracellular Protein;    C231 Strain;    Chemically Define Medium;    Corynebacterium Diphtheriae;    Extracellular Proteome;   

DOI  :  10.1186/1471-2180-11-12

 received in 2010-10-15, accepted in 2011-01-17,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundBacterial 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.ResultsAn 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.ConclusionsComparative 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.

【 授权许可】

Unknown   
© Pacheco et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.