A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis.
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.
Department of General Biology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av, Antônio Carlos, Belo Horizonte, 31,270-901, Brazil.
Luis G C
LG
Pacheco
Susan E
SE
Slade
Núbia
N
Seyffert
Anderson R
AR
Santos
Thiago L P
TL
Castro
Wanderson M
WM
Silva
Agenor V
AV
Santos
Simone G
SG
Santos
Luiz M
LM
Farias
Maria A R
MA
Carvalho
Adriano M C
AM
Pimenta
Roberto
R
Meyer
Artur
A
Silva
James H
JH
Scrivens
Sérgio C
SC
Oliveira
Anderson
A
Miyoshi
Christopher G
CG
Dowson
Vasco
V
Azevedo
D001426:Bacterial Proteins / Q000032:analysis*; D002853:Chromatography, Liquid; D016925:Corynebacterium pseudotuberculosis / Q000378:metabolism*; D013058:Mass Spectrometry; D040901:Proteomics / Q000379:methods*
D001426:Bacterial Proteins
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