TY - JOUR
T1 - Proteomic analyses of Ehrlichia ruminantium highlight differential expression of MAP1-family proteins
AU - Marcelino, Isabel
AU - de Almeida, André Martinho
AU - Brito, Catarina
AU - Meyer, Damien F.
AU - Barreto, Mónica
AU - Sheikboudou, Christian
AU - Franco, Catarina F.
AU - Martinez, Dominique
AU - Lefrançois, Thierry
AU - Vachiéry, Nathalie
AU - Carrondo, Manuel J.T.
AU - Coelho, Ana Varela
AU - Alves, Paula M.
N1 - Funding Information:
The authors acknowledge the financial support received from the European Commission (contract number FP6-003713) and the Fundação para a Ciência e Tecnologia (PTDC/CVT/114118/2009). Authors André M. Almeida, Catarina Brito and Catarina F. Franco acknowledge financial support from the Ciência 2007/2008 programs (A.M. Almeida and C. Brito) and PhD grant SFRH/BD/29799/2006 (CF Franco) all from FCT/MCTES (Lisbon, Portugal) . We would like to thank Rita Francisco for collaboration in gel analysis, Yane Kandassamy for technical assistance in gene cloning and Ken Giraud-Girard for animal handling during polyclonal antibody production.
Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.
PY - 2012/5/4
Y1 - 2012/5/4
N2 - The Rickettsiales Ehrlichia ruminantium (ER) is the causative agent of heartwater, a fatal tick-borne disease of livestock in sub-Saharan Africa and in the Caribbean, posing strong economical constraints to livestock production. In an attempt to identify the most prominent proteins expressed by this bacterium, especially those encoded by the major antigenic protein 1 (map1) multigene family, a proteome map of ER cultivated in endothelial cells was constructed by using two dimensional gel electrophoresis combined with mass spectrometry. Among the sixty-four spots detected, we could identify only four proteins from the MAP1-family; the other proteins detected were mainly related to energy, amino acid and general metabolism (26%), to protein turnover, chaperones and survival (21%) and to information processes (14%) or classified as hypothetical proteins (23%).Additional studies on MAP1-family protein using immunochemical labeling also revealed that these proteins are differentially expressed along the bacterium life cycle, presenting different structural organization. Interestingly, when infectious elementary bodies (EBs) are released from host cells, MAP1 appears to be organized in SDS and heat-resistant dimers and trimers stabilized by disulfide bridges.Overall, the results presented herein not only reveal the first partial proteome map of ER but provide new insights on the expression ER MAP1-family proteins in host endothelial cells.
AB - The Rickettsiales Ehrlichia ruminantium (ER) is the causative agent of heartwater, a fatal tick-borne disease of livestock in sub-Saharan Africa and in the Caribbean, posing strong economical constraints to livestock production. In an attempt to identify the most prominent proteins expressed by this bacterium, especially those encoded by the major antigenic protein 1 (map1) multigene family, a proteome map of ER cultivated in endothelial cells was constructed by using two dimensional gel electrophoresis combined with mass spectrometry. Among the sixty-four spots detected, we could identify only four proteins from the MAP1-family; the other proteins detected were mainly related to energy, amino acid and general metabolism (26%), to protein turnover, chaperones and survival (21%) and to information processes (14%) or classified as hypothetical proteins (23%).Additional studies on MAP1-family protein using immunochemical labeling also revealed that these proteins are differentially expressed along the bacterium life cycle, presenting different structural organization. Interestingly, when infectious elementary bodies (EBs) are released from host cells, MAP1 appears to be organized in SDS and heat-resistant dimers and trimers stabilized by disulfide bridges.Overall, the results presented herein not only reveal the first partial proteome map of ER but provide new insights on the expression ER MAP1-family proteins in host endothelial cells.
KW - Ehrlichia ruminantium
KW - MAP1-family proteins
KW - Mass spectrometry
KW - Proteome
KW - Two-dimensional electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=84858746865&partnerID=8YFLogxK
U2 - 10.1016/j.vetmic.2011.11.022
DO - 10.1016/j.vetmic.2011.11.022
M3 - Article
C2 - 22204792
VL - 156
SP - 305
EP - 314
JO - Veterinary Microbiology
JF - Veterinary Microbiology
SN - 0378-1135
IS - 3-4
ER -