Proteomic analyses of Ehrlichia ruminantium highlight differential expression of MAP1-family proteins.
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.
IBET, Apartado 12, 2780-901 Oeiras, Portugal. isabel_m31@hotmail.com
Isabel
I
Marcelino
André Martinho
AM
de Almeida
Catarina
C
Brito
Damien F
DF
Meyer
Mónica
M
Barreto
Christian
C
Sheikboudou
Catarina F
CF
Franco
Dominique
D
Martinez
Thierry
T
Lefrançois
Nathalie
N
Vachiéry
Manuel J T
MJ
Carrondo
Ana Varela
AV
Coelho
Paula M
PM
Alves
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002417:Cattle; D002478:Cells, Cultured; D016995:Ehrlichia ruminantium / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000382:microbiology; D006357:Heartwater Disease / Q000382:microbiology; D008969:Molecular Sequence Data; D005810:Multigene Family; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D020543:Proteome