Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium
The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium , the obligate intracellular bacterium that causes heart water. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia's OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LC-MALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp . The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins. We identified proteins potentially involved in E. ruminantium pathogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium.
id |
dig-cirad-fr-575545 |
---|---|
record_format |
koha |
institution |
CIRAD FR |
collection |
DSpace |
country |
Francia |
countrycode |
FR |
component |
Bibliográfico |
access |
En linea |
databasecode |
dig-cirad-fr |
tag |
biblioteca |
region |
Europa del Oeste |
libraryname |
Biblioteca del CIRAD Francia |
language |
eng |
topic |
L73 - Maladies des animaux U30 - Méthodes de recherche Ehrlichia protéine bactérienne électrophorèse sur gel chromatographie sur gel spectrométrie de masse bactériologie vaccin maladie transmissible par tiques pathogénèse agent pathogène virulence pouvoir pathogène Ehrlichia ruminantium http://aims.fao.org/aos/agrovoc/c_2504 http://aims.fao.org/aos/agrovoc/c_24141 http://aims.fao.org/aos/agrovoc/c_34949 http://aims.fao.org/aos/agrovoc/c_28294 http://aims.fao.org/aos/agrovoc/c_12421 http://aims.fao.org/aos/agrovoc/c_768 http://aims.fao.org/aos/agrovoc/c_8130 http://aims.fao.org/aos/agrovoc/c_24908 http://aims.fao.org/aos/agrovoc/c_24881 http://aims.fao.org/aos/agrovoc/c_5630 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_34694 http://aims.fao.org/aos/agrovoc/c_3406 http://aims.fao.org/aos/agrovoc/c_3081 L73 - Maladies des animaux U30 - Méthodes de recherche Ehrlichia protéine bactérienne électrophorèse sur gel chromatographie sur gel spectrométrie de masse bactériologie vaccin maladie transmissible par tiques pathogénèse agent pathogène virulence pouvoir pathogène Ehrlichia ruminantium http://aims.fao.org/aos/agrovoc/c_2504 http://aims.fao.org/aos/agrovoc/c_24141 http://aims.fao.org/aos/agrovoc/c_34949 http://aims.fao.org/aos/agrovoc/c_28294 http://aims.fao.org/aos/agrovoc/c_12421 http://aims.fao.org/aos/agrovoc/c_768 http://aims.fao.org/aos/agrovoc/c_8130 http://aims.fao.org/aos/agrovoc/c_24908 http://aims.fao.org/aos/agrovoc/c_24881 http://aims.fao.org/aos/agrovoc/c_5630 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_34694 http://aims.fao.org/aos/agrovoc/c_3406 http://aims.fao.org/aos/agrovoc/c_3081 |
spellingShingle |
L73 - Maladies des animaux U30 - Méthodes de recherche Ehrlichia protéine bactérienne électrophorèse sur gel chromatographie sur gel spectrométrie de masse bactériologie vaccin maladie transmissible par tiques pathogénèse agent pathogène virulence pouvoir pathogène Ehrlichia ruminantium http://aims.fao.org/aos/agrovoc/c_2504 http://aims.fao.org/aos/agrovoc/c_24141 http://aims.fao.org/aos/agrovoc/c_34949 http://aims.fao.org/aos/agrovoc/c_28294 http://aims.fao.org/aos/agrovoc/c_12421 http://aims.fao.org/aos/agrovoc/c_768 http://aims.fao.org/aos/agrovoc/c_8130 http://aims.fao.org/aos/agrovoc/c_24908 http://aims.fao.org/aos/agrovoc/c_24881 http://aims.fao.org/aos/agrovoc/c_5630 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_34694 http://aims.fao.org/aos/agrovoc/c_3406 http://aims.fao.org/aos/agrovoc/c_3081 L73 - Maladies des animaux U30 - Méthodes de recherche Ehrlichia protéine bactérienne électrophorèse sur gel chromatographie sur gel spectrométrie de masse bactériologie vaccin maladie transmissible par tiques pathogénèse agent pathogène virulence pouvoir pathogène Ehrlichia ruminantium http://aims.fao.org/aos/agrovoc/c_2504 http://aims.fao.org/aos/agrovoc/c_24141 http://aims.fao.org/aos/agrovoc/c_34949 http://aims.fao.org/aos/agrovoc/c_28294 http://aims.fao.org/aos/agrovoc/c_12421 http://aims.fao.org/aos/agrovoc/c_768 http://aims.fao.org/aos/agrovoc/c_8130 http://aims.fao.org/aos/agrovoc/c_24908 http://aims.fao.org/aos/agrovoc/c_24881 http://aims.fao.org/aos/agrovoc/c_5630 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_34694 http://aims.fao.org/aos/agrovoc/c_3406 http://aims.fao.org/aos/agrovoc/c_3081 Moumène, Amal Marcelino, Isabel Ventosa, Miguel Gros, Olivier Lefrançois, Thierry Vachiery, Nathalie Meyer, Damien Coelho, Ana Varela Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium |
description |
The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium , the obligate intracellular bacterium that causes heart water. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia's OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LC-MALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp . The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins. We identified proteins potentially involved in E. ruminantium pathogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium. |
format |
article |
topic_facet |
L73 - Maladies des animaux U30 - Méthodes de recherche Ehrlichia protéine bactérienne électrophorèse sur gel chromatographie sur gel spectrométrie de masse bactériologie vaccin maladie transmissible par tiques pathogénèse agent pathogène virulence pouvoir pathogène Ehrlichia ruminantium http://aims.fao.org/aos/agrovoc/c_2504 http://aims.fao.org/aos/agrovoc/c_24141 http://aims.fao.org/aos/agrovoc/c_34949 http://aims.fao.org/aos/agrovoc/c_28294 http://aims.fao.org/aos/agrovoc/c_12421 http://aims.fao.org/aos/agrovoc/c_768 http://aims.fao.org/aos/agrovoc/c_8130 http://aims.fao.org/aos/agrovoc/c_24908 http://aims.fao.org/aos/agrovoc/c_24881 http://aims.fao.org/aos/agrovoc/c_5630 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_34694 http://aims.fao.org/aos/agrovoc/c_3406 http://aims.fao.org/aos/agrovoc/c_3081 |
author |
Moumène, Amal Marcelino, Isabel Ventosa, Miguel Gros, Olivier Lefrançois, Thierry Vachiery, Nathalie Meyer, Damien Coelho, Ana Varela |
author_facet |
Moumène, Amal Marcelino, Isabel Ventosa, Miguel Gros, Olivier Lefrançois, Thierry Vachiery, Nathalie Meyer, Damien Coelho, Ana Varela |
author_sort |
Moumène, Amal |
title |
Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium |
title_short |
Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium |
title_full |
Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium |
title_fullStr |
Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium |
title_full_unstemmed |
Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium |
title_sort |
proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen ehrlichia ruminantium |
url |
http://agritrop.cirad.fr/575545/ http://agritrop.cirad.fr/575545/1/document_575545.pdf |
work_keys_str_mv |
AT moumeneamal proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT marcelinoisabel proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT ventosamiguel proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT grosolivier proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT lefrancoisthierry proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT vachierynathalie proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT meyerdamien proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium AT coelhoanavarela proteomicprofilingoftheoutermembranefractionoftheobligateintracellularbacterialpathogenehrlichiaruminantium |
_version_ |
1792498794137387008 |
spelling |
dig-cirad-fr-5755452024-01-28T22:35:02Z http://agritrop.cirad.fr/575545/ http://agritrop.cirad.fr/575545/ Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium. Moumène Amal, Marcelino Isabel, Ventosa Miguel, Gros Olivier, Lefrançois Thierry, Vachiery Nathalie, Meyer Damien, Coelho Ana Varela. 2015. PloS One, 10 (2):e0116758, 20 p.https://doi.org/10.1371/journal.pone.0116758 <https://doi.org/10.1371/journal.pone.0116758> Proteomic profiling of the outer membrane fraction of the obligate intracellular bacterial pathogen Ehrlichia ruminantium Moumène, Amal Marcelino, Isabel Ventosa, Miguel Gros, Olivier Lefrançois, Thierry Vachiery, Nathalie Meyer, Damien Coelho, Ana Varela eng 2015 PloS One L73 - Maladies des animaux U30 - Méthodes de recherche Ehrlichia protéine bactérienne électrophorèse sur gel chromatographie sur gel spectrométrie de masse bactériologie vaccin maladie transmissible par tiques pathogénèse agent pathogène virulence pouvoir pathogène Ehrlichia ruminantium http://aims.fao.org/aos/agrovoc/c_2504 http://aims.fao.org/aos/agrovoc/c_24141 http://aims.fao.org/aos/agrovoc/c_34949 http://aims.fao.org/aos/agrovoc/c_28294 http://aims.fao.org/aos/agrovoc/c_12421 http://aims.fao.org/aos/agrovoc/c_768 http://aims.fao.org/aos/agrovoc/c_8130 http://aims.fao.org/aos/agrovoc/c_24908 http://aims.fao.org/aos/agrovoc/c_24881 http://aims.fao.org/aos/agrovoc/c_5630 http://aims.fao.org/aos/agrovoc/c_8261 http://aims.fao.org/aos/agrovoc/c_5629 http://aims.fao.org/aos/agrovoc/c_34694 Guadeloupe France http://aims.fao.org/aos/agrovoc/c_3406 http://aims.fao.org/aos/agrovoc/c_3081 The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium , the obligate intracellular bacterium that causes heart water. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia's OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LC-MALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp . The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins. We identified proteins potentially involved in E. ruminantium pathogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium. article info:eu-repo/semantics/article Journal Article info:eu-repo/semantics/publishedVersion http://agritrop.cirad.fr/575545/1/document_575545.pdf application/pdf Cirad license info:eu-repo/semantics/openAccess https://agritrop.cirad.fr/mention_legale.html https://doi.org/10.1371/journal.pone.0116758 10.1371/journal.pone.0116758 info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0116758 info:eu-repo/semantics/altIdentifier/purl/https://doi.org/10.1371/journal.pone.0116758 |