Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach.
Neospora caninum is an important veterinary pathogen that causes abortion in cattle and neuromuscular disease in dogs. Neospora has also generated substantial interest because it is an extremely close relative of the human pathogen Toxoplasma gondii, yet does not appear to infect humans. While for T...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2011-04-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3070720?pdf=render |
| _version_ | 1818957742489993216 |
|---|---|
| author | Catherine S Sohn Tim T Cheng Michael L Drummond Eric D Peng Sarah J Vermont Dong Xia Stephen J Cheng Jonathan M Wastling Peter J Bradley |
| author_facet | Catherine S Sohn Tim T Cheng Michael L Drummond Eric D Peng Sarah J Vermont Dong Xia Stephen J Cheng Jonathan M Wastling Peter J Bradley |
| author_sort | Catherine S Sohn |
| collection | DOAJ |
| description | Neospora caninum is an important veterinary pathogen that causes abortion in cattle and neuromuscular disease in dogs. Neospora has also generated substantial interest because it is an extremely close relative of the human pathogen Toxoplasma gondii, yet does not appear to infect humans. While for Toxoplasma there are a wide array of molecular tools and reagents available for experimental investigation, relatively few reagents exist for Neospora. To investigate the unique biological features of this parasite and exploit the recent sequencing of its genome, we have used an organelle isolation and monoclonal antibody approach to identify novel organellar proteins and develop a wide array of probes for subcellular localization. We raised a panel of forty-six monoclonal antibodies that detect proteins from the rhoptries, micronemes, dense granules, inner membrane complex, apicoplast, mitochondrion and parasite surface. A subset of the proteins was identified by immunoprecipitation and mass spectrometry and reveal that we have identified and localized many of the key proteins involved in invasion and host interaction in Neospora. In addition, we identified novel secretory proteins not previously studied in any apicomplexan parasite. Thus, this organellar monoclonal antibody approach not only greatly enhances the tools available for Neospora cell biology, but also identifies novel components of the unique biological characteristics of this important veterinary pathogen. |
| first_indexed | 2024-12-20T11:14:41Z |
| format | Article |
| id | doaj.art-4758658ab4b74242b65eeb41995485cb |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T11:14:41Z |
| publishDate | 2011-04-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-4758658ab4b74242b65eeb41995485cb2022-12-21T19:42:39ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-04-0164e1838310.1371/journal.pone.0018383Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach.Catherine S SohnTim T ChengMichael L DrummondEric D PengSarah J VermontDong XiaStephen J ChengJonathan M WastlingPeter J BradleyNeospora caninum is an important veterinary pathogen that causes abortion in cattle and neuromuscular disease in dogs. Neospora has also generated substantial interest because it is an extremely close relative of the human pathogen Toxoplasma gondii, yet does not appear to infect humans. While for Toxoplasma there are a wide array of molecular tools and reagents available for experimental investigation, relatively few reagents exist for Neospora. To investigate the unique biological features of this parasite and exploit the recent sequencing of its genome, we have used an organelle isolation and monoclonal antibody approach to identify novel organellar proteins and develop a wide array of probes for subcellular localization. We raised a panel of forty-six monoclonal antibodies that detect proteins from the rhoptries, micronemes, dense granules, inner membrane complex, apicoplast, mitochondrion and parasite surface. A subset of the proteins was identified by immunoprecipitation and mass spectrometry and reveal that we have identified and localized many of the key proteins involved in invasion and host interaction in Neospora. In addition, we identified novel secretory proteins not previously studied in any apicomplexan parasite. Thus, this organellar monoclonal antibody approach not only greatly enhances the tools available for Neospora cell biology, but also identifies novel components of the unique biological characteristics of this important veterinary pathogen.http://europepmc.org/articles/PMC3070720?pdf=render |
| spellingShingle | Catherine S Sohn Tim T Cheng Michael L Drummond Eric D Peng Sarah J Vermont Dong Xia Stephen J Cheng Jonathan M Wastling Peter J Bradley Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach. PLoS ONE |
| title | Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach. |
| title_full | Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach. |
| title_fullStr | Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach. |
| title_full_unstemmed | Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach. |
| title_short | Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach. |
| title_sort | identification of novel proteins in neospora caninum using an organelle purification and monoclonal antibody approach |
| url | http://europepmc.org/articles/PMC3070720?pdf=render |
| work_keys_str_mv | AT catherinessohn identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT timtcheng identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT michaelldrummond identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT ericdpeng identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT sarahjvermont identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT dongxia identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT stephenjcheng identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT jonathanmwastling identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach AT peterjbradley identificationofnovelproteinsinneosporacaninumusinganorganellepurificationandmonoclonalantibodyapproach |