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Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach

Identification of novel proteins in Neospora caninum using an organelle purification and monoclonal antibody approach Thumbnail


Abstract

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.

Acceptance Date Feb 28, 2011
Publication Date Apr 4, 2011
Publicly Available Date Mar 29, 2024
Journal PLoS One
Print ISSN 1932-6203
Publisher Public Library of Science
Pages e18383 -?
DOI https://doi.org/10.1371/journal.pone.0018383
Keywords Toxoplasma, Immunoprecipitation, Cellular structures and organelles, Membrane proteins, Membrane staining, Vacuoles, Antibodies, Toxoplasma gondii
Publisher URL http://dx.doi.org/10.1371/journal.pone.0018383

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