Proteomic analysis of embryonic Fasciola hepatica: Characterization and antigenic potential of a developmentally regulated heat shock protein

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dc.contributor.author Moxon, J. V.
dc.contributor.author LaCourse, E. James
dc.contributor.author Wright, H. A.
dc.contributor.author Perally, S.
dc.contributor.author Prescott, Mark C.
dc.contributor.author Gillard, Jennifer L.
dc.contributor.author Barrett, J.
dc.contributor.author Hamilton, J. V.
dc.contributor.author Brophy, P. M.
dc.date.accessioned 2011-06-09T09:32:39Z
dc.date.available 2011-06-09T09:32:39Z
dc.date.issued 2010-04-19
dc.identifier.citation Moxon , J V , LaCourse , E J , Wright , H A , Perally , S , Prescott , M C , Gillard , J L , Barrett , J , Hamilton , J V & Brophy , P M 2010 , ' Proteomic analysis of embryonic Fasciola hepatica: Characterization and antigenic potential of a developmentally regulated heat shock protein ' Veterinary Parasitology , vol 169 , no. 1-2 , pp. 62-75 . en
dc.identifier.issn 0304-4017
dc.identifier.other PURE: 165111
dc.identifier.other dspace: 2160/7018
dc.identifier.uri http://hdl.handle.net/2160/7018
dc.description Moxon, J. V., LaCourse, E. J., Wright, H. A., Perally, S., Prescott, M. C., Gillard, J. L., Barrett, J., Hamilton, J. V., Brophy, P. M. (2010). Proteomic analysis of embryonic Fasciola hepatica: Characterization and antigenic potential of a developmentally regulated heat shock protein.   Veterinary Parasitology, 169 (1-2), 62-75. Sponsorship: NERC UK; EU DeLiver project; BBSRC UK Proteomics and Cell Function Initiative (BBC503638) IMPF: 02.33 en
dc.description.abstract Fasciola hepatica is responsible for human disease and economic livestock loss on a global scale. We report the first post-genomic investigation of cellular proteins expressed by embryonic F. hepatica via two-dimensional electrophoresis, image analysis and tandem mass spectrometry. Antioxidant proteins and protein chaperones are prominently expressed by embryonic F. hepatica. Molecular differences between the egg and other characterized F. hepatica lifecycle stages were noted. Furthermore, proteins expressed within liver fluke eggs differ to those isolated from the well-characterized eggs of the human blood flatworm Schistosoma mansoni were revealed. Plasticity in expression of major proteins, particularly a prominently expressed 65 kDa protein cluster was seen between natural populations of embryonating F. hepatica eggs suggesting that liver fluke embryogenisis is a plastic process. Immunoblotting revealed that the abundant 65 kDa protein cluster is recognised by infection sera from three F. hepatica challenged host species. Mass spectrometry and BLAST analyses demonstrated that the 65 kDa antigen shows homology to egg antigens of other flatworm parasites, and is represented in a F. hepatica EST database constructed from adult fluke transcripts. EST clones encoding the egg antigen were re-sequenced, predicting two forms of the protein. Four clones predict a 312 aa polypeptide, three clones encode a putative 110 amino acid extension at the N-terminus which may be involved in protein secretion, although this extension was not expressed by natively extracted proteins. Consistent expression of alpha crystallin domains confirmed the protein to be a member of the alpha crystallin containing small heat shock protein (AC/sHSP) superfamily. AC/sHSPs are ubiquitous in nature, however, this is the first time a member of this protein superfamily has been described from F. hepatica. The antigenic AC/sHSP was named Fh-HSP35alpha based on predictions of molecular weight. Production of recombinant Fh-HSP35alpha reveals considerable mass discrepancy between native and recombinant proteins, although descriptions of other characterized flatworm AC/sHSPs, suggest that the native form is a dimer. Immunoblot analyses confirm that the recombinant protein is recognised by F. hepatica challenged hosts, but does not react with sera from non-infected animals. We discuss the potential of recombinant Fh-HSP35alpha as an egg-based diagnostic marker for liver fluke infection. en
dc.format.extent 14 en
dc.language.iso eng
dc.relation.ispartof Veterinary Parasitology en
dc.subject flatworm en
dc.subject embryo en
dc.subject heat shock protein en
dc.subject egg en
dc.subject antigen en
dc.subject embryogenesis en
dc.title Proteomic analysis of embryonic Fasciola hepatica: Characterization and antigenic potential of a developmentally regulated heat shock protein en
dc.type Text en
dc.type.publicationtype Article (Journal) en
dc.identifier.doi http://dx.doi.org/10.1016/j.vetpar.2009.12.031
dc.contributor.institution Institute of Biological, Environmental and Rural Sciences en
dc.description.status Peer reviewed en


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