Towards delineating functions within the fasciola secreted cathepsin l protease family by integrating in vivo based sub-proteomics and phylogenetics.
BACKGROUND
fasciola hepatica, along with Fasciola gigantica, is the causative agent of fasciolosis, a foodborne zoonotic disease affecting grazing animals and humans worldwide. Pathology is directly related to the release of parasite proteins that facilitate establishment within the host. The dominant components of these excretory-secretory (ES) products are also the most promising vaccine candidates, the cathepsin L (Cat L) protease family.


RESULTS
the sub-proteome of Cat L proteases from adult F. hepatica ES products derived from in vitro culture and in vivo from ovine host bile were compared by 2-DE. The individual Cat L proteases were identified by tandem mass spectrometry with the support of an in-house translated liver fluke EST database. The study reveals plasticity within the CL1 clade of Cat L proteases; highlighted by the identification of a novel isoform and CL1 sub-clade, resulting in a new Cat L phylogenetic analysis including representatives from other adult Cat L phylogenetic clades. Additionally, for the first time, mass spectrometry was shown to be sufficiently sensitive to reveal single amino acid polymorphisms in a resolved 2-DE protein spot derived from pooled population samples.


CONCLUSIONS
we have investigated the sub-proteome at the population level of a vaccine target family using the Cat L proteases from F. hepatica as a case study. We have confirmed that F. hepatica exhibits more plasticity in the expression of the secreted CL1 clade of Cat L proteases at the protein level than previously realised. We recommend that superfamily based vaccine discovery programmes should screen parasite populations from different host populations and, if required, different host species via sub-proteomic assay in order to confirm the relative expression at the protein level prior to the vaccine development phase.
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, United Kingdom. rom@aber.ac.uk
Russell M
RM
Morphew
Hazel A
HA
Wright
E James
EJ
Lacourse
Joanne
J
Porter
John
J
Barrett
Debra J
DJ
Woods
Peter M
PM
Brophy
D000818:Animals; D001646:Bile / Q000469:parasitology; D056668:Cathepsin L / Q000235:genetics* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005210:Fasciola hepatica / Q000201:enzymology* / Q000302:isolation & purification; D020869:Gene Expression Profiling; D006041:Goats; D010802:Phylogeny; D040901:Proteomics
D056668:Cathepsin L
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