title: Comparative proteomic analysis of triclabendazole response in the liver fluke Fasciola hepatica.
abstract: Control of Fasciola hepatica infections of livestock in the absence of vaccines depends largely on the chemical triclabendazole (TCBZ) because it is effective against immature and adult parasites. Overdependence on a single drug and improper application is considered a significant factor in increasing global reports of fluke resistant to TCBZ. The mode(s) of action and biological target(s) of TCBZ are not confirmed, delaying detection and the monitoring of early TCBZ resistance. In this study, to further understand liver fluke response to TCBZ, the soluble proteomes of TCBZ-resistant and TCBZ-susceptible isolates of F. hepatica were compared with and without in vitro exposure to the metabolically active form of the parent drug triclabendazole sulphoxide (TCBZ-SO), via two-dimensional gel electrophoresis (2-DE). Gel image analysis revealed proteins displaying altered synthesis patterns and responses both between isolates and under TCBZ-SO exposure. These proteins were identified by mass spectrometry supported by a F. hepatica expressed sequence tag (EST) data set. The TCBZ responding proteins were grouped into three categories; structural proteins, energy metabolism proteins, and "stress" response proteins. This single proteomic investigation supported the reductionist experiments from many laboratories that collectively suggest TCBZ has a range of effects on liver fluke metabolism. Proteomics highlighted differences in the innate proteome profile of different fluke isolates that may influence future therapy and diagnostics design. Two of the TCBZ responding proteins, a glutathione transferase and a fatty acid binding protein, were cloned, produced as recombinants, and both found to bind TCBZ-SO at physiologically relevant concentrations, which may indicate a role in TCBZ metabolism and resistance.
authors: Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, United Kingdom.
authors: Gustavo
authors: G
authors: Chemale
authors: Samirah
authors: S
authors: Perally
authors: E James
authors: EJ
authors: LaCourse
authors: Mark C
authors: MC
authors: Prescott
authors: Laura M
authors: LM
authors: Jones
authors: Deborah
authors: D
authors: Ward
authors: Myles
authors: M
authors: Meaney
authors: Elizabeth
authors: E
authors: Hoey
authors: Gerard P
authors: GP
authors: Brennan
authors: Ian
authors: I
authors: Fairweather
authors: Alan
authors: A
authors: Trudgett
authors: Peter M
authors: PM
authors: Brophy
mesh_terms: D000818:Animals; D000871:Anthelmintics / Q000494:pharmacology; D001562:Benzimidazoles / Q000494:pharmacology*; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism / Q000187:drug effects; D020224:Expressed Sequence Tags; D005210:Fasciola hepatica / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D015801:Helminth Proteins / Q000235:genetics / Q000378:metabolism*; D008099:Liver / Q000469:parasitology; D040901:Proteomics / Q000379:methods*; D012756:Sheep; D015398:Signal Transduction / Q000187:drug effects; D013552:Swine; D053719:Tandem Mass Spectrometry; D000077682:Triclabendazole
chemical_list: D000871:Anthelmintics; D001562:Benzimidazoles; D015801:Helminth Proteins; D000077682:Triclabendazole