Biomarkers for ragwort poisoning in horses: identification of protein targets.
BACKGROUND
Ingestion of the poisonous weed ragwort (Senecio jacobea) by horses leads to irreversible liver damage. The principal toxins of ragwort are the pyrrolizidine alkaloids that are rapidly metabolised to highly reactive and cytotoxic pyrroles, which can escape into the circulation and bind to proteins. In this study a non-invasive in vitro model system has been developed to investigate whether pyrrole toxins induce specific modifications of equine blood proteins that are detectable by proteomic methods.


RESULTS
One dimensional gel electrophoresis revealed a significant alteration in the equine plasma protein profile following pyrrole exposure and the formation of a high molecular weight protein aggregate. Using mass spectrometry and confirmation by western blotting the major components of this aggregate were identified as fibrinogen, serum albumin and transferrin.


CONCLUSIONS
These findings demonstrate that pyrrolic metabolites can modify equine plasma proteins. The high molecular weight aggregate may result from extensive inter- and intra-molecular cross-linking of fibrinogen with the pyrrole. This model has the potential to form the basis of a novel proteomic strategy aimed at identifying surrogate protein biomarkers of ragwort exposure in horses and other livestock.
Moore
Rowan E
RE
Proteomics and Functional Genomics Research Group, Faculty of Veterinary Science, University of Liverpool, Crown Street, Liverpool, L69 7ZJ, UK. r.e.moore@liverpool.ac.uk
Knottenbelt
Derek
D
Matthews
Jacqueline B
JB
Beynon
Robert J
RJ
Whitfield
Phillip D
PD
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism*; D015153:Blotting, Western; D005340:Fibrinogen / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism; D006734:Horse Diseases / Q000175:diagnosis* / Q000209:etiology*; D006736:Horses / Q000097:blood / Q000378:metabolism*; D010939:Plant Poisoning / Q000175:diagnosis / Q000209:etiology / Q000662:veterinary*; D010947:Plants, Toxic; D040901:Proteomics; D011758:Pyrroles / Q000506:poisoning; D011763:Pyrrolizidine Alkaloids / Q000506:poisoning; D012674:Senecio / Q000506:poisoning*; D013997:Time Factors
D015415:Biomarkers; D001798:Blood Proteins; D006454:Hemoglobins; D011758:Pyrroles; D011763:Pyrrolizidine Alkaloids; D005340:Fibrinogen