Exposure of intestinal explants to NX, but not to DON, enriches the secretome in mitochondrial proteins

Abstract

NX is a type A trichothecene produced by Fusarium graminearum with limited information on its toxicity. NX is structurally similar to deoxynivalenol (DON), only differing by the lacking keto group at C8. Because of the structural similarity of the two toxins as well as their potential co-occurrence in food and feed, it is of interest to determine the toxicity of this new compound. In this study, we compared the protein composition of the extracellular media of pig intestinal explants (secretome) exposed to 10 µM of DON or NX for 4 h compared with controls. The combination of two complementary quantitative proteomic approaches (a gel-based and a gel-free approach) identified 18 and 23 differentially abundant proteins (DAPs) for DON and NX, respectively, compared to controls. Functional analysis suggested that, whereas DON toxicity was associated with decreased cell viability and cell destruction, NX toxicity was associated with an enrichment of mitochondrial proteins in the secretome. The presence of these proteins may be associated with the already known ability of NX to induce an intestinal inflammation. Overall, our results indicated that DON- and NX-induced changes in the extracellular proteome of intestinal explants are different. The increased leakage/secretion of mitochondrial proteins by NX may be a feature of NX toxicity.

Application of 2-D DIGE to study the effect of ageing on horse meat myofibrillar sub-proteome

“…A total amount of 10 or 20 μg of extracts were loaded per lane, depending of the targeted protein, for optimal visualization of main protein and breakdown products, respectively (for details see the respective figure legends). The procedure was similar to previously published protocols [27]. Samples were separated in home-made 10-15% SDS-PAGE gradient gels under reducing and denaturing conditions and proteins then were semi-dry blotted onto NC (GE Healthcare).…”