Fumonisin B1 is a mycotoxin produced by fungi of the genus Fusarium, common pathogens of corn and other grain plants. Toxic effects associated with fumonisin B1 include equine leukoencephalomacia, porcine pulmonary edema, rat renal carcinoma, and murine hepatocellular carcinoma. Increased risk for esophageal cancer in humans has been epidemiologically associated with consumption of corn contaminated with Fusarium, suggesting that fumonisin B1 may be involved. The biological effects of fumonisin B1 exposure result primarily from disruption of de novo sphingolipid biosynthesis via inhibition of ceramide synthase. Exposure of animals or cultured cells to fumonisin B1 results in the characteristic accumulation of sphinganine, a toxic sphingolipid intermediate, concomitant with depletion of essential complex sphingolipids. Ceramide synthase has not been purified to homogeniety and characterized. We prepared crude ceramide synthase from detergent-extracted rat liver homogenates using PEG-precipitation and cation exchange chromatography. Ceramide synthase activity was then sequestered, using fumonisin B1 covalently coupled to POROS-NH particles, and eluted selectively. The observed 119-fold enrichment in specific activity demonstrates the utility of fumonisin-POROS affinity chromatography in the purification of ceramide synthase.
Jernigan, S.; Melchior, W. B. Jr.; Jenkins, G. R.; Rowland, K. L.; Roberts, D. W.; Howard, P. C.; and Tolleson, H.
"Characterization of a POROSTM-fumonisin B1 Affinity Column for Isolating Ceramide Synthase from Rat Liver,"
Journal of the Arkansas Academy of Science: Vol. 55
, Article 11.
Available at: http://scholarworks.uark.edu/jaas/vol55/iss1/11