Functional involvement of proteins, interacting with sphingolipids, in sphingolipid transport to the canalicular membrane in the human hepatocytic cell line, HepG2?

Hepatology : Official Journal of the American Association for the Study of Liver Diseases
M M ZegersD Hoekstra

Abstract

A photoreactive sphingolipid precursor was used to investigate the potential involvement of protein-lipid interactions that may convey specificity to sphingolipid transport in the human hepatoma cell line, HepG2. A 125I-labeled, photoreactive ceramide, 125I-N3-Cer, was incubated with the cells and became incorporated into two sphingolipid products. The major product was photoreactive sphingomyelin (125I-N3-SM) (25% of total radioactivity), while only minor amounts of photoreactive glucosylceramide (125I-N3-GlcCer) were formed (< 2%). After photoactivation, a restricted number of proteins was labeled. Given the absolute amounts of the newly synthesized, photoreactive lipids and their precursor present in the cells, labeling of the proteins can be assumed to be derived from interaction with either ceramide (Cer) or sphingomyelin (SM), or both. To discriminate between these possibilities, photoactivation and protein analysis was performed in cells treated with D-threo-1-phenyl-2-decanoyl amino-3-morpholino-1-propanol (PDMP), an inhibitor of sphingolipid biosynthesis. In treated cells, the radioactive SM pool was reduced by approximately 80%. Concomitantly, labeling of a 60-kd protein, seen in control cells, decreased. Furthermore,...Continue Reading

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Citations

Oct 7, 2006·Current Opinion in Gastroenterology·S J Karpen, J M Crawford

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