Brefeldin A-induced increase of sphingomyelin synthesis. Assay for the action of the antibiotic in mammalian cells.

Abstract

Brefeldin A leads to an increase of sphingomyelin in Chinese hamster ovary cells. The antibiotic is known to cause a dramatic morphological change of the endomembrane system in various mammalian cells resulting in a redistribution of Golgi resident proteins to the endoplasmic reticulum (Lippincott-Schwartz, J., Donaldson, J. G., Schweizer, A., Berger, E. G., Hauri, H. P., Yuan, L. C., and Klausner, R. D. (1990) Cell 60, 821-836). A strict correlation was found between the brefeldin A-induced increase of sphingomyelin and the biochemical criteria that apply for this morphological change. From our data we conclude that the increase in sphingomyelin caused by brefeldin A reflects translocation of the enzyme sphingomyelin synthase from the Golgi apparatus to the endoplasmic reticulum. Using a radioactively labeled truncated ceramide this increase in sphingomyelin synthesis is easily detectable, and thus this method can serve as a convenient biochemical assay for the action of brefeldin A in mammalian cells.