Brefeldin A-induced increase of sphingomyelin synthesis. Assay for the action of the antibiotic in mammalian cells.
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.
Brüning
A
A
Institut für Biochemie I, Universität Heidelberg, Germany.
Karrenbauer
A
A
Schnabel
E
E
Wieland
F T
FT
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D020126:Brefeldin A; D016466:CHO Cells; D002460:Cell Line; D002794:Choline / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D006224:Cricetinae; D003517:Cyclopentanes / Q000494:pharmacology*; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D007700:Kinetics; D011865:Radioisotope Dilution Technique; D013109:Sphingomyelins / Q000096:biosynthesis*; D014316:Tritium
D000900:Anti-Bacterial Agents; D003517:Cyclopentanes; D013109:Sphingomyelins; D014316:Tritium; D005576:Colforsin; D020126:Brefeldin A; D002794:Choline