Sialylation of lacto-N-neotetraosyl ceramide by a solubilized sialyltransferase(s) from chicken skeletal muscle: effect of phosphatidylcholine and sphingomyelin.

The sialytransferase(s) that transfers sialic acid to lacto-N-neotetraosylceramide and other glycosphingolipids with a galactose nonreducing terminus has been successfully solubilized from embryonic chicken skeletal muscle. The enzyme can be stored in 50 mM HEPES (pH 6.8), 1% Triton CF-54, and 20% glycerol at -70 degrees C for as long as six months. Addition of phosphatidylcholine or sphingomyelin (0.167%) readily reactivates the stored inactive enzymes and such activity persists for about two weeks at 0 degrees-4 degrees C with the peak activity occurring at 1 to 2 days. Sphingomyelin from chicken muscle, which contains mainly C16:0 and C18:0, is 2.1-fold more effective than bovine brain sphingomyelin at the same concentration (0.4%).