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%).
Dasgupta
S
S
Department of Neurology, Medical University of South Carolina, Charleston 29425.
Chien
J L
JL
Hogan
E L
EL
D000818:Animals; D001345:Autoradiography; D001665:Binding Sites / Q000187:drug effects; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D004795:Enzyme Stability; D005915:Globosides / Q000378:metabolism*; D006028:Glycosphingolipids / Q000378:metabolism*; D009132:Muscles / Q000196:embryology / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000494:pharmacology*; D012799:Sialyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D012995:Solubility; D013109:Sphingomyelins / Q000494:pharmacology*
D005915:Globosides; D006028:Glycosphingolipids; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D012799:Sialyltransferases