Terminal glycosylation in rat hepatic Golgi fractions: heterogeneous locations for sialic acid and galactose acceptors and their transferases.
Endogenous acceptors for N-acetylglucosamine (GlcNAc), galactose (Gal) or sialic acid (NeuAc) transfer were labeled to high activities when purified hepatic Golgi fractions were incubated with the corresponding radiolabeled nucleotide sugar in the absence of detergent. The in vitro conditions which were optimal for the endogenous glycosylation of GlcNAc and Gal acceptors (Mn2+, ATP) also promoted fusion within a subset of Golgi membranes. Electron microscope radioautography revealed that the majority of NeuAc acceptors were associated with unfused Golgi membranes, whereas the majority of Gal acceptors were localized to fused membranes. GlcNAc acceptors were approximately equally distributed between fused and unfused membranes. Under conditions in which Golgi membrane fusion was absent (-Mn2+), only NeuAc transfer was active. The majority of endogenous NeuAc acceptors were consequently assigned to the more trans regions of the hepatic Golgi apparatus as concluded from a combination of radioautography (NeuAc transfer) and acid NADPase cytochemistry (reactive medial and trans Golgi saccules). The distribution of NeuAc and Gal transferases was assessed after Percoll gradient centrifugation of disrupted Golgi fractions. The median density of NeuAc transferase was lower than that of Gal transferase. The studies are indicative of distinct Golgi components harboring the majority of acceptors and enzymes for terminal glycosylation.
Bergeron
J J
JJ
Paiement
J
J
Khan
M N
MN
Smith
C E
CE
D000117:Acetylglucosamine / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid / Q000378:metabolism; D005690:Galactose / Q000378:metabolism*; D005700:Galactosyltransferases / Q000378:metabolism*; D005944:Glucosamine / Q000031:analogs & derivatives*; D005964:Glucosyltransferases / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D008099:Liver / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D017351:N-Acetylglucosaminyltransferases; D019158:N-Acetylneuraminic Acid; D051381:Rats; D012794:Sialic Acids / Q000378:metabolism*; D012799:Sialyltransferases / Q000378:metabolism*; D014166:Transferases / Q000378:metabolism*; D014531:Uridine Diphosphate Galactose / Q000378:metabolism; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism
D012794:Sialic Acids; D014531:Uridine Diphosphate Galactose; D003569:Cytidine Monophosphate N-Acetylneuraminic Acid; D014537:Uridine Diphosphate N-Acetylglucosamine; D014166:Transferases; D005700:Galactosyltransferases; D005964:Glucosyltransferases; D017351:N-Acetylglucosaminyltransferases; C037649:N-acetyllactosaminide beta-1,6-N-acetylglucosaminyltransferase; D012799:Sialyltransferases; D019158:N-Acetylneuraminic Acid; D005944:Glucosamine; D000117:Acetylglucosamine; D005690:Galactose