Substrate specificity of alpha 2----3-sialyltransferases in ganglioside biosynthesis of rat liver golgi.
The acceptor specificities of four sialytransferases (I, II, IV and V) involved in ganglioside biosynthesis were studied in Golgi vesicles derived from rat liver. The activities of these sialytransferases were strongly detergent-dependent. Competition experiments with different detergent concentrations using LacCer (Gal beta 1----4Glc beta 1----1Cer), GM1a [Gal beta 1----3GalNAc beta 1----4(NeuAc alpha 2----3)Gal beta 1----4Glc beta 1----1Cer] and GD1b [Gal beta 1----3GalNAc beta 1----4(NeuAc alpha 2----8NeuAc alpha 2----3)Gal beta 1----4Glc beta 1----1Cer] as substrates, and as mutual inhibitors for ganglioside sialyltransferase activity, suggested that sialyltransferase IV was able to catalyze the sialyltransfer in alpha 2----3 linkage to the galactose residues of LacCer as well as of GM1a and GD1b. The other three sialyltransferases (I, II and V) seemed to be quite specific for their respective glycolipid acceptors, LacCer, GM3 and GM1b, GD1a and GT1b. Furthermore the kinetic data showed that sialyltransferase I was inactive at higher detergent concentrations (greater than 75 micrograms Triton CF-54); under these conditions, formation of GM3 and GD1a was catalyzed only by sialyltransferase IV. These results have been integrated into a model for ganglioside biosynthesis and its regulation.
Iber
H
H
Institut für Organische Chemie und Biochemie der Universität Bonn, Federal Republic of Germany.
van Echten
G
G
Sandhoff
K
K
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D003902:Detergents / Q000494:pharmacology; D005732:Gangliosides / Q000096:biosynthesis*; D006028:Glycosphingolipids / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D051381:Rats; D012799:Sialyltransferases / Q000378:metabolism*; D013379:Substrate Specificity; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase
D003902:Detergents; D005732:Gangliosides; D006028:Glycosphingolipids; D012799:Sialyltransferases; D000094663:beta-Galactoside alpha-2,3-Sialyltransferase