Branch specificity of purified rat liver Golgi UDP-galactose: N-acetylglucosamine beta-1,4-galactosyltransferase. Preferential transfer of of galactose on the GlcNAc beta 1,2-Man alpha 1,3-branch of a complex biantennary Asn-linked oligosaccharide.
In the final stages of the terminal glycosylation of N-linked complex oligosaccharides, UDP-galactose: N-acetylglucosamine beta-1,4-galactosyltransferase (galactosyltransferase) transfers galactose (Gal) onto the N-acetylglucosamine (GlcNAc) residue of each branch of a biantennary oligosaccharide. Purified rat liver Golgi galactosyltransferase was used with GlcNAc beta 1,2-Man alpha 1,6-(GlcNAc beta 1,2-Man alpha 1,3-)-Man beta 1,4-GlcNAc beta 1,4-(Fuc alpha 1,6-)-GlcNAc-Asn in order to determine the sequence of addition of Gal residues to the biantennary oligosaccharide. The different galactosylated products were separated by concanavalin A affinity chromatography and high voltage paper electrophoresis in borate. It was found that Gal was transferred at a much faster rate to the GlcNAc beta 1,2-Man alpha 1,3-branch than to the GlcNAc beta 1,2-Man alpha 1,6-branch, i.e. k1 was at least 5 times larger than k2. Also, k3 was larger than k4, indicating that most of the digalactosylated product "GG" was formed by the sequential addition of Gal to the Man alpha 1,3-branch followed by addition to the Man alpha 1,6-branch. The preferential galactosylation of the GlcNAc beta 1,2-Man alpha 1,3-branch may explain the formation of the asymmetrical oligosaccharides found in bovine and human IgG.
D000818:Animals; D001216:Asparagine; D005690:Galactose; D006056:Golgi Apparatus / Q000201:enzymology*; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D007788:Lactose Synthase / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D009237:N-Acetyllactosamine Synthase / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity
D009844:Oligosaccharides; D001216:Asparagine; D007788:Lactose Synthase; D009237:N-Acetyllactosamine Synthase; D005690:Galactose