Identification of a novel UDP-Glc:GlcNAc beta1-->4-glucosyltransferase in Lymnaea stagnalis that may be involved in the synthesis of complex-type oligosaccharide chains.
Several studies suggest, that the snail Lymnaea stagnalis contains glycoproteins whose oligosaccharide side chains have structural features not commonly found in mammalian glycoproteins. In this study, prostate glands of L. stagnalis were incubated in media containing either [(3)H]-mannose, [(3)H]-glucosamine, or [(3)H]-galactose, and the metabolically radiolabeled protein-bound oligosaccharides were analyzed. The newly synthesized diantennary-like complex-type asparagine-linked chains contained a considerable amount of glucose, next to mannose, GlcNAc, fucose, galactose, and traces of GalNAc. Since glucose has not been found before as a constituent of diantennary N-linked glycans as far as we know, we assayed the prostate gland of L. stagnalis for a potential glucosyltransferase activity involved in the biosynthesis of such structures. We report here, that the prostate gland of L. stagnalis contains a beta1-->4-glucosyltransferase activity that transfers glucose from UDP-glucose to acceptor substrates carrying a terminal N-acetylglucosamine. The enzyme prefers substrates carrying a terminal GlcNAc that is beta6 linked to a Gal or a GalNAc, structures occurring in O-linked glycans, or a GlcNAc that is beta2 linked to mannose, as is present in N-linked glycans. Based on combined structural and enzymatic data, we propose that the novel beta1-->4-gluco-syltransferase present in the prostate gland may be involved in the biosynthesis of Glcbeta1-->4GlcNAc units in complex-type glycans, in particular in N-linked diantennary glycans.
van Die
I
I
Department of Medical Chemistry, Vrije Universiteit, Van der Boechorststraat 7, 1081BT Amsterdam, The Netherlands.
Cummings
R D
RD
van Tetering
A
A
Hokke
C H
CH
Koeleman
C A
CA
van den Eijnden
D H
DH
D000818:Animals; D002240:Carbohydrate Sequence; D002847:Chromatography, Agarose; D003208:Concanavalin A / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005470:Fluorometry; D005690:Galactose / Q000378:metabolism; D005964:Glucosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification; D008195:Lymnaea / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D011134:Polysaccharides / Q000096:biosynthesis / Q000378:metabolism; D011467:Prostate / Q000201:enzymology* / Q000378:metabolism; D013379:Substrate Specificity; D014316:Tritium
D006023:Glycoproteins; D009844:Oligosaccharides; D011134:Polysaccharides; D014316:Tritium; D003208:Concanavalin A; D005964:Glucosyltransferases; D005690:Galactose