Transient methylation of dolichyl oligosaccharides is an obligatory step in halobacterial sulfated glycoprotein biosynthesis.
Biosynthesis of sulfated saccharides that are linked to asparagine residues in the cell surface glycoprotein of Halobacterium halobium via a glucose residue involves sulfated dolichyl-monophosphoryl oligosaccharide intermediates (Lechner, J., Wieland, F., and Sumper, M. (1985) J. Biol. Chem. 260, 860-866). During isolation and characterization of these lipid oligosaccharides we detected a group of related compounds containing additional unidentified sugar residues. Here we report that: 1) the unknown sugar residues were 3-O-methylglucose, linked peripherally to the lipid-saccharide intermediates; 2) the 3-O-methylglucose residues in the oligosaccharides occur only at the lipid-linked level but are absent at the protein-linked level; 3) cell surface glycoprotein biosynthesis in Halobacteria in vivo is drastically depressed when S-adenosylmethionine-dependent methylation is inhibited, indicating that methylation is an obligatory step during glycoprotein synthesis. We propose a mechanism for the transport of lipid oligosaccharides through the cell membrane, involving an intermediate stage in which the saccharide moieties are transiently modified with 3-O-methylglucose.
D019325:3-O-Methylglucose; D000241:Adenosine / Q000494:pharmacology; D001426:Bacterial Proteins / Q000032:analysis; D051152:Clusterin; D006023:Glycoproteins / Q000096:biosynthesis*; D006217:Halobacterium / Q000378:metabolism*; D006710:Homocysteine / Q000494:pharmacology; D008745:Methylation; D008757:Methylglucosides / Q000032:analysis / Q000378:metabolism; D018832:Molecular Chaperones; D011104:Polyisoprenyl Phosphate Oligosaccharides / Q000378:metabolism*; D011105:Polyisoprenyl Phosphate Sugars / Q000378:metabolism*
D001426:Bacterial Proteins; D051152:Clusterin; D006023:Glycoproteins; D008757:Methylglucosides; D018832:Molecular Chaperones; D011104:Polyisoprenyl Phosphate Oligosaccharides; D011105:Polyisoprenyl Phosphate Sugars; D006710:Homocysteine; D019325:3-O-Methylglucose; D000241:Adenosine