Structure determination by 360-MHz 1H-NMR spectroscopy and methylation analysis of a biantennary glycan of the N-acetyllactosaminic type isolated from rat-liver plasma membrane.
Glycopeptides obtained by exhaustive pronase digestion of delipidated rat liver plasmic membranes were purified by gel filtration on Sephadex G-25. These glycopeptides were further fractionated by affinity chromatography on a concanavalin-A--Sepharose 4B column into the following fractions: (a) glycopeptides which did not bind to the column (fraction 1); (b) glycopeptides with weak affinity for concanavalin-A--Sepharose, which could be eluted with buffer only (fraction 2); (c) glycopeptides retained on the column and which could be eluted specifically with buffer containing 0.2 M methyl alpha-glucoside (fraction 3). On the basis of the carbohydrate composition, methylation analysis and 360-MHz 1H-NMR spectroscopy, the following primary structure of a glycan in fraction 2 is proposed: (see formula in text).
D000596:Amino Acids / Q000032:analysis; D000606:Amino Sugars / Q000032:analysis*; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002462:Cell Membrane / Q000378:metabolism; D006020:Glycopeptides / Q000032:analysis; D008099:Liver / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D011134:Polysaccharides / Q000302:isolation & purification; D051381:Rats
D000596:Amino Acids; D000606:Amino Sugars; D006020:Glycopeptides; D011134:Polysaccharides; C000458:N-acetyllactosamine