Open AccessStructure Determination by 360‐MHz 1 H‐NMR Spectroscopy and Methylation Analysis of a Biantennary Glycan of the N ‐Acetyllactosaminic Type Isolated from Rat‐Liver Plasma Membrane
Author(s) -
DEBRAY Henri,
FOURNET Bernard,
MONTREUIL Jean,
DORLAND Lambertus,
VLIEGENTHART Johannes F. C.
Publication year - 1981
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1981.tb06239.x
Subject(s) - chromatography , chemistry , concanavalin a , glycopeptide , elution , sephadex , glycan , size exclusion chromatography , sepharose , nuclear magnetic resonance spectroscopy , fraction (chemistry) , membrane , methylation , biochemistry , glycoprotein , stereochemistry , in vitro , enzyme , antibiotics , gene
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 chromatog raphv 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 α‐glucoside (fraction 3). On the basis of the carbohydrate composition. methylation analysis and 360‐MHz 1 H‐NMR spectroscopy, the following primary structure of a glycan in fraction 2 is proposed: