Preparation and characterisation of fragment glycoasparagines from ovalbumin glycopeptides: Reference compounds for structural and biochemical studies of the oligo-mannose and hybrid types of carbohydrate chains of glycoproteins

Abstract

Glycoasparagines obtained after exhaustive digestion by Pronase of purified ovalbumin were partially degraded by trifluoroacetic acid or subjected to Smith degradation. The partially degraded glycoasparagines thus obtained were first fractionated according to molecular size on Dowex 50W-X2 and then further fractionated by borate chromatography on a column of Sephadex A-25. For a mixture of glycoasparagines of similar molecular size, the latter procedure fractionates according to increasing content of mannosyl cis-2,3-diol. Ten fragment glycoasparagines have been prepared from ovalbumin glycoasparagines, and the structures determined by 1H-n.m.r. spectroscopy and methylation analysis.