title: Structure of the asn-linked oligosaccharides of apolipophorin III from the insect Locusta migratoria. Carbohydrate-linked 2-aminoethylphosphonate as a constituent of a glycoprotein.
abstract: The primary structures of the N-linked carbohydrate chains of apolipophorin III from the insect Locusta migratoria have been determined. The glycoprotein was completely deglycosylated with peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F. Released oligosaccharides were separated from the remaining protein using gel-permeation chromatography on Bio-Gel P-100. Purification of the carbohydrate chains was achieved by a combination of FPLC anion-exchange chromatography on Mono-Q and amine adsorption HPLC on Lichrosorb-NH2. The structures of the carbohydrate chains were deduced with a combination of fast atom bombardment mass spectrometry, 1H- and 31P-NMR spectroscopy, and methylation analysis. The majority of the carbohydrate chains contains 2-aminoethylphosphonate (AEP), which is linked to the 6-position of Man and/or GlcNAc. L. migratoria apolipophorin III is the first example of a glycoprotein containing carbohydrate-linked 2-aminoethylphosphonate. The structures of the major oligosaccharides were established to be the following: [formula: see text]
authors.affiliation: Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Hård
authors.forename: J M
authors.initials: JM
authors.lastname: Van Doorn
authors.forename: J E
authors.initials: JE
authors.lastname: Thomas-Oates
authors.forename: J P
authors.initials: JP
authors.lastname: Kamerling
authors.forename: D J
authors.initials: DJ
authors.lastname: Van der Horst
mesh_terms: D000615:Aminoethylphosphonic Acid / Q000032:analysis*; D000818:Animals; D001053:Apolipoproteins / Q000737:chemistry* / Q000302:isolation & purification; D001216:Asparagine / Q000737:chemistry; D002240:Carbohydrate Sequence; D002352:Carrier Proteins / Q000737:chemistry* / Q000302:isolation & purification; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D006110:Grasshoppers / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D016339:Spectrometry, Mass, Fast Atom Bombardment
chemical_list: D001053:Apolipoproteins; D002352:Carrier Proteins; D006023:Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; C042470:apolipophorin III; D001216:Asparagine; D000615:Aminoethylphosphonic Acid