Structure of the asn-linked oligosaccharides of apolipophorin III from the insect Locusta migratoria. Carbohydrate-linked 2-aminoethylphosphonate as a constituent of a glycoprotein.
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]
Hård
K
K
Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
Van Doorn
J M
JM
Thomas-Oates
J E
JE
Kamerling
J P
JP
Van der Horst
D J
DJ
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
D001053:Apolipoproteins; D002352:Carrier Proteins; D006023:Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; C042470:apolipophorin III; D001216:Asparagine; D000615:Aminoethylphosphonic Acid