Primary structure determination of seven novel N-linked carbohydrate chains derived from hemocyanin of Lymnaea stagnalis. 3-O-methyl-D-galactose and N-acetyl-D-galactosamine as constituents of xylose-containing N-linked oligosaccharides in an animal glycoprotein.
Hemocyanin from the freshwater snail Lymnaea stagnalis is a high-molecular-mass copper-containing glyco-protein which functions as oxygen carrier in the hemolymph. To release the carbohydrate chains, the protein was digested by pronase followed by hydrazinolysis and reduction. The oligosaccharide-alditols were purified by gel permeation chromatography on Bio-Gel P-4, followed by HPLC on a Lichrosorb-NH2 column. Using 500-MHz 1H-NMR spectroscopy, in conjunction with sugar, methylation and deamination analysis, the following seven novel primary oligosaccharide structures could be unravelled. (Formula: see text).
Department of Bio-Organic Chemistry, Transitorium III, Utrecht University, The Netherlands.
J A
JA
Van Kuik
R P
RP
Sijbesma
J P
JP
Kamerling
J F
JF
Vliegenthart
E J
EJ
Wood
D000116:Acetylgalactosamine / Q000032:analysis*; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D005688:Galactosamine / Q000031:analogs & derivatives*; D006433:Hemocyanins; D007202:Indicators and Reagents; D008195:Lymnaea; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008756:Methylgalactosides / Q000032:analysis*; D008759:Methylglycosides / Q000032:analysis*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification; D014994:Xylose / Q000032:analysis*
D007202:Indicators and Reagents; D008756:Methylgalactosides; D008759:Methylglycosides; D009844:Oligosaccharides; C031552:methyl beta-galactoside; D005688:Galactosamine; D006433:Hemocyanins; D014994:Xylose; D000116:Acetylgalactosamine