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.

J A Van Kuik, R P Sijbesma, J P Kamerling, J F Vliegenthart, E J Wood

Index: Eur. J. Biochem. 169(2) , 399-411, (1987)

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Abstract

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).