Carbohydrate Structures of the Glycoprotein Allergen Cry j I from Japanese Cedar (Cryptomeria japonica) Pollen

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Abstract
The glycoprotein allergen Cry j I from Japanese cedar (Cryptomeria japonica) pollen was treated with pepsin and glycopeptidase A to release asparagine-linked oligosaccharides. The reducing ends of the oligosaccharides were aminated with the fluorescent reagent 2-aminopyridine. The oligosaccharide derivatives were purified by gel permeation chro matography and reversed-phase HPLC. Their structures were determined by sequential exoglycosidase digestion and 500 MHz H-NMR spectroscopy. Four oligosaccharide struc tures, A, B, C, and D, were identified as the xylose-containing complex-type. They were present at a molar ratio of 8: 1 : 6: 1. By amino acid sequence analyses of the tryptic peptides, Asn-170 and Asn-333 of Cry j I were found to carry asparagine-linked oligosac charides.

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