Presence of asparagine-linked N-acetylglucosamine and chitobiose in Pyrus pyrifolia S-RNases associated with gametophytic self-incompatibility

S-RNases encoded by the S-locus of rosaceous and solanaceous plants discriminate between the S-alleles of pollen in gametophytic self-incompatibility reactions, but it is not clear how. We report the structures of N-glycans attached to each of the N-glycosylation sites of seven S-RNases in Pyr us pyrifolia of the Rosaceae. The structures were identified by chromatographic analysis of pyridyiaminnted sugar chains prepared from S-4-RNase and by liquid chromatography/electrospray ionization-mass spectrometric analysis of the protease digests of reduced and S-carboxymethylated S-RNases. S-4-RNase carries various types of sugar chains, including plant-specific ones with beta 1-->2-linked xylose and alpha 1-->3-linked fucose residues. More than 70% of the total N-glycans of S-4-RNase are, however, an N-acetylglucosamine or a chitobiose (GlcNAc beta 1-->4GlcNAc), which has not been found naturally. The N-acetylglucosamine and chitobiose are mainly present at the N-glycosylation sites within the putative recognition sites of the S-RNase, suggesting that these sugar chains may interact with pollen S-product(s).