Glycoproteins Secreted from Suspension-cultured Tobacco BY2 Cells have Distinct Glycan Structures from Intracellular Glycoproteins(Biochemistry & Molecular Biology)

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Glycan structures of glycoproteins secreted in the spent medium of tobacco BY2 suspension-cultured cells were analyzed. The N-glycans were liberated by hydrazinolysis and the resulting oligosaccharides were labeled with 2-aminopyridine. The pyridylaminated (PA) glycans were purified by reversed-phase and size-fractionation HPLC. The structures of the PA sugar chains were identified by a combination of the two-dimensional PA sugar chain mapping, MS analysis, and exoglycosidase digestion. The ratio (40 : 60) of the amount of glycans with high-mannose-type structure to that with plant-complex-type structure of extracellular glycoproteins is significantly different from that (ratio 10 : 90) previously found in intracellular glycoproteins [Palacpac et al., Biosci Biotechnol. Biochem. 63 (1999) 35-39]. Extracellular glycoproteins have six distinct N-glycans (marked by) from intracellular glycoproteins, and the high-mannose-type structures account for nearly 40% (Man_5GlcNAc_2, 28.8%; Man_6GlcNAc_2, 6.4% ; and Man_7GlcNAc_2, 3.8%), while the plant-complex-type structures account for nearly 60% (GlcNAc_2Man_3 Xyl_1GlcNAc_2, 32.1%; GlcNAc_1Man_3Xyl_1GlcNAc_2 (containing two isomers), 6.2%; GlcNAc_2Man_3 GIcNAc_2, 4.9%; Man_3Xyl_1Fuc_1GlcNAc_2, 8.3% ; and Man_3Xyl_1GlcNAc_2, 3.7%).
社団法人日本農芸化学会の論文
2001-11-23