title: Salt-adaptation of tobacco BY2 cells induces change in glycoform of N-glycans: enhancement of exo- and endo-glycosidase activities by salt-adaptation.
abstract: In this report, we describe that a salt adaptation of plant cells induces glycoform changes in N-glycoproteins. Intracellular and cell-wall glycopeptides were prepared from glycoproteins expressed in wild-type BY2 cells and salt-adapted cells. N-Glycans were liberated from those glycopeptides by hydrazinolysis, and the released oligosaccharides were N-acetylated and pyridylaminated. The structures of pyridylaminated (PA-) N-glycans were analyzed by a combination of two-dimensional sugar-chain mapping, MS analysis, and exoglycosidase digestion. In both wild-type cells and salt-adapted cells, the plant complex type structure was predominant among N-glycans expressed on glycoproteins, but we found that the Man2Xyl1Fuc1GlcNAc2 structure was significantly expressed on intracellular and cell-wall glycoproteins of the salt-adapted cells. Furthermore, enhancement of the specific activities of alpha-mannosidase and beta-N-acetylglucosaminidase was observed in the salt-adapted BY2 cells, suggesting that the glycoform changes are due to changes in glycosidase activities.
authors.affiliation: Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Japan. yosh8mar@cc.okayama-u.ac.jp
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Kimura
authors.forename: Takao
authors.initials: T
authors.lastname: Watanabe
authors.forename: Mariko
authors.initials: M
authors.lastname: Kimura
authors.forename: Megumi
authors.initials: M
authors.lastname: Maeda
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Murata
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Fujiyama
mesh_terms: D000222:Adaptation, Physiological; D002240:Carbohydrate Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D005959:Glucosidases / Q000378:metabolism*; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D012965:Sodium Chloride / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D014026:Nicotiana / Q000166:cytology / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*
chemical_list: D011134:Polysaccharides; D012965:Sodium Chloride; D005959:Glucosidases