title: Glycan research on barley, maize, oats, and sorghum grain alpha-amylases: comparison with rice alpha-amylase.
abstract: alpha-Amylases from germinated maize, oats, rice, and sorghum were isolated by glycogen precipitation and hydrophobic interaction chromatography. Several methods were used for the detection of glycoproteins, including barley alpha-amylase isozymes purified as previously described and using the rice alpha-amylase as a positive control for glycosylation. Affinoblotting using concanavalin A, immunoblotting using a xylose-specific serum which reacts with complex N-linked glycans, and endo-beta-N-acetylglucosaminidase H treatment of amylases gave negative results for maize, oats, sorghum, and barley. However, after deglycosylation with trifluoromethanesulfonic acid, the molecular weight of one maize alpha-amylase constituent was clearly decreased. The same result was obtained after beta-elimination in mild conditions. Together these results indicated probable O-linked glycosylation of one maize alpha-amylase when barley, oats, and sorghum alpha-amylases did not appear to be glycosylated. Chemical deglycosylation of rice alpha-amylase resulted in the production of two polypeptides with different molecular weights.
authors.affiliation: Laboratoire de Physiologie des Organes Végétaux, CNRS, Meudon, France.
authors.forename: D
authors.initials: D
authors.lastname: Lecommandeur
authors.forename: Y
authors.initials: Y
authors.lastname: Sirou
authors.forename: C
authors.initials: C
authors.lastname: Laurière
mesh_terms: D002850:Chromatography, Gel; D003208:Concanavalin A; D002523:Edible Grain / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D001467:Hordeum / Q000201:enzymology; D007700:Kinetics; D012275:Oryza / Q000201:enzymology; D037121:Plant Lectins; D010944:Plants / Q000201:enzymology*; D013045:Species Specificity; D003313:Zea mays / Q000201:enzymology; D000516:alpha-Amylases / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D037121:Plant Lectins; D003208:Concanavalin A; D000516:alpha-Amylases