Glycan research on barley, maize, oats, and sorghum grain α-amylases: Comparison with rice α-amylase

• Published in: Archives of biochemistry and biophysics Vol. 278; no. 1; pp. 245 - 250
• Main Authors: Lecommandeur, Didier, Sirou, Yannick, Laurière, Christiane
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.04.1990; Elsevier
• Subjects: AISLAMIENTO (TECNICA); alpha -amylase; alpha-Amylases - isolation & purification; alpha-Amylases - metabolism; AMILASAS; AMYLASE; AMYLASES; AVENA SATIVA; CHROMATOGRAPHIE; CHROMATOGRAPHY; Chromatography, Gel; Concanavalin A; CROMATOGRAFIA; Edible Grain - enzymology; Electrophoresis, Polyacrylamide Gel; Exact sciences and technology; GLICOPROTEINAS; GLYCOPROTEINE; GLYCOPROTEINS; glycosylation; Hordeum - enzymology; HORDEUM VULGARE; hydrophobic interaction chromatography; IMMUNOLOGICAL TECHNIQUES; ISOENZIMAS; ISOENZYME; ISOENZYMES; ISOLATION; ISOLEMENT; Kinetics; Mathematical analysis; Mathematics; Oryza - enzymology; ORYZA SATIVA; Plant Lectins; Plants - enzymology; Potential theory; Sciences and techniques of general use; SORGHUM BICOLOR; Species Specificity; TECHNIQUE IMMUNOLOGIQUE; TECNICAS INMUNOLOGICAS; ZEA MAYS; Zea mays - enzymology; Vegetals; Purification; Hydrophobic chromatography; Enzyme; Secretion; Blotting assay; Immunoblotting assay; Glycosylation; α-Amylase
• ISSN: 0003-9861; 1096-0384
• DOI: 10.1016/0003-9861(90)90254-V

α-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 α-amylase isozymes purified as previously described and using the rice α-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-β- 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 α-amylase constituent was clearly decreased. The same result was obtained after β-elimination in mild conditions. Together these results indicated probable O-linked glycosylation of one maize α-amylase when barley, oats, and sorghum α-amylases did not appear to be glycosylated. Chemical deglycosylation of rice α-amylase resulted in the production of two polypeptides with different molecular weights.