title: Porcine pancreatic alpha-amylase shows binding activity toward N-linked oligosaccharides of glycoproteins.
abstract: Porcine pancreatic alpha-amylase was shown by interaction analyses using a resonance mirror detector and alpha-amylase-immobilized Sepharose to bind with glycoproteins possessing N-glycans but not O-linked mucin-type glycans. Direct binding of three types of N-glycans to the alpha-amylase was demonstrated by surface plasmon resonance. Binding with biotin-polymer sugar probes revealed that the alpha-amylase has affinity to alpha-mannose, alpha-N-acetylneuraminic acid, and beta-N-acetyllactosamine, which are components of N-glycans. The binding of glycoproteins or carbohydrates enhanced the enzyme activity, indicating that the recognition site for N-glycans is different from its catalytic site. The binding activity was unique to porcine pancreatic alpha-amylase and was not observed for alpha-amylase from saliva, wheat, and fungus.
authors.affiliation: Course of Advanced Biosciences, Graduate School of Humanities and Sciences, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112-8610, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Matsushita
authors.forename: Mio
authors.initials: M
authors.lastname: Takenaka
authors.forename: Haruko
authors.initials: H
authors.lastname: Ogawa
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry; D002384:Catalysis; D020134:Catalytic Domain; D002417:Cattle; D002845:Chromatography; D005658:Fungi / Q000201:enzymology; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D037102:Lectins; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D010179:Pancreas / Q000201:enzymology*; D037121:Plant Lectins; D011485:Protein Binding; D012463:Saliva / Q000201:enzymology; D020349:Surface Plasmon Resonance; D013552:Swine; D013997:Time Factors; D014908:Triticum / Q000201:enzymology; D000516:alpha-Amylases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D002241:Carbohydrates; D006023:Glycoproteins; D037102:Lectins; D009844:Oligosaccharides; D037121:Plant Lectins; D001710:Biotin; D000516:alpha-Amylases