title: Microbial endo-beta-N-acetylglucosaminidases acting on complex-type sugar chains of glycoproteins.
abstract: The activity and substrate specificity of endo-beta-N-acetylglucosaminidase [glycopeptide-D-mannosyl-N4-(N-acetyl-D-glucosaminyl)2-asparagine 1,4-N-acetyl-beta-glucosamino-hydrolase, EC 3.2.1.96] obtained from Mucor hiemalis (Endo-M) was compared with that of the enzyme obtained from Flavobacterium meningosepticum (Endo-F), which is the only enzyme available that acts on the complex oligosaccharides of asparagine-linked sugar chains in glycoproteins. They showed almost the same activities toward DNS-ovalbumin glycopeptide containing high-mannose and hybrid asparagine-linked oligosaccharides. However, Endo-M showed high activity towards DNS-asialotransferrin and DNS-transferrin glycopeptides, which contain complex biantennary oligosaccharides. Endo-M could weakly act even on DNS-asialofetuin glycopeptide containing complex triantennary oligosaccharides, while Endo-F could not. SDS-denatured asialotransferrin was deglycosylated by both enzymes in the presence of non-ionic detergent (NP-40) and EDTA, and the deglycosylated protein migrated to a lower molecular weight position than asialotransferrin on SDS-PAGE. However, even in the absence of detergent, Endo-M deglycosylated native asialotransferrin and transferrin. Deglycosylation of asialotransferrin was confirmed by means of Con A-Sepharose 4B column chromatography and SDS-PAGE.
authors.affiliation: Kacha College, Kyoto.
authors.forename: S
authors.initials: S
authors.lastname: Kadowaki
authors.forename: K
authors.initials: K
authors.lastname: Yamamoto
authors.forename: M
authors.initials: M
authors.lastname: Fujisaki
authors.forename: T
authors.initials: T
authors.lastname: Tochikura
mesh_terms: D001212:Asialoglycoproteins; D002241:Carbohydrates; D003619:Dansyl Compounds; D060748:Fetuins; D005417:Flavobacterium / Q000201:enzymology; D006023:Glycoproteins; D006868:Hydrolysis; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism*; D009089:Mucor / Q000201:enzymology; D013045:Species Specificity; D013379:Substrate Specificity; D014168:Transferrin / Q000031:analogs & derivatives; D000509:alpha-Fetoproteins
chemical_list: D001212:Asialoglycoproteins; D002241:Carbohydrates; D003619:Dansyl Compounds; D060748:Fetuins; D006023:Glycoproteins; D014168:Transferrin; D000509:alpha-Fetoproteins; C017029:asialofetuin; C017031:asialotransferrins; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase