title: Determination of glycosylation sites using a protein sequencer and deglycosylation of native yeast invertase by endo-beta-N-acetylglucosaminidase.
abstract: Endo-beta-N-acetylglucosaminidase from Arthrobacter protophormiae was tested for its capacity to release N-linked sugar chains from native yeast invertase. The enzyme liberated about 80% of the sugar chains from the native invertase. Deglycosylated invertase was digested by chymotrypsin or pepsin, and twelve N-acetylglucosamine-containing glycopeptides were isolated. The amino acid sequences of these glycopeptides were analyzed by a protein sequencer, and the elution position of 4-L-aspartylglycosylamine was directly identified by conventional sequencing. The endo-beta-N-acetylglucosaminidase was found to remove mainly nine sugar chains from native invertase.
authors.affiliation: Department of Bioresource Science, Faculty of Agriculture, Kagawa University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Takegawa
authors.forename: M
authors.initials: M
authors.lastname: Yoshikawa
authors.forename: T
authors.initials: T
authors.lastname: Mishima
authors.forename: M
authors.initials: M
authors.lastname: Nakoshi
authors.forename: S
authors.initials: S
authors.lastname: Iwahara
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006031:Glycosylation; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism*; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D043324:beta-Fructofuranosidase
chemical_list: D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase