Demonstration of peptide:N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations.
Endo-beta-N-acetylglucosaminidase F preparations from Flavobacterium meningosepticum have been found to contain peptide:N-glycosidase activity. Only the second activity, designated as peptide:N-glycosidase F, readily cleaves the beta-aspartylglycosylamine linkage of a fetuin triantennary complex glycopeptide, as shown by the isolation of the corresponding carbohydrate-free peptide containing aspartic acid and of an intact oligosaccharide with a di-N-acetylchitobiosyl moiety at the reducing end. Both activities in the mixture will hydrolyze a high mannose octaglycopeptide from ovalbumin, with the type of product formed being influenced by pH. At pH 4.0, only the endo-beta-N-acetylglucosaminidase F activity is functional, releasing octapeptide-GlcNAc and oligosaccharide-GlcNAc. At pH 9.3, the predominant cleavage is by peptide:N-glycosidase F at the glycosylamine bond, releasing octapeptide and oligosaccharide-GlcNAc-GlcNAc. This latter oligosaccharide is then hydrolyzed by endo-beta-N-acetylglucosaminidase F to oligosaccharide-GlcNAc plus GlcNAc.
D000118:Acetylglucosaminidase / Q000302:isolation & purification* / Q000378:metabolism; D000581:Amidohydrolases / Q000302:isolation & purification* / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D005417:Flavobacterium / Q000201:enzymology*; D006020:Glycopeptides / Q000032:analysis; D006595:Hexosamines / Q000032:analysis; D006596:Hexosaminidases / Q000302:isolation & purification*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D010047:Ovalbumin; D010446:Peptide Fragments; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011402:Pronase; D013379:Substrate Specificity; D000509:alpha-Fetoproteins
D000596:Amino Acids; D006020:Glycopeptides; D006595:Hexosamines; D010446:Peptide Fragments; D000509:alpha-Fetoproteins; D010047:Ovalbumin; D006596:Hexosaminidases; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D011402:Pronase; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase