title: Requirements of cleavage of high mannose oligosaccharides in glycoproteins by peptide N-glycosidase F.
abstract: Peptide N-glycosidase from Flavobacterium meningosepticum cleaves complex as well as neutral glycoproteins (Plummer, T.H., Jr., Elder, J.H., Alexander, S., Phelan, A.W., and Tarentino, A.L. (1984) J. Biol. Chem. 259, 10700-10704). Examples of neutral glycoprotein substrates include ribonuclease B (one high mannose oligosaccharide chain) and yeast external invertase (nine chains/invertase subunit). The rate of deglycosylation by the glycosidase was greatly enhanced if the glycoprotein substrate was denatured prior to enzyme treatment, from a low of 11-fold for external invertase to a high of 844-fold for ribonuclease B. Peptide N-glycosidase F was unable to cleave the asparaginyl-N-acetylglucosamine bond in endo-beta-N-acetylglucosaminidase H-modified external invertase or ribonuclease B, although that in similarly modified glycopeptide substrate was cleaved. Ribonuclease B was digested sequentially with various exoglycosidases to produce an oligosaccharide chain of varied length. Using the resulting forms of ribonuclease B as substrates for peptide N-glycosidase F, the minimum oligosaccharide chain for cleavage was the di-N-acetyl-chitobiosyl core unit.
mesh_terms: D000581:Amidohydrolases / Q000378:metabolism*; D055598:Chemical Phenomena; D002621:Chemistry; D005417:Flavobacterium / Q000201:enzymology; D006023:Glycoproteins / Q000378:metabolism*; D006596:Hexosaminidases / Q000378:metabolism; D006358:Hot Temperature; D008358:Mannose / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011489:Protein Denaturation; D012260:Ribonucleases / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D009844:Oligosaccharides; D012260:Ribonucleases; C029600:ribonuclease B; D006596:Hexosaminidases; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D008358:Mannose