title: alpha-Mannosidase from Trichoderma reesei participates in the postsecretory deglycosylation of glycoproteins.
abstract: The 160 kDa alpha-mannosidase (E.C. 3.2.1.24) isolated from culture filtrate of Trichoderma reesei has wide aglycon specificity but cleaves the alpha1 --> 2 and alpha1 --> 3 mannosidic bonds with higher rate than alpha1 --> 6 bond and slowly hydrolyses yeast mannan and 1,6-alpha-mannan. The specific activity of the enzyme and rate constant in the reaction with p-nitrophenyl-alpha-D-mannopyranoside were 0.15 U/mg and 1.62 x 10(-4) microM/min/microg, respectively, at optimal pH 6.5. We have found that in vitro enzyme is able to cleave off 30% of total alpha-mannopyranosyl residues from N- and O-linked glycans of secreted glycoproteins. The activity of the alpha-mannosidase toward glycoproteins in vivo was studied comparing the structures of O- and N-linked glycans of glycoproteins isolated from the cultures growing with and without 1-deoxymannojirimycin, an inhibitor of alpha-mannosidases. Difference in structures of these glycans may be explained by postsecretory deglycosylation catalysed by the alpha-mannosidase.
authors.affiliation: Petersburg Nuclear Physics Institute, Gatchina, St. Petersburg, 188350, Russia.
authors.forename: E V
authors.initials: EV
authors.lastname: Eneyskaya
authors.forename: A A
authors.initials: AA
authors.lastname: Kulminskaya
authors.forename: A N
authors.initials: AN
authors.lastname: Savel'ev
authors.forename: K A
authors.initials: KA
authors.lastname: Shabalin
authors.forename: A M
authors.initials: AM
authors.lastname: Golubev
authors.forename: K N
authors.initials: KN
authors.lastname: Neustroev
mesh_terms: D017485:1-Deoxynojirimycin / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005656:Fungal Proteins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D007700:Kinetics; D008351:Mannans / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D008361:Mannosidases / Q000378:metabolism*; D013379:Substrate Specificity; D014242:Trichoderma / Q000201:enzymology*; D043323:alpha-Mannosidase
chemical_list: D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D006023:Glycoproteins; D008351:Mannans; D017485:1-Deoxynojirimycin; D008361:Mannosidases; D043323:alpha-Mannosidase; D008358:Mannose