title: Degradation of misfolded endoplasmic reticulum glycoproteins in Saccharomyces cerevisiae is determined by a specific oligosaccharide structure.
abstract: In Saccharomyces cerevisiae, transfer of N-linked oligosaccharides is immediately followed by trimming of ER-localized glycosidases. We analyzed the influence of specific oligosaccharide structures for degradation of misfolded carboxypeptidase Y (CPY). By studying the trimming reactions in vivo, we found that removal of the terminal alpha1,2 glucose and the first alpha1,3 glucose by glucosidase I and glucosidase II respectively, occurred rapidly, whereas mannose cleavage by mannosidase I was slow. Transport and maturation of correctly folded CPY was not dependent on oligosaccharide structure. However, degradation of misfolded CPY was dependent on specific trimming steps. Degradation of misfolded CPY with N-linked oligosaccharides containing glucose residues was less efficient compared with misfolded CPY bearing the correctly trimmed Man8GlcNAc2 oligosaccharide. Reduced rate of degradation was mainly observed for misfolded CPY bearing Man6GlcNAc2, Man7GlcNAc2 and Man9GlcNAc2 oligosaccharides, whereas Man8GlcNAc2 and, to a lesser extent, Man5GlcNAc2 oligosaccharides supported degradation. These results suggest a role for the Man8GlcNAc2 oligosaccharide in the degradation process. They may indicate the presence of a Man8GlcNAc2-binding lectin involved in targeting of misfolded glycoproteins to degradation in S. cerevisiae.
authors.affiliation: Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, CH-8091 Zürich, Switzerland.
authors.forename: C A
authors.initials: CA
authors.lastname: Jakob
authors.forename: P
authors.initials: P
authors.lastname: Burda
authors.forename: J
authors.initials: J
authors.lastname: Roth
authors.forename: M
authors.initials: M
authors.lastname: Aebi
mesh_terms: D001483:Base Sequence; D002240:Carbohydrate Sequence; D002268:Carboxypeptidases / Q000378:metabolism*; D043402:Cathepsin A; D004721:Endoplasmic Reticulum / Q000201:enzymology; D005656:Fungal Proteins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D007700:Kinetics; D008361:Mannosidases / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009844:Oligosaccharides / Q000737:chemistry; D017510:Protein Folding; D011499:Protein Processing, Post-Translational / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D000520:alpha-Glucosidases / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D006023:Glycoproteins; D009844:Oligosaccharides; C034251:4-nitrophenyl-alpha-glucosidase; D008361:Mannosidases; C042059:glucosidase I; C046090:mannosyl-oligosaccharide 1,2-alpha-mannosidase; D000520:alpha-Glucosidases; D002268:Carboxypeptidases; D043402:Cathepsin A