title: Analysis of site-specific N-glycan remodeling in the endoplasmic reticulum and the Golgi.
abstract: The hallmark of N-linked protein glycosylation is the generation of diverse glycan structures in the secretory pathway. Dynamic, non-template-driven processes of N-glycan remodeling in the endoplasmic reticulum and the Golgi provide the cellular setting for structural diversity. We applied newly developed mass spectrometry-based analytics to quantify site-specific N-glycan remodeling of the model protein Pdi1p expressed in insect cells. Molecular dynamics simulation, mutational analysis, kinetic studies of in vitro processing events and glycan flux analysis supported the defining role of the protein in N-glycan processing.
authors.affiliation: Institute of Microbiology, Department of Biology.
authors.forename: Ivan
authors.initials: I
authors.lastname: Hang
authors.affiliation: Institute of Microbiology, Department of Biology.
authors.forename: Chia-wei
authors.initials: CW
authors.lastname: Lin
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA.
authors.forename: Oliver C
authors.initials: OC
authors.lastname: Grant
authors.affiliation: Institute of Microbiology, Department of Biology.
authors.forename: Susanna
authors.initials: S
authors.lastname: Fleurkens
authors.affiliation: Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich, Switzerland.
authors.forename: Thomas K
authors.initials: TK
authors.lastname: Villiger
authors.affiliation: Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich, Switzerland.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Soos
authors.affiliation: Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, Swiss Federal Institute of Technology, ETH Zurich, CH-8093 Zurich, Switzerland.
authors.forename: Massimo
authors.initials: M
authors.lastname: Morbidelli
authors.affiliation: Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Woods
authors.affiliation: Institute of Microbiology, Department of Biology markus.aebi@micro.biol.ethz.ch robert.gauss@micro.biol.ethz.ch.
authors.forename: Robert
authors.initials: R
authors.lastname: Gauss
authors.affiliation: Institute of Microbiology, Department of Biology markus.aebi@micro.biol.ethz.ch robert.gauss@micro.biol.ethz.ch.
authors.forename: Markus
authors.initials: M
authors.lastname: Aebi
mesh_terms: D000818:Animals; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: C080576:PDI1 protein, S cerevisiae; D011134:Polysaccharides; D029701:Saccharomyces cerevisiae Proteins; D019704:Protein Disulfide-Isomerases
keywords: Golgi; Golgi glycosylation; endoplasmic reticulum glycosylation; site-specific glycosylation