title: Glycoprotein maturation and the UPR.
abstract: Glycosylation is a complex form of protein modification occurring in the secretory pathway. The addition of N- and O-glycans affects intracellular processes like the folding and trafficking of most glycoproteins. To better understand the impact of glycosylation in protein folding and maturation, parameters like glycosylation site occupancy and oligosaccharide structure must be measured quantitatively. In this chapter, we describe current methods enabling the determination of N-glycosylation by assessment of cellular dolichol phosphate levels, dolichol-linked oligosaccharides, and the occupancy of N-glycosylation sites. We also provide detailed methods for the analysis of O-glycosylation, whose role in intracellular protein maturation is often overlooked.
authors.affiliation: Institute of Physiology, University of Zürich, Winterthurerstrasse, Zürich, Switzerland.
authors.forename: Andreas J
authors.initials: AJ
authors.lastname: Hülsmeier
authors.forename: Michael
authors.initials: M
authors.lastname: Welti
authors.forename: Thierry
authors.initials: T
authors.lastname: Hennet
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004288:Dolichol Phosphates / Q000737:chemistry; D004286:Dolichols / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D009844:Oligosaccharides / Q000032:analysis* / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D056811:Unfolded Protein Response
chemical_list: D004288:Dolichol Phosphates; D004286:Dolichols; D006023:Glycoproteins; D009844:Oligosaccharides