title: A rapid mass spectrometric strategy for the characterization of N- and O-glycan chains in the diagnosis of defects in glycan biosynthesis.
abstract: Glycosylation of proteins is a very complex process which involves numerous factors such as enzymes or transporters. A defect in one of these factors in glycan biosynthetic pathways leads to dramatic disorders named congenital disorders of glycosylation (CDG). CDG can affect the biosynthesis of not only protein N-glycans but also O-glycans. The structural analysis of glycans on serum glycoproteins is essential to solving the defect. For this reason, we propose in this paper a strategy for the simultaneous characterization of both N- and O-glycan chains isolated from the serum glycoproteins. The serum (20 microL) is used for the characterization of N-glycans which are released by enzymatic digestion with PNGase F. O-glycans are chemically released by reductive elimination from whole serum glycoproteins using 10 microL of the serum. Using strategies based on mass spectrometric analysis, the structures of N- and O-glycan chains are defined. These strategies were applied on the sera from one patient with CDG type IIa, and one patient with a mild form of congenital disorder of glycosylation type II (CDG-II) that is caused by a deficiency in the Cog1 subunit of the complex.
authors.affiliation: Unité Mixte de Recherche CNRS/USTL 8576, Glycobiologie Structurale et Fonctionnelle, Université des Sciences et Technologies de Lille 1, Villeneuve d'Ascq, France.
authors.forename: Valegh
authors.initials: V
authors.lastname: Faid
authors.forename: Frédéric
authors.initials: F
authors.lastname: Chirat
authors.forename: Nathalie
authors.initials: N
authors.lastname: Seta
authors.forename: François
authors.initials: F
authors.lastname: Foulquier
authors.forename: Willy
authors.initials: W
authors.lastname: Morelle
mesh_terms: D002239:Carbohydrate Metabolism, Inborn Errors / Q000097:blood / Q000175:diagnosis*; D006023:Glycoproteins / Q000097:blood / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D011134:Polysaccharides / Q000032:analysis* / Q000096:biosynthesis / Q000097:blood; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides