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Article Dans Une Revue Rapid Communications in Mass Spectrometry Année : 2015

Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid

Résumé

Rationale Glycosylation is one of the most complex types of post‐translational modifications of proteins. The alteration of glycans bound to proteins from cerebrospinal fluid (CSF) in relation to disorders of the central nervous system is a highly relevant subject, but only few studies have focused on the glycosylation of CSF proteins. Methods Reproducible profiles of CSF N‐glycans were first obtained by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry after permethylation. Tryptic glycopeptides from CSF proteins were also enriched by hydrophilic interaction, and the resulting extracts divided into two equal aliquots. A first aliquot was enzymatically deglycosylated and analyzed by nano‐liquid chromatography/tandem mass spectrometry while the second one, containing intact enriched glycopeptides, was directly analyzed. Site‐specific data were obtained by combining the data from these three experiments. Results We describe the development of a versatile approach for obtaining site‐specific information on the N‐glycosylation of CSF glycoproteins. Under these conditions, 124 N‐glycopeptides representing 55 N‐glycosites from 36 glycoproteins were tentatively identified. Special emphasis was placed on the analysis of glycoproteins/glycopeptides bearing 'brain‐type' N‐glycans, representing potential biologically relevant structures in the field of neurodegenerative disorders. Using our workflow, only a few proteins were shown to carry such particular glycan motifs. Conclusions We developed an approach combining N‐glycomics and N‐glycoproteomics and underline its usefulness to study the site‐specific glycosylation of major human CSF proteins. The final rather long‐term objective is to combine these data with those from other omics approaches to delve deeper into the understanding of particular neurological disorders. Copyright © 2015 John Wiley & Sons, Ltd.

Dates et versions

hal-04403907 , version 1 (18-01-2024)

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Citer

Arnaud Goyallon, Sophie Cholet, Manuel Chapelle, Christophe Junot, François Fenaille. Evaluation of a combined glycomics and glycoproteomics approach for studying the major glycoproteins present in biofluids: Application to cerebrospinal fluid. Rapid Communications in Mass Spectrometry, 2015, 29 (6), pp.461-473. ⟨10.1002/rcm.7125⟩. ⟨hal-04403907⟩
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