Benefits of chemical sugar modifications introduced by click chemistry for glycoproteomic analyses
File(s)
Accepted version
Supporting information
Author(s)
Type
Journal Article
Abstract
Mucin-type O-glycosylation is among the most complex post-translational modifications. Despite mediating many physiological processes, O-glycosylation remains understudied compared to other modifications, simply because the right analytical tools are lacking. In particular, analysis of intact O-glycopeptides by mass spectrometry is challenging for several reasons; O-glycosylation lacks a consensus motif, glycopeptides have low charge density which impairs ETD fragmentation, and the glycan structures modifying the peptides are unpredictable. Recently, we introduced chemically modified monosaccharide analogs that allowed selective tracking and characterization of mucin-type O-glycans after bioorthogonal derivatization with biotin-based enrichment handles. In doing so, we realized that the chemical modifications used in these studies have additional benefits that allow for improved analysis by tandem mass spectrometry. In this work, we built on this discovery by generating a series of new GalNAc analog glycopeptides. We characterized the mass spectrometric signatures of these modified glycopeptides and their MOE signature residues left by bioorthogonal enrichment reagents. Our data indicate that chemical methods for glycopeptide profiling offer opportunities to optimize attributes such as increased charge state, higher charge density, and predictable fragmentation behavior.
Date Issued
2021-09-01
Date Acceptance
2021-04-05
Citation
Journal of the American Society for Mass Spectrometry, 2021, 32 (9), pp.2366-2375
ISSN
1044-0305
Publisher
American Chemical Society
Start Page
2366
End Page
2375
Journal / Book Title
Journal of the American Society for Mass Spectrometry
Volume
32
Issue
9
Copyright Statement
© 2021 American Society for Mass Spectrometry. Published by American Chemical Society. All rights reserved. This document is the Accepted Manuscript version of a Published Work that appeared in final form in J. Am. Soc. Mass Spectrom., after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/jasms.1c00084
Sponsor
Wellcome Trust
Identifier
https://pubs.acs.org/doi/10.1021/jasms.1c00084
Grant Number
218304/Z/19/Z
Subjects
ETD
bioorthogonal
click chemistry
glycopeptide
glycoproteomics
mucin
Analytical Chemistry
0301 Analytical Chemistry
0304 Medicinal and Biomolecular Chemistry
0306 Physical Chemistry (incl. Structural)
Publication Status
Published
Date Publish Online
2021-04-19