Open AccessBenefits of Chemical Sugar Modifications Introduced by Click Chemistry for Glycoproteomic Analyses
Author(s) -
Beatriz de la Calle,
Ganka Bineva-Todd,
Andrea Marchesi,
Helen Flynn,
Mattia Ghirardello,
Ömür Y. Tastan,
Chloë Roustan,
Jun Won Choi,
M. Carmen Galan,
Benjamin Schumann,
Stacy A. Malaker
Publication year - 2021
Publication title -
journal of the american society for mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.961
H-Index - 127
eISSN - 1879-1123
pISSN - 1044-0305
DOI - 10.1021/jasms.1c00084
Subject(s) - chemistry , bioorthogonal chemistry , glycosylation , glycan , glycopeptide , isobaric labeling , click chemistry , derivatization , tandem mass spectrometry , mass spectrometry , combinatorial chemistry , proteomics , fragmentation (computing) , biochemistry , isotopic labeling , chromatography , glycoprotein , organic chemistry , protein mass spectrometry , operating system , computer science , gene , antibiotics
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 analogues 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 analogue glycopeptides. We characterized the mass spectrometric signatures of these modified glycopeptides and their signature residues left by bioorthogonal reporter 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.