ArticleAnalytical biochemistry2014
Characterization of intact N- and O-linked glycopeptides using higher energy collisional dissociation.
Article in Analytical biochemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers, 1 of them a synthesis that pooled it.
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Who cites it
38 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Recent Advances in Software Tools for More Generic and Precise Intact Glycopeptide Analysis.Molecular & cellular proteomics : MCP · 2021Pooled it
- Evaluating the Performance of Photon- and Electron-Based Fragmentation Methods in Omnitrap-LCMS Analysis ofAnalytical chemistry · 2026Article
- Collision-Induced Fragmentation of Oligosaccharides: Mechanistic Insights for Mass Spectrometry-Based Glycomics.Angewandte Chemie (International ed. in English) · 2025Review
- Repaglinide restrains HCC development and progression by targeting FOXO3/lumican/p53 axis.Cellular oncology (Dordrecht, Netherlands) · 2024Article
- O-glycosylation of SARS-CoV-2 spike protein by host O-glycosyltransferase strengthens its trimeric structure.Acta biochimica et biophysica Sinica · 2024Article
- Global O-glycoproteome enrichment and analysis enabled by a combinatorial enzymatic workflow.Cell reports methods · 2024Article
- Can We BoostJournal of the American Society for Mass Spectrometry · 2024Article
- Uncovering the fragmentation and separation characteristics of sophorolipid biosurfactants with LC-MS-ESI.Journal of industrial microbiology & biotechnology · 2024Article
- Discovery of Core-Fucosylated Glycopeptides as Diagnostic Biomarkers for Early HCC in Patients with NASH Cirrhosis Using LC-HCD-PRM-MS/MS.ACS omega · 2023Article
- Roles of glycosylation at the cancer cell surface: opportunities for large scale glycoproteomics.Theranostics · 2023Review
- Diversity Matters: Optimal Collision Energies for Tandem Mass Spectrometric Analysis of a Large Set of N-Glycopeptides.Journal of proteome research · 2022Article
- Mass Spectrometry-Based Techniques to Elucidate the Sugar Code.Chemical reviews · 2022Review
- The glycosylation in SARS-CoV-2 and its receptor ACE2.Signal transduction and targeted therapy · 2021Review
- Putting the pieces together: mapping the O-glycoproteome.Current opinion in biotechnology · 2021Review
- What Can N-glycomics and N-glycoproteomics of Cerebrospinal Fluid Tell Us about Alzheimer Disease?Biomolecules · 2021Review
- Towards structure-focused glycoproteomics.Biochemical Society transactions · 2021Review
- Generating orthogonal glycosyltransferase and nucleotide sugar pairs as next-generation glycobiology tools.Current opinion in chemical biology · 2021Review
- Meta-heterogeneity: Evaluating and Describing the Diversity in Glycosylation Between Sites on the Same Glycoprotein.Molecular & cellular proteomics : MCP · 2021Review
- Sequential Analysis of the N/O-Glycosylation of Heavily Glycosylated HIV-1 gp120 Using EThcD-sceHCD-MS/MS.Frontiers in immunology · 2021Article
- A Pragmatic Guide to Enrichment Strategies for Mass Spectrometry-Based Glycoproteomics.Molecular & cellular proteomics : MCP · 2021Review
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Authors and funding
11 authors.
Funding
Abstract
Simultaneous elucidation of the glycan structure and the glycosylation site are needed to reveal the biological function of protein glycosylation. In this study, we employed a recent type of fragmentation termed higher energy collisional dissociation (HCD) to examine fragmentation patterns of intact glycopeptides generated from a mixture of standard glycosylated proteins. The normalized collisional energy (NCE) value for HCD was varied from 30 to 60% to evaluate the optimal conditions for the fragmentation of peptide backbones and glycoconjugates. Our results indicated that HCD with lower NCE values preferentially fragmented the sugar chains attached to the peptides to generate a ladder of neutral loss of monosaccharides, thereby enabling the putative glycan structure characterization. In addition, detection of the oxonium ions enabled unambiguous differentiation of glycopeptides from non-glycopeptides. In contrast, HCD with higher NCE values preferentially fragmented the peptide backbone and, thus, provided information needed for confident peptide identification. We evaluated the HCD approach with alternating NCE parameters for confident characterization of intact N- and O-linked glycopeptides in a single liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. In addition, we applied a novel data analysis pipeline, so-called GlycoFinder, to form a basis for automated data analysis. Overall, 38 unique intact glycopeptides corresponding to eight glycosylation sites (six N-linked and two O-linked sites) were confidently identified from a standard protein mixture. This approach provided concurrent characterization of both the peptide and the glycan, thereby enabling comprehensive structural characterization of glycoproteins in a single LC-MS/MS analysis.
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