ArticleThe Journal of biological chemistry2022
Exploring the glycosylation of mucins by use of O-glycodomain reporters recombinantly expressed in glycoengineered HEK293 cells.
Article in The Journal of biological chemistry, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 38 citations in OpenAlex.
- Mass Spectrometry-Based Proteomics Methods for Systematic Identification and Quantification of Protein O-Glycosylation in Complex Biological Samples.Journal of the American Society for Mass Spectrometry · 2026Review
- Glycoengineering strategies for constructing defined Mucin O-glycans.Frontiers in molecular biosciences · 2026Review
- Microbial binding module employs sophisticated clustered saccharide patches to selectively adhere to mucins.Nature communications · 2025Article
- Impact of Glycoclustering on Stiffening of MUC5AC Peptides Revealed by High-Efficiency Synthesis.Angewandte Chemie (International ed. in English) · 2025Article
- Glycosylation in T2 high and Th17 Asthma: A Narrative Review.Journal of asthma and allergy · 2025Review
- Adhesion of Pseudomonas aeruginosa to Human Outer Layer Airway Mucins.Methods in molecular biology (Clifton, N.J.) · 2025Article
- UnravelingAnalytical chemistry · 2024Article
- Mammalian cell-based production of glycans, glycopeptides and glycomodules.Nature communications · 2024Article
- Breaking Down the Bottlebrush: Atomically Detailed Structural Dynamics of Mucins.Journal of chemical information and modeling · 2024Article
- Sugar-Coated: Can Multivalent Glycoconjugates Improve upon Nature's Design?Journal of the American Chemical Society · 2024Review
- Goblet cells: guardians of gut immunity and their role in gastrointestinal diseases.eGastroenterology · 2024Article
- Genetics of glycosylation in mammalian development and disease.Nature reviews. Genetics · 2024Review
- B3GNT7 regulates mucin O-glycosylation to alleviate colonic inflammation.BMC gastroenterology · 2024Article
- Binding of Akkermansia muciniphila to mucin is O-glycan specific.Nature communications · 2024Article
- Design of a mucin-selective protease for targeted degradation of cancer-associated mucins.Nature biotechnology · 2024Article
- Mass Spectrometry Strategies forCells · 2024Review
- Mucin adhesion of serial cystic fibrosis airwaysFrontiers in cellular and infection microbiology · 2024Article
- Analysis of complex proteoglycans using serial proteolysis and EThcD provides deep N- and O-glycoproteomic coverage.Analytical and bioanalytical chemistry · 2023Article
- Direct observation of glycans bonded to proteins and lipids at the single-molecule level.Science (New York, N.Y.) · 2023Article
- Molecular Recognition of GalNAc in Mucin-Type O-Glycosylation.Accounts of chemical research · 2023Review
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Authors and funding
11 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Mucins and glycoproteins with mucin-like regions contain densely O-glycosylated domains often found in tandem repeat (TR) sequences. These O-glycodomains have traditionally been difficult to characterize because of their resistance to proteolytic digestion, and knowledge of the precise positions of O-glycans is particularly limited for these regions. Here, we took advantage of a recently developed glycoengineered cell-based platform for the display and production of mucin TR reporters with custom-designed O-glycosylation to characterize O-glycodomains derived from mucins and mucin-like glycoproteins. We combined intact mass and bottom-up site-specific analysis for mapping O-glycosites in the mucins, MUC2, MUC20, MUC21, protein P-selectin-glycoprotein ligand 1, and proteoglycan syndecan-3. We found that all the potential Ser/Thr positions in these O-glycodomains were O-glycosylated when expressed in human embryonic kidney 293 SimpleCells (Tn-glycoform). Interestingly, we found that all potential Ser/Thr O-glycosites in TRs derived from secreted mucins and most glycosites from transmembrane mucins were almost fully occupied, whereas TRs from a subset of transmembrane mucins were less efficiently processed. We further used the mucin TR reporters to characterize cleavage sites of glycoproteases StcE (secreted protease of C1 esterase inhibitor from EHEC) and BT4244, revealing more restricted substrate specificities than previously reported. Finally, we conducted a bottom-up analysis of isolated ovine submaxillary mucin, which supported our findings that mucin TRs in general are efficiently O-glycosylated at all potential glycosites. This study provides insight into O-glycosylation of mucins and mucin-like domains, and the strategies developed open the field for wider analysis of native mucins.
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