Glycobiology最新文献

{"title":"CK2α O-GlcNAcylation affects its conformational stability and modulates its ability to phosphorylate RAD51.","authors":"Nassim El-Kasmi, Alain Defontaine, Damien Marquis, Nizar Ayadi, Mayline Tenou, Cathy Charlier, Fabrice Fleury, Florian Pennarubia","doi":"10.1093/glycob/cwag099","DOIUrl":"https://doi.org/10.1093/glycob/cwag099","url":null,"abstract":"<p><p>Homologous recombination (HR) is a DNA repair pathway which contributes to maintaining genomic integrity. This pathway, which is often dysregulated in cancer treatments resistance, is dependent on RAD51 protein. RAD51 activity is regulated by post-translational modification, particularly its phosphorylation on threonine 13 by casein kinase 2 sub-unit α (CK2α), which facilitates its recruitment to the DNA damage site. CK2α carries an O-GlcNAc, which is known to affect its ability to phosphorylate certain substrates. Recently, several studies have highlighted a modulatory role of O-GlcNAcylation on HR, although no link to RAD51 has been established. Using co-transformation of CK2α with OGT, we obtained an O-GlcNAcylated CK2α protein in bacterio, that we were able to compare to its unmodified counterpart. Structural analyses using Nano DSF and circular dichroism revealed a local stabilizing effect of O-GlcNAcylation on the regions carrying this glycan. Finally, a kinetic comparison of RAD51 phosphorylation by CK2α or CK2α O-GlcNAcylated revealed that O-GlcNAcylation increase CK2α ability to phosphorylate RAD51 at threonine 13. Overall, this study demonstrates for the first time a potential indirect role of O-GlcNAcylation in modulating RAD51 activity.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149216102","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional characterization of a 3-O-sulfate tolerant heparin lyase from Bacteroides eggerthii.","authors":"Taran K Kandola, Courtney J Mycroft-West, Marcelo A Lima, Abigail Turner, Cristina Gardini, Elena Urso, Thomas A Torode, Gavin J Miller, Antonella Bisio, Edwin A Yates, Marco Guerrini, Mark A Skidmore, Liang Wu","doi":"10.1093/glycob/cwag097","DOIUrl":"https://doi.org/10.1093/glycob/cwag097","url":null,"abstract":"<p><p>Heparin is a sulfated glycosaminoglycan, and the most used pharmaceutical anticoagulant worldwide. Heparin is isolated primarily from porcine sources, and possesses a complex variable structure, wherein anticoagulant activity is principally derived from the pentasaccharide GlcNS6S-GlcA-GlcNS3S6S-IdoA2S-GlcNS6S (AGA*IA) motif, containing an infrequent 3-O-sulfated glucosamine that enhances antithrombin stabilization. Pedobacter heparinus heparin lyases are currently widely employed for characterization of heparin and its derivatives, but have limited activity near regions of 3-O-sulfation, hindering analyses of anticoagulation relevant sequences. Here, using a panel of biophysical and structural techniques, we show that Bacteroides eggerthii lyase I liberates ΔUA-GlcNS3S6S disaccharides from the defined pentasaccharide, fondaparinux. This activity provides improved access to 3-O-sulfated heparin standards, with potential to aid heparin quality control.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149180903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multi-Site Quantitative Benchmarking of Neu5Gc-Bearing N-Glycans Using Matrix-Matched LC-MS calibration.","authors":"Nicholas J DeBono, Edward S X Moh, Jessica Poole, Nicolle H Packer, Christopher J Day, Michael P Jennings, Daniel Kolarich, Christopher Ashwood","doi":"10.1093/glycob/cwag088","DOIUrl":"https://doi.org/10.1093/glycob/cwag088","url":null,"abstract":"<p><p>N-glycolylneuraminic acid (Neu5Gc) has been repeatedly associated with human cancer, but reliable detection in mass-spectrometric studies has remained elusive, but its low abundance has led to inconsistent detection in studies of human samples. To address this, matrix-matched calibration curves, which have been pioneered in proteomics and metabolomics for assessing changes in complex mixtures, were measured of released N-glycans at four orders of magnitude dynamic range in defined mixtures, systematically benchmarking Neu5Gc-containing N-glycan detection across multiple LC-MS platforms and sites. Orthogonally, the gold-standard analytical method, consisting of fluorescence detection of labelled monosaccharides separated by LC, was applied to the same samples, yielding absolute concentrations of Neu5Gc. LC-MS demonstrated an extended detection range of three or more orders of magnitude while retaining intact N-glycan measurement, improving assay specificity and enabling detection of the variety of Neu5Gc-bearing N-glycans. By combining orthogonal dimensions of evidence, including chromatographic separation, isotopic distribution matching, and composition-confirming MS/MS, LC-MS confidently resolved Neu5Gc signals from noise, even at low abundance. In comparison, under the DMB-LC-FLR conditions and sample loading evaluated here, dynamic range was limited to two orders of magnitude, insufficient for detection of Neu5Gc in commercially available pooled human sera. These findings strongly support that the DMB-LC-FLR assay's specificity and sensitivity were insufficient for Neu5Gc detection in human samples due to noise overwhelming the Neu5Gc signal. By establishing a reusable benchmarking framework for future glycomic studies, we aim to use LC-MS to improve the measurement of Neu5Gc in clinical samples.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149166668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Milestones in O-GlcNAc Research: Technological Advances that Shaped the Field.","authors":"John A Hanover, Gerald W Hart","doi":"10.1093/glycob/cwag093","DOIUrl":"https://doi.org/10.1093/glycob/cwag093","url":null,"abstract":"<p><p>Since its discovery over forty-years ago, newly developed and improved existing technologies have played a crucial role in advancing O-GlcNAcylation research. We now know that O-GlcNAcylation is central to nutrient/stress regulation of nearly all cellular processes. O-GlcNAcylation was discovered by using labeling by galactosyltransferase and tritiated UDP-GlcNAc combined with product analyses. This enzymatic probe remained the gold standard for early studies in the field. Subsequent development of O-GlcNAc specific antibodies greatly advanced the field by providing non-glycobiologists facile methods for identification of O-GlcNAcylated proteins. Purification and cloning of O-GlcNAc cycling enzymes allowed for development of specific inhibitors and for genetic studies to elucidate the functions of the sugar. Since O-GlcNAc is so labile in the gas phase and is subject to severe ion-suppression in classical collision-induced fragmentation, the development of electron transfer dissociation mass spectrometry was a major advance in O-GlcNAc glycoproteomics, leading to the development of large databases of O-GlcNAcylated proteins. Bioorthogonal chemistry provided valuable new tools to evaluate O-GlcNAc functions in living cells. These and other technological advances have played a major role during the past four decades in establishing O-GlcNAcylation as a major regulator of cellular metabolism.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149136307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemical methods for O-GlcNAc detection and preparation of O-GlcNAc modified proteins.","authors":"Rianna L Haynie-Cion, Michelle Marie B Helmeke, Dongning Liu, Jack T Swanson, Matthew R Pratt","doi":"10.1093/glycob/cwag095","DOIUrl":"10.1093/glycob/cwag095","url":null,"abstract":"<p><p>Chemistry, through the synthesis of a radioactive sugar-donor, played a critical role in the discovery of O-GlcNAc and has been contributing indispensable tools ever since. Here, we cover where chemical methods have contributed to both the discovery and characterization of O-GlcNAc modifications. We will first discuss metabolic labeling and chemoenzymatic modification as two techniques that leverage the power of bioorthogonal chemistry to visualize, enrich, and even quantify O-GlcNAc modifications from living systems. We will then describe how solid-phase peptide synthesis can be combined with protein ligation methods as the only current pathway to homogeneously O-GlcNAc-modified proteins for downstream biological experiments. In each section, we point out the positives and considerations for each approach and provide protocols for normalization of best practices across the field.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149136163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Arterial endothelial expression of α2,8-linked Neu5Gc as a potential target for anti-xeno-antigen inflammation in Cmah-KO mice.","authors":"Motoki Igakura, Yuko Naito-Matsui, Yumina Eguchi, Yuki Yamato, Shuhei Takada, Chihiro Sato, Shogo Oka, Roger A Laine, Hiromu Takematsu","doi":"10.1093/glycob/cwag086","DOIUrl":"https://doi.org/10.1093/glycob/cwag086","url":null,"abstract":"<p><p>N-glycolylneuraminic acid (Neu5Gc), one of the two major sialic acid species in mammals, cannot be biosynthesized in humans because of the inactivation of the responsible enzyme, CMP-Neu5Ac hydroxylase (CMAH). Consequently, Neu5Gc is recognized as an immunologically foreign molecule, resulting in anti-Neu5Gc antibodies production in humans. Meanwhile, dietary Neu5Gc can be metabolically incorporated into glycans and expressed on tissues. Neu5Gc therefore functions as a xeno-autoantigen in humans. The co-existence of Neu5Gc-containing molecules in tissues and circulating anti-Neu5Gc antibodies is thought to drive human-specific chronic inflammation termed xenosialitis and may contribute to vascular disease such as atherosclerosis. Vascular endothelium is a potential major site of Neu5Gc accumulation. Here, we propose the pathological significance of Neu5Gc accumulation likely depends not only on its presence, but also on the glycan structures on which it is displayed. These endothelial Neu5Gc-containing structures remain poorly defined because of the lack of anti-Neu5Gc antibodies to distinguish linkage- and context-specific epitopes. Here, we characterized a vascular Neu5Gc-containing glycan epitope using novel anti-Neu5Gc monoclonal antibodies. Immunohistochemical analyses of mouse tissues showed enriched localization of the epitope for anti-Neu5Gc monoclonal antibody, #23-5, on arterial endothelium. By correlation analysis of glycosyltransferase gene expression and biochemical analyses, the epitope recognized by #23-5 was identified as an α2,8-linked Neu5Gc-containing disialyl structure. In vivo induction of the #23-5 epitope as a xeno-autoantigen was confirmed using Cmah knockout (Cmah-KO) mice after two days of intravenous administration of per-acetylated ManNGc. These findings provide a structural foundation for future studies of Neu5Gc-dependent vascular xenosialitis.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149108531","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Orally administered chondroitin sulfate oligosaccharides attenuate hypertension via multiple protective pathways, including Hmox1-mediated antioxidant responses.","authors":"Hiroko Mizuta, Kosuke Nakamura, Misako Yamaguchi, Masanori Yamaguchi, Nobuyuki Miyamoto","doi":"10.1093/glycob/cwag087","DOIUrl":"https://doi.org/10.1093/glycob/cwag087","url":null,"abstract":"<p><p>Chondroitin sulfate (CS) is a sulfated glycosaminoglycan with various physiological functions. We previously prepared a mixture of chondroitin sulfate oligosaccharides (CSOS) derived from skate cartilage and evaluated its absorption in humans using urinary CS excretion as an indicator. The results suggested that CSOS is absorbed more efficiently than intact high-molecular-weight CS (HMwCS). However, the plasma pharmacokinetics and biological effects of CSOS after administration remained unclear. The present study aimed to clarify these aspects. Pharmacokinetic analysis based on plasma exogenous CS concentrations revealed that CSOS was absorbed, whereas no absorption of HMwCS was observed. Consistent with this finding, a Caco-2 cell assay using CS with different molecular weights showed that lower-molecular-weight CS had greater permeability. To investigate the biological potential of absorbed CSOS, we evaluated its antihypertensive activity in spontaneously hypertensive rats (SHR/Kpo) and explored the underlying mechanisms using microarray analysis. CSOS administration for 12 weeks significantly attenuated hypertension in SHR/Kpo rats. Microarray analysis revealed altered gene expression in analyzed tissues. Among them, we focused on the upregulation of heme oxygenase-1 (Hmox1) in arterial vessels and blood, which has been implicated in oxidative stress-related blood pressure regulation. To evaluate the effect of CSOS on HMOX1 expression, its constituent CS oligomers were examined in EAhy.926 endothelial cells. CS disaccharide (CS2-mer) induced HMOX1 expression, consistent with transcription factor activation. Comparison of CS2-mer analogs indicated that the C6-sulfated disaccharide induced HMOX1 expression without cytotoxicity. These findings suggest that CSOS attenuates hypertension partly through vascular HMOX1 induction, possibly involving the C6-sulfated disaccharide.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149108583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Glyco you should know.","authors":"Manuel M Vicente","doi":"10.1093/glycob/cwag083","DOIUrl":"10.1093/glycob/cwag083","url":null,"abstract":"","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149038221","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The protein sequence modulates terminal GalNAc incorporation during N-glycan biosynthesis.","authors":"Luca Unione, Sandra Delgado, Jesús Jiménez-Barbero, Ana Ardá","doi":"10.1093/glycob/cwag060","DOIUrl":"10.1093/glycob/cwag060","url":null,"abstract":"<p><p>The factors governing protein-specific N-glycosylation remain incompletely understood despite extensive knowledge of the biosynthetic machinery involved in glycan assembly. In particular, the contribution of the protein sequence itself to the final glycosylation pattern has received comparatively little attention. Here, we employed isotope-assisted NMR spectroscopy to investigate the N-glycosylation of the receptor-binding domain of the SARS-CoV-2 spike glycoprotein (RBD-SCoV2) produced in HEK293F cells. Previous analyses revealed the unexpected presence of terminal GalNAc-containing epitopes, including LacdiNAc (LDN), sulfated LacdiNAc (4S-LDN), sialylated LacdiNAc (6Sia-LDN), and fucosylated LacdiNAc (LDNF). Inspired by earlier studies showing that certain β4GalNAc-transferases recognize basic peptide motifs, we generated an RBD variant carrying four substitutions of solvent-exposed basic residues located near the N-glycosylation sites. Comparative NMR analysis demonstrated that the mutant protein exhibits a marked reduction in GalNAc-containing terminal epitopes, accompanied by a corresponding increase in Gal-containing structures. The overall protein fold remained unchanged, indicating that the observed glycosylation differences arise from altered glycan processing rather than major structural perturbations. These findings provide direct evidence that local protein sequence features modulate the incorporation of terminal glycan epitopes during Golgi processing. More broadly, this work highlights the power of quantitative NMR spectroscopy for the structural characterization of glycoproteins and for detecting subtle changes in glycan populations that are directly linked to protein sequence.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13622339/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148669223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Glycan-mediated interactions of infectious hypodermal and hematopoietic necrosis virus (IHHNV) with host tissues in Pacific whiteleg shrimp (Penaeus vannamei).","authors":"Chanyarak Sombutkayasith, Atthaboon Watthammawut, Orawan Thongsum, Jeanne Delafontaine, Supawich Boonkua, Monsicha Somrit, Hiroto Hirayama, Somluk Asuvapongpatana, Clément Delannoy, Coralie Bompard, Shin-Yi Yu, Yannick Rossez, Wattana Weerachatyanukul, Yann Guérardel","doi":"10.1093/glycob/cwag085","DOIUrl":"https://doi.org/10.1093/glycob/cwag085","url":null,"abstract":"<p><p>Infectious hypodermal and hematopoietic necrosis virus (IHHNV) poses a significant threat to global shrimp aquaculture, causing substantial economic losses due to Runt Deformity Syndrome (RDS) in susceptible species, particularly Penaeus vannamei. Despite its widespread impact, the molecular mechanisms underlying IHHNV's tissue tropism and host entry remain poorly understood. This study investigated the role of host glycans in mediating IHHNV attachment and infection, focusing on the interaction between virus-like particles (VLPs) of IHHNV and glycan profiles in P. vannamei tissues. Using recombinant IHHN-VLPs, we demonstrated preferential binding to muscle and gill tissues, with significantly higher affinity compared to the hepatopancreas. Lectin-based assays and mass spectrometry revealed that these tissues were enriched in oligomannosylated N-glycans substituted or not with fucose, terminal N-acetylglucosamine (GlcNAc) and sulfate groups as well as complex O-glycans showing very unusual structural features such as substituted fucose residues and terminal N-acetylgalactosamine residues. Inhibition assays on ELISA, Western blot and histochemistry on shrimp's tissues modified with lectins (ConA, WGA, AAL) and glycosidases (PNGase-F, mannosidase, fucosidase) demonstrated that identified mannosylated and fucosylated glycans were essential for IHHNV recognition. These findings highlight the importance of glycan-mediated interactions in IHHNV infection, suggesting that high-mannose N-glycans and fucosylated O-glycans serve as primary attachment factors. The conservation of these glycan motifs across crustacean species implies their potential as broad-spectrum viral targets. This study not only advances our understanding of IHHNV pathogenesis but also opens avenues for developing glycan-based antiviral strategies, such as competitive inhibitors or targeted glycosidase treatments, to mitigate viral outbreaks in shrimp aquaculture.</p>","PeriodicalId":12766,"journal":{"name":"Glycobiology","volume":" ","pages":""},"PeriodicalIF":3.3,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149074426","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}