The avian influenza A virus receptor SA-α2,3-Gal is expressed in the porcine nasal mucosa sustaining the pig as a mixing vessel for new influenza viruses

Virus Research, Год журнала: 2024, Номер 340, С. 199304 - 199304

Опубликована: Янв. 3, 2024

Influenza A viruses (IAVs) originate from wild birds but have on several occasions jumped host barriers and are now also circulating in humans mammals. The IAV receptors (glycans with galactose linked to a sialic acid (SA) an α2,3 or α2,6 linkage) crucial factors restricting inter-species transmission. In general, avian-origin IAVs show preference for SA-α2,3 (avian receptor), whereas isolated pigs prefer SA-α2,6 (human receptor). N-acetylneuraminic (Neu5Ac) N-glycolylneuraminic (Neu5Gc) the two major SAs. Neu5Ac is expressed all species, Neu5Gc only limited number of domestic species such as horses, not humans. Despite that previous studies shown receptor distribution appears be similar humans, none these investigated expression Neu5Gc-α2,6 situ porcine tissues. Thus, aim this study was elucidate respiratory tract relate viral tropism diverse host-adapted IAVs. (SA-α2,3 SA-α2,6) presence specifically nasal, tracheal, lung tissues by lectin histochemistry. Furthermore, immunohistochemistry performed experimentally infected IAVs, either adapted investigate significance We document first time avian surface nasal mucosa equal Neu5Ac-α2,6 tracheal epithelium alveoli. IAV-infected pigs, we found low amount IAV-positive cells trachea despite high human receptor. Cumulatively, findings suggest optimal replication involves complex interplay between their there might other less clearly defined determine site replication.

Язык: Английский

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Enzymatic modular synthesis of asymmetrically branched human milk oligosaccharides

Carbohydrate Polymers, Год журнала: 2024, Номер 333, С. 121908 - 121908

Опубликована: Фев. 9, 2024

Язык: Английский

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In-situ profiling of glycosylation on single cells with surface plasmon resonance imaging

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Янв. 24, 2025

Cellular glycosylation is crucial for cell recognition, signal transduction, and the development of various diseases, especially in tumor initiation, progression, metastasis. Current profiling methods normally involve laborious sample processing labeling lack in-situ quantitative analysis. Here, we present a direct optical method to investigate quantify glycan expression on single cells based lectin-glycan kinetic quantification with plasmonic imaging. Three unlabeled lectins (WGA, SBA, ConA) are employed as probes bind specific glycans, binding kinetics assessed determine profiles. The result reveals cell-to-cell heterogeneity patterns. To demonstrate capability our method, four distinct lines explored, showing obvious alterations related This approach enables kinetics, providing insights into mechanisms potential applications disease diagnosis treatment. Glycosylation development. authors an optical, single-cell via

Язык: Английский

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EASyMap-Guided Stepwise One-Pot Multienzyme (StOPMe) Synthesis and Multiplex Assays Identify Functional Tetraose-Core-Human Milk Oligosaccharides

JACS Au, Год журнала: 2025, Номер unknown

Опубликована: Янв. 27, 2025

Carbohydrates are biologically and medicinally important molecules that attracting growing attention to their synthesis applications. Unlike the biosynthetic processes for nucleic acids proteins, carbohydrate biosynthesis is not template-driven, more challenging, often leads product variations. In lieu of templates biosynthesis, we describe herein a new concept designing enzyme assembly synthetic maps (EASyMaps) as blueprints guide glycosyltransferase-dependent stepwise one-pot multienzyme (StOPMe) systematically access structurally diverse carbohydrates in target-oriented manner. The strategy demonstrated construction comprehensive library tetraose-core-containing human milk oligosaccharides (HMOs) presenting functional glycan epitopes shared by complex HMOs. tetraose-core-HMOs attractive candidates large-scale production development HMO-based nutraceuticals. To achieve preparative-scale targets containing Neu5Acα2–6GlcNAc component, α2–6-sialyltransferase hST6GALNAC5 successfully expressed E. coli. Neoglycoproteins with controlled valencies prepared immobilized on fluorescent magnetic beads. Multiplex bead assays reveal ligands glycan-binding proteins from plants, influenza viruses, human, bacteria, identifying promising HMO EASyMaps StOPMe systematic manner broadly applicable beyond efficient process suitable can be potentially adapted automation.

Язык: Английский

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Role of Glycans in Equine Endometrial Cell Uptake of Extracellular Vesicles Derived from Amniotic Mesenchymal Stromal Cells

International Journal of Molecular Sciences, Год журнала: 2025, Номер 26(4), С. 1784 - 1784

Опубликована: Фев. 19, 2025

Extracellular vesicles (EVs) are important mediators of cell-cell communication thanks to their ability transfer bioactive cargo, thus regulating a variety physiological contexts. EVs derived from amniotic mesenchymal/stromal cells (eAMC-EVs) internalized by equine endometrial (eECs) with positive effects on regenerative medicine treatments. As the cellular uptake is influenced glycan profile both and target cells, this study focused role surface glycans in eAMC-EVs recipient eECs. Equine ECs were obtained enzymatic digestion uteri healthy mares. AMC-EVs isolated cell cultures according standardized protocol. The pattern was studied using panel lectins combination fucosidase neuraminidase treatment. Both eECs expressed N-linked high mannose glycans, as well fucosylated sialylated glycans. All these involved internalization strongly reduced after cleavage α1,2-linked fucose α2,3/α2,6-linked sialic acids. These results demonstrate that highly relevant molecules

Язык: Английский

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Multiplexed Lectin-PAINT super-resolution microscopy enables cell glycotyping

Communications Biology, Год журнала: 2025, Номер 8(1)

Опубликована: Фев. 20, 2025

Glycosylation profoundly influences cellular function, yet deciphering its intricate patterns remains a formidable challenge. Current techniques often compromise sensitivity, multiplexing, or the ability to capture in-situ cell-to-cell variations. To address these limitations, we introduce 'Lectin-PAINT,' super-resolution imaging method enabling multiplexed live-cell visualization of glycocalyx at single-cell and single-molecule levels. Lectin-PAINT leverages reversible binding lectins specific carbohydrate families perform point accumulation in nanoscale topography (PAINT), identification, mapping, tracking carbohydrates with resolution beyond diffraction limit. Our technique harnesses tailored lectin library, spanning key recognition, offering insights into their abundance, affinity, mobility. Through 8-color imaging, extract more than 350 glycosylation parameters resolution, creating cell's 'glycotype' glycan fingerprint. We showcase power this approach by glycotyping categorizing diverse set cancer cell types, shedding light on heterogeneity variability cancer. In future, research will contribute fundamental understanding changes due disease.

Язык: Английский

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Expedient Assembly of Multiantennary N-Glycans from Common N-Glycan Cores with Orthogonal Protection for the Profiling of Glycan-Binding Proteins

Journal of the American Chemical Society, Год журнала: 2025, Номер unknown

Опубликована: Апрель 7, 2025

Complex-type N-glycans are structurally diverse molecules, responsible for many biological processes, yet the specific sequences of involved in recognition remain largely unknown. Despite recent development efficient chemoenzymatic approaches, it is still lacking a general approach to produce complex-type N-glycans. Here, we designed two common precursors equipped with orthogonal protecting groups antennary differentiation and selective glycan elongation. The N-acetyllactosamine (LacNAc) repeat modules were synthesized separately based on iterative Au(I) promoted glycosylation programmable one-pot strategy incorporated into N-glycan core structure site-specific manner. final removal benzyl was cleanly achieved using pressurized flow chemistry. A total 51 assembled presented as an array study binding specificity toward panel influenza hemagglutinins other lectins. established method allows rapid previously infeasible synthesis asymmetric bi- triantennary N-glycans, especially LacNAc repeats residing at arm, bringing new opportunities carbohydrate-receptor interactions.

Язык: Английский

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Targeting PGM3 abolishes SREBP-1 activation-hexosamine synthesis feedback regulation to effectively suppress brain tumor growth

Science Advances, Год журнала: 2025, Номер 11(16)

Опубликована: Апрель 18, 2025

Elevated hexosamine biosynthesis fuels tumor growth by facilitating protein and lipid glycosylation. But which enzyme in this pathway is better to serve as an antitumor target remains unclear. Here, we revealed that targeting GFAT1, the rate-limiting synthesis, exhibits limited inhibitory effects on glioblastoma (GBM), most lethal brain tumor. This outcome due compensation of NAGK-mediated salvage pathway. Unexpectedly, inhibiting PGM3, controls flux both de novo synthesis pathways, down-regulates expression other enzymes suppresses SREBP-1, a critical lipogenic transcription factor, effectively GBM growth. SREBP-1 transcriptionally up-regulates enzymes, while inhibition these turn activation via reducing N-glycosylation its transporter, SCAP. Our study identified PGM3 promising for treating GBM. Its disrupts activation-hexosamine positive feedback regulation eliminate cells.

Язык: Английский

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Applying transcriptomics to study glycosylation at the cell type level

iScience, Год журнала: 2022, Номер 25(6), С. 104419 - 104419

Опубликована: Май 18, 2022

Язык: Английский

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Glycoproteomics Landscape of Asymptomatic and Symptomatic Human Alzheimer’s Disease Brain

Molecular & Cellular Proteomics, Год журнала: 2022, Номер 21(12), С. 100433 - 100433

Опубликована: Окт. 27, 2022

Molecular changes in the brain of individuals afflicted with Alzheimer's disease (AD) are an intense area study. Little is known about role protein abundance and posttranslational modifications AD progression treatment, particular large-scale intact N-linked glycoproteomics analysis. To elucidate N-glycoproteome landscape, we developed approach based on multi-lectin affinity enrichment, hydrophilic interaction chromatography, LC-MS-based glycoproteomics. We analyzed tissue from 10 persons no cognitive impairment or AD, asymptomatic symptomatic detecting over 300 glycoproteins 1900 glycoforms across samples. The majority have N-glycans that high-mannosidic complex chains fucosylated bisected. Man5 N-glycan was found to occur most frequently at >20% total glycoforms. Unlike glycoproteomes other tissues, sialylation a minor feature N-glycoproteome, occurring <9% among observed AD-associated differences number antennae, frequency fucosylation, bisection, monosaccharides individual glycosylation sites samples our three groups. Further analysis revealed subcellular compartments stage, including lysosome modified paucimannosidic glycans. These results illustrate N-glycoproteomics landscape spectrum clinical pathologic severity will facilitate deeper understanding treatment development.

Язык: Английский

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