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Nature Communications Jun 2024Bacteroidales (syn. Bacteroidetes) are prominent members of the human gastrointestinal ecosystem mainly due to their efficient glycan-degrading machinery, organized into...
Bacteroidales (syn. Bacteroidetes) are prominent members of the human gastrointestinal ecosystem mainly due to their efficient glycan-degrading machinery, organized into gene clusters known as polysaccharide utilization loci (PULs). A single PUL was reported for catabolism of high-mannose (HM) N-glycan glyco-polypeptides in the gut symbiont Bacteroides thetaiotaomicron, encoding a surface endo-β-N-acetylglucosaminidase (ENGase), BT3987. Here, we discover an ENGase from the GH18 family in B. thetaiotaomicron, BT1285, encoded in a distinct PUL with its own repertoire of proteins for catabolism of the same HM N-glycan substrate as that of BT3987. We employ X-ray crystallography, electron microscopy, mass spectrometry-based activity measurements, alanine scanning mutagenesis and a broad range of biophysical methods to comprehensively define the molecular mechanism by which BT1285 recognizes and hydrolyzes HM N-glycans, revealing that the stabilities and activities of BT1285 and BT3987 were optimal in markedly different conditions. BT1285 exhibits significantly higher affinity and faster hydrolysis of poorly accessible HM N-glycans than does BT3987. We also find that two HM-processing endoglycosidases from the human gut-resident Alistipes finegoldii display condition-specific functional properties. Altogether, our data suggest that human gut microbes employ evolutionary strategies to express distinct ENGases in order to optimally metabolize the same N-glycan substrate in the gastroinstestinal tract.
Topics: Polysaccharides; Humans; Gastrointestinal Microbiome; Bacteroides thetaiotaomicron; Bacterial Proteins; Crystallography, X-Ray; Substrate Specificity; Glycoside Hydrolases; Mannose; Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; Multigene Family
PubMed: 38879612
DOI: 10.1038/s41467-024-48802-3 -
Food Science & Nutrition Jun 2024Diguo ( Bur.), an ancient wild fruit, is widely spread in southwest China. However, there is little information on the phenotypic traits, quality characteristics, and...
Diguo ( Bur.), an ancient wild fruit, is widely spread in southwest China. However, there is little information on the phenotypic traits, quality characteristics, and aroma compounds available to diguo fruit. The present study is an investigation into the effects of geographical origin on the phenotypic traits and quality characteristics of wild diguo fruit collected from southwest China. The volatile compounds in the mixed fruit samples were also investigated using gas chromatography-mass spectrometry. Our results indicated that significant variation existed among the sampling materials in all the phenotypic parameters. Fruit fresh weight ranged between 2.06 and 4.59 g. Moreover, significant variation existed among the selected materials in all macronutrients (dry matter, total soluble solids, crude protein, crude fat, and ash) and some nutritional parameters (glutamate, arginine, total soluble solids, maltose, and mannose, etc.). Regardless of their geographical origin, diguo fruit is relatively low in fat and fructose and high in fiber and glutamate. A total of 95 volatile constituents were identified in the frozen diguo fruit. In conclusion, diguo fruit with rich nutritional attributes has a promising future for commercial-scale production. The variability of the observed morphological and nutritional features of diguo fruit provides important characteristics for improving the breeding of diguo as a modern fruit crop.
PubMed: 38873439
DOI: 10.1002/fsn3.4106 -
Cell Surface (Amsterdam, Netherlands) Jun 2024Every fungal cell is encapsulated in a cell wall, essential for cell viability, morphogenesis, and pathogenesis. Most knowledge of the cell wall composition in fungi has...
Every fungal cell is encapsulated in a cell wall, essential for cell viability, morphogenesis, and pathogenesis. Most knowledge of the cell wall composition in fungi has focused on ascomycetes, especially human pathogens, but considerably less is known about early divergent fungal groups, such as species in the Zoopagomycota and Mucoromycota phyla. To shed light on evolutionary changes in the fungal cell wall, we studied the monosaccharide composition of the cell wall of 18 species including early diverging fungi and species in the Basidiomycota and Ascomycota phyla with a focus on those with pathogenic lifestyles and interactions with plants. Our data revealed that chitin is the most characteristic component of the fungal cell wall, and was found to be in a higher proportion in the early divergent groups. The Mucoromycota species possess few glucans, but instead have other monosaccharides such as fucose and glucuronic acid that are almost exclusively found in their cell walls. Additionally, we observed that hexoses (glucose, mannose and galactose) accumulate in much higher proportions in species belonging to Dikarya. Our data demonstrate a clear relationship between phylogenetic position and fungal cell wall carbohydrate composition and lay the foundation for a better understanding of their evolution and their role in plant interactions.
PubMed: 38873189
DOI: 10.1016/j.tcsw.2024.100127 -
Journal of Applied Glycoscience 2024Super Ohtaka, a fermented beverage of plant extracts, is prepared from approximately 50 kinds of fruits and vegetables. Natural fermentation is mainly performed by...
Super Ohtaka, a fermented beverage of plant extracts, is prepared from approximately 50 kinds of fruits and vegetables. Natural fermentation is mainly performed by lactic acid bacteria ( spp.) and yeast ( spp.). Four water-soluble polysaccharide fractions were obtained from Super Ohtaka by dialysis, ion exchange chromatography, and gel filtration chromatography; these fractions were designated as OEP1-1, OEP1-2, OEP2, and OEP3. OEP1-1 is a polysaccharide composed solely of glucose. The other fractions contained polysaccharides composed of glucose, galactose, mannose, and a small amount of arabinose. OEP2 and OEP3 contained phosphorus, which was not detected in OEP1-1 and OEP1-2. Furthermore, the immunomodulatory activity of the polysaccharides was investigated in murine macrophage cell lines. OEP2 and OEP3 significantly induced nitric oxide (NO) secretion by macrophages in a dose-dependent manner (concentration range of 4 to 100 µg/mL). When the concentration of OEP3 was 100 µg/mL, NO production was almost identical to lipopolysaccharide (LPS; 10 ng/mL) used as a positive control. Notably, OEP3 induced NO secretion more strongly than OEP2. This trend was also observed for TNF-α, IL-1β, IL-6, and IL-12 p40 secretion. Overall, our studies on polysaccharides isolated from Super Ohtaka suggest that the fermented beverage stimulates macrophages and activates the immune system.
PubMed: 38863952
DOI: 10.5458/jag.jag.JAG-2023_0012 -
International Journal of General... 2024Mannose receptor C-type 1 (MRC1) is an endocytic lectin receptor primarily expressed in macrophages, dendritic cells, and some endothelial cells. However, the role of...
PURPOSE
Mannose receptor C-type 1 (MRC1) is an endocytic lectin receptor primarily expressed in macrophages, dendritic cells, and some endothelial cells. However, the role of MRC1 in cancers remains unclear.
METHODS
We analyzed MRC1 expression using The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), Cancer Cell Line Encyclopedia (CCLE), and single-cell datasets. We systematically explored the prognostic implications and diagnostic value of MRC1. Immune-related indicators, including immune cells, immune scores, and immune checkpoint molecules, were used to estimate their correlation with MRC1 expression. Finally, we explored its potential ties to immunotherapy success markers such as tumor mutation burden and DNA repair genes.
RESULTS
MRC1 showed both pro- and anti-tumor leanings depending on the cancer types. High levels correlated with poorer outcomes in six cancers but improved prognosis in some cancers like glioblastoma multiforme. This trend extended to the immune arena, where MRC1 intertwined with diverse immune parameters, suggesting its influence on affecting the tumor's immunological landscape. Intriguingly, its expression positively associated with factors favoring immunotherapy efficacy while negatively correlating with some potential barriers. Single-cell analysis pinpointed a specific link between MRC1 and DNA damage/repair pathways in breast cancer.
CONCLUSION
Our study provides a comprehensive landscape of MRC1 levels and diverse regulatory patterns in different cancers, deepening the understanding of MRC1's roles in tumorigenesis and immunity.
PubMed: 38855425
DOI: 10.2147/IJGM.S461144 -
ACS Omega Jun 2024We present a novel colorimetric method inspired by nature's complex mechanisms, capable of selectively determining serotonin with high sensitivity. This method exploits...
We present a novel colorimetric method inspired by nature's complex mechanisms, capable of selectively determining serotonin with high sensitivity. This method exploits the inherent binding affinity of serotonin with sialic acid (SA) molecules anchored to gold nanoparticles (SA-AuNPs). Upon serotonin binding, SA-AuNPs aggregate, and a characteristic red shift in the absorbance of SA-AuNPs accompanied by a dramatic color change (red to blue) occurs, readily observable even without instrumentation. The proposed method effectively eliminates interventions from potential interfering species such as dopamine, epinephrine, l-tyrosine, glucosamine, galactose, mannose, and oxalic acid. The absence of a color change with l-tryptophan, a structurally related precursor of serotonin, further confirms the high selectivity of this approach for serotonin detection. The colorimetric method has a wide linear dynamic range (0.05-1.0 μM), low limit of detection (0.02 μM), and fast response time (5 min). The limit of detection of the method is lower than other colorimetric serotonin sensors reported so far. The possible use of the proposed method in biological sample analysis was evaluated by employing a serotonin recovery assay in processed human plasma. The recoveries ranged from 90.5 to 104.2%, showing promising potential for clinical applications.
PubMed: 38854544
DOI: 10.1021/acsomega.4c01859 -
Molecular & Cellular Proteomics : MCP Jun 2024Protein O-linked mannose (O-Man) glycosylation is an evolutionary conserved post-translational modification (PTM) that fulfills important biological roles during...
Protein O-linked mannose (O-Man) glycosylation is an evolutionary conserved post-translational modification (PTM) that fulfills important biological roles during embryonic development. Three non-redundant enzyme families, POMT1/POMT2, TMTC1-4 and TMEM260, selectively coordinate the initiation of protein O-Man glycosylation on distinct classes of transmembrane proteins, including α-dystroglycan, cadherins and plexin receptors. However, a systematic investigation of their substrate specificities is lacking, in part due to the ubiquitous expression of O-Man glycosyltransferases in cells, which precludes analysis of pathway-specific O-Man glycosylation on a proteome-wide scale. Here, we apply a targeted workflow for membrane glycoproteomics across five human cell lines to extensively map O-Man substrates and genetically deconstruct O-Man initiation by individual and combinatorial knock-out (KO) of O-Man glycosyltransferase genes. We established a human cell library for analysis of substrate specificities of individual O-Man initiation pathways by quantitative glycoproteomics. Our results identify 180 O-Man glycoproteins, demonstrate new protein targets for the POMT1/POMT2 pathway and show that TMTC1-4 and TMEM260 pathways widely target distinct Ig-like protein domains of plasma membrane proteins involved in cell-cell and cell-extracellular matrix interactions. The identification of O-Man on Ig-like folds adds further knowledge on the emerging concept of domain-specific O-Man glycosylation which opens for functional studies of O-Man glycosylated adhesion molecules and receptors.
PubMed: 38851451
DOI: 10.1016/j.mcpro.2024.100796 -
Clinical Proteomics Jun 2024Allergen immunotherapy (AIT) is the only disease-modifying therapy that can achieve immune tolerance in patients through long-term allergen stimulation. Glycans play...
BACKGROUND
Allergen immunotherapy (AIT) is the only disease-modifying therapy that can achieve immune tolerance in patients through long-term allergen stimulation. Glycans play crucial roles in allergic disease, but no information on changes in glycosylation related to an allergic tolerance status has been reported.
METHODS
Fifty-seven patients with house dust mite (HDM) allergies were enrolled. Twenty-eight patients were not treated with AIT, 19 patients had just entered the AIT maintenance treatment phase, and 10 patients had been in the AIT maintenance phase for more than 1 year. Serum protein N-glycans were analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), which included linkage-specific sialylation information.
RESULTS
Eighty-four N-glycans were identified in all three groups. Compared with the patients treated without AIT, the patients treated with AIT for a shorter time showed downregulated expression of high-mannose glycans and upregulated expression of α2,6 sialic acid. The patients treated with AIT in the maintenance phase for over 1 year, which was considered the start of immunological tolerance, showed downregulated expression of biantennary N-glycans and upregulated expression of multibranched and complex N-glycans. Nine N-glycans were changed between allergic and allergic-tolerant patients.
CONCLUSIONS
The glycan form changed from mannose to a more complex type as treatment time increased, and multibranched complex glycans have the potential to be used as a monitoring indicator of immune tolerance. This serum N-glycome analysis provided important information for a deeper understanding of AIT treatment at the molecular level.
PubMed: 38849742
DOI: 10.1186/s12014-024-09491-8 -
BioRxiv : the Preprint Server For... May 2024Coronaviruses recognize a wide array of protein and glycan receptors using the S1 subunit of the spike (S) glycoprotein. The S1 subunit contains two functional domains:...
Coronaviruses recognize a wide array of protein and glycan receptors using the S1 subunit of the spike (S) glycoprotein. The S1 subunit contains two functional domains: the N-terminal (S1-NTD) and C-terminal (S1-CTD). The S1-NTD of SARS-CoV-2, MERS-CoV, and HCoV-HKU1 possess an evolutionarily conserved glycan binding cleft that facilitates weak interactions with sialic acids on cell surfaces. HCoV-HKU1 employs 9--acetylated α2-8-linked disialylated structures for initial binding, followed by TMPRSS2 receptor binding and virus-cell fusion. Here, we demonstrate that HCoV-HKU1 NTD has a broader receptor binding repertoire than previously recognized. We presented HCoV-HKU1 NTD Fc chimeras on a nanoparticle system to mimic the densely decorated surface of HCoV-HKU1. These proteins were expressed by HEK293S GNTI cells, generating species carrying Man-5 structures, often observed near the receptor binding site of CoVs. This multivalent presentation of high-mannose-containing NTD proteins revealed a much broader receptor binding profile compared to its fully glycosylated counterpart. Using glycan microarrays, we observed that 9--acetylated α2-3 linked sialylated LacNAc structures are also bound, comparable to OC43 NTD, suggesting an evolutionarily conserved glycan-binding modality. Further characterization of receptor specificity indicated promiscuous binding towards 9--acetylated sialoglycans, independent of the glycan core (glycolipids, - or -glycans). We demonstrate that HCoV-HKU1 may employ additional sialoglycan receptors to trigger conformational changes in the spike glycoprotein to expose the S1-CTD for proteinaceous receptor binding.
PubMed: 38826377
DOI: 10.1101/2024.05.24.595699 -
Carbohydrate Polymers Sep 2024The influence of locust bean gum (LBG) galactomannans (GMs) molecular weight (Mw) to assemble microparticulate systems was evaluated, and carriers for deep lung delivery...
The influence of locust bean gum (LBG) galactomannans (GMs) molecular weight (Mw) to assemble microparticulate systems was evaluated, and carriers for deep lung delivery were developed. A commercial batch of LBG with a mannose/galactose (M/G) ratio of 2.4 (batch 1) was used to study the influence of different microwave partial acid hydrolysis conditions on carbohydrate composition, glycosidic linkages, and aqueous solutions viscosity. The microwave treatment did not affect the composition, presenting 4-Man (36-42 %), 4,6-Man (27-35 %), and T-Gal (24-25 %) as the main glycosidic linkages. Depolymerization led to a viscosity reduction (≤0.005 Pa·s) with no major impact on polysaccharide debranching. The structural composition of the LBG galactomannans were further elucidated with sequence-specific proteins using carbohydrate microarray technologies. A second batch of LBG (M/G 3.3) was used to study the impact of GMs with different Mw on microparticle assembling, characteristics, and insulin release kinetics. The low-Mw GMs microparticles led to a faster release (20 min) than the higher-Mw (40 min) ones, impacting the release kinetics. All microparticles exhibited a safety profile to cells of the respiratory tract. However, only the higher-Mw GMs allowed the assembly of microparticles with sizes suitable for this type of administration.
Topics: Mannans; Galactose; Molecular Weight; Plant Gums; Humans; Lung; Drug Carriers; Particle Size; Viscosity; Insulin; Drug Liberation; Galactans; Mannose; Animals
PubMed: 38823931
DOI: 10.1016/j.carbpol.2024.122268