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Organic & Biomolecular Chemistry May 2022Heparan sulfate (HS), a glycosaminoglycan related to heparin, is a linear polysaccharide, consisting of repeating disaccharide units. This compound is involved in...
Heparan sulfate (HS), a glycosaminoglycan related to heparin, is a linear polysaccharide, consisting of repeating disaccharide units. This compound is involved in multiple biological processes such as inflammation, coagulation, angiogenesis and viral infections. Our work focuses on the synthesis of simple HS analogs for the study of structure-activity relationships, with the aim of modulating these biological activities. Thioglycoside analogs, in which the interglycosidic oxygen is replaced by a sulfur atom, are very interesting compounds in terms of therapeutic applications. Indeed, the thioglycosidic bond leads to an improvement of their stability and can allow the inhibition of enzymes involved in physiological and pathological processes. In our previous work, we developed a synthetic sequence which led to a non-sulfated thiodisaccharide analog of HS. In this paper, we report our results of the development of a new synthetic method allowing access to the novel sulfated -disaccharide, as well as to their oxygenated analogues (-disaccharide and sulfated -disaccharide). These 4 compounds were also tested for the inhibition of heparanase, an enzyme involved in biological processes like tumor growth and inflammation. The obtained IC values in the micromolar range showed the impact of the interglycosidic sulfur atom and the 6-sulfate group.
Topics: Disaccharides; Glucuronidase; Heparitin Sulfate; Humans; Inflammation; Sulfur
PubMed: 35388870
DOI: 10.1039/d2ob00250g -
Glycobiology Feb 2021Chondroitin sulfate (CS)and dermatan sulfate (DS) are negatively charged polysaccharides found abundantly in animal tissue and have been extensively described to play...
Chondroitin sulfate (CS)and dermatan sulfate (DS) are negatively charged polysaccharides found abundantly in animal tissue and have been extensively described to play key roles in health and disease. The most common method to analyze their structure is by digestion into disaccharides with bacterial chondroitinases, followed by chromatography and/or mass spectrometry. While studying the structure of oncofetal CS, we noted a large variation in the activity and specificity of commercially available chondroitinases. Here studied the kinetics of the enzymes and used high-performance liquid chromatography-mass spectrometry to determine the di- and oligosaccharide products resulting from the digestion of commercially available bovine CS A, shark CS C and porcine DS, focusing on chondroitinases ABC, AC and B from different vendors. Application of a standardized assay setup demonstrated large variations in the enzyme-specific activity compared to the values provided by vendors, large variation in enzyme specific activity of similar enzymes from different vendors and differences in the extent of cleavage of the substrates and the generated products. The high variability of different chondroitinases highlights the importance of testing enzyme activity and monitoring product formation in assessing the content and composition of chondroitin and DSs in cells and tissues.
Topics: Animals; Carbohydrate Conformation; Cattle; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Dermatan Sulfate; Disaccharides; Substrate Specificity; Swine
PubMed: 32573715
DOI: 10.1093/glycob/cwaa056 -
Methods in Molecular Biology (Clifton,... 2023Chondroitin sulfate proteoglycans (CSPGs) are polyanionic extra/pericellular matrix macromolecules that surround almost all cell types and create microenvironmental...
Chondroitin sulfate proteoglycans (CSPGs) are polyanionic extra/pericellular matrix macromolecules that surround almost all cell types and create microenvironmental niches to support miscellaneous cellular events. In general, the multifunctional properties of CSPGs are attributable to the structural divergence of the CS glycosaminoglycan (GAG) moieties. Because the expression profiles of the GAG chains of CSPGs change with developmental stage, aging, and disease progression, characterization of the GAG chains is essential to understand the functional roles of CSPGs. This chapter describes the basic protocols for GAG moiety-based immunochemical detection of CSPGs in biological samples in conjunction with CS disaccharide composition analysis.
Topics: Chondroitin Sulfate Proteoglycans; Glycosaminoglycans; Disaccharides; Chondroitin Sulfates; Chondroitin
PubMed: 36662459
DOI: 10.1007/978-1-0716-2946-8_2 -
Electrophoresis Jan 2023The feasibility of on-capillary derivatization of saccharides by aromatic amine-based fluorescent labeling agents was tested. To avoid the problematic evolution of...
The feasibility of on-capillary derivatization of saccharides by aromatic amine-based fluorescent labeling agents was tested. To avoid the problematic evolution of gaseous hydrogen cyanide, the Schiff base reduction by sodium cyanoborohydride, as the second step of the standard reductive amination protocol, was omitted. Glucose was used as a model analyte and 7-amino-1,3-naphthalenedisulfonic acid as the labeling agent. Our experiments showed that the direct reaction of the saccharide with the labeling agent in 2.5-M acetic acid yields a labeled product that is sufficiently stable to be separated from the labeling agent in 20-mM phosphate buffer, pH 3.5, and detected using UV detection. The glucose and label zones were introduced separately into the capillary and mixed using a negative voltage. Mixing voltage, its duration, the concentration of acetic acid in the reaction zone, and the waiting time between mixing and separation were optimized. To show the applicability of the procedure to a broader range of analytes, a mixture of different types of saccharides, that is, xylose (pentose), fucose (hexose), glucose (hexose), N-acetylglucosamine (N-acetylaminosaccharide), and lactose (disaccharide), was subjected to derivatization and analysis under the optimal conditions. The linearity and repeatability of the process were evaluated as critical parameters for its analytical applications. Six-point calibration dependences in the 1-50 mM range showed excellent determination coefficients of 0.9992 or higher for all five saccharides tested. The repeatability of the labeled saccharide peak areas was between 2.2% and 4.3%.
Topics: Glucose; Acids; Coloring Agents; Electrophoresis, Capillary; Disaccharides
PubMed: 35699059
DOI: 10.1002/elps.202200136 -
Glycoconjugate Journal Jun 2017Glycosaminoglycans with unique sulfation patterns have been identified in different species of ascidians (sea squirts), a group of marine invertebrates of the Phylum... (Review)
Review
Glycosaminoglycans with unique sulfation patterns have been identified in different species of ascidians (sea squirts), a group of marine invertebrates of the Phylum Chordata, sub-phylum Tunicata (or Urochordata). Oversulfated dermatan sulfate composed of [4-α-L-IdoA-(2-O-SO) → 3-β-D-GalNAc(4-OSO)] repeating disaccharide units is found in the extracellular matrix of several organs, where it seems to interact with collagen fibers. This dermatan sulfate co-localizes with a decorin-like protein, as indicated by immunohistochemical analysis. Low sulfated heparin/heparan sulfate-like glycans composed mainly of [4-α-L-IdoA-(2-OSO) → 4-α-D-GlcN(SO) (6-O-SO)] and [4-α-L-IdoA-(2-O-SO) → 4-α-D-GlcN(SO)] have also been described in ascidians. These heparin-like glycans occur in intracellular granules of oocyte assessory cells, named test cells, in circulating basophil-like cells in the hemolymph, and at the basement membrane of different ascidian organs. In this review, we present an overview of the structure, distribution, extracellular and intracellular localization of the sulfated glycosaminoglycans in different species and tissues of ascidians. Considering the phylogenetic position of the subphylum Tunicata in the phylum Chordata, a careful analysis of these data can reveal important information about how these glycans evolved from invertebrate to vertebrate animals.
Topics: Animal Structures; Animals; Carbohydrate Conformation; Carbohydrate Sequence; Collagen; Decorin; Dermatan Sulfate; Disaccharides; Extracellular Matrix; Hemolymph; Phylogeny; Urochordata
PubMed: 27614617
DOI: 10.1007/s10719-016-9728-5 -
Carbohydrate Polymers Feb 2023Increasing studies focus on chondroitin sulfate (CS) degradation to improve its biological activity. The review mainly introduces the degradation methods of CS and their... (Review)
Review
Increasing studies focus on chondroitin sulfate (CS) degradation to improve its biological activity. The review mainly introduces the degradation methods of CS and their mechanisms. Studies have shown that different degradation methods can lead to different structures of low molecular weight chondroitin sulfate (LMCS). LMCS were prepared through β-elimination reaction, hydrolysis reaction, hydrogen abstraction reaction, and deamination reaction. The degradation of CS is affected by two aspects: the structure of CS (disaccharide composition and molecular weight) and the influence of degradation conditions (temperature, pH, degradation promoters, auxiliary conditions, and time). LMCS with different structures have different biological activities. In addition, degradation could also change CS's metabolism, such as absorption effects and gut microbiota. Thus, choosing the appropriate degradation method for CS development and utilization is very important.
Topics: Chondroitin Sulfates; Disaccharides; Gastrointestinal Microbiome; Hydrogen; Hydrolysis
PubMed: 36446498
DOI: 10.1016/j.carbpol.2022.120361 -
Glycoconjugate Journal Apr 2023Dried leech (Whitmania pigra whitman) has been widely used as a traditional animal-based Chinese medicine. Dried leech extracts have been reported to have various...
Dried leech (Whitmania pigra whitman) has been widely used as a traditional animal-based Chinese medicine. Dried leech extracts have been reported to have various biological activities that are often associated with mammalian glycosaminoglycans. However, their presence and possible structural characteristics within dried leech were previously unknown. In this study, glycosaminoglycans were isolated from dried leech for the first time and their structures were analyzed by the combination of Fourier-transform infrared spectroscopy, liquid chromatography-ion trap/time-of-flight mass spectrometry and polyacrylamide gel electrophoresis. Heparan sulfate and chondroitin sulfate/dermatan sulfate were detected in dried leech with varied disaccharide compositions and possess a heterogeneous structure. Heparan sulfate species possess an equal amount of total 2-O-sulfated, N-sulfated and acetylated disaccharides, while chondroitin sulfate /dermatan sulfate contain high content of 4-O-sulfated disaccharides. Also, the quantitative analysis revealed that the contents of heparan sulfate and chondroitin/dermatan sulfate in dried leech varied significantly, with chondroitin/dermatan sulfate being by far the most abundant. This novel structural information could help clarify the possible involvement of these polysaccharides in the biological activities of the dried leech. Furthermore, leech glycosaminoglycans showed a strong ABTS radical scavenging ability, which suggests the potential of leech polysaccharides for exploitation in the nutraceutical and pharmaceutical industries.
Topics: Animals; Glycosaminoglycans; Chondroitin Sulfates; Dermatan Sulfate; Antioxidants; Heparitin Sulfate; Mammals; Disaccharides
PubMed: 36749437
DOI: 10.1007/s10719-023-10105-y -
Glycoconjugate Journal Oct 2022Galα1 → and GalNAcα1 → are the two essential key sugars in human blood group AB active glycotopes, in which GalNAcα1 → related sequences are located... (Review)
Review
Galα1 → and GalNAcα1 → are the two essential key sugars in human blood group AB active glycotopes, in which GalNAcα1 → related sequences are located at both sides of the nonreducing and the reducing ends of human blood group A active O-glycans. It is also found at the nonreducing ends of GlcNAc N-glycans and glycosphingolipid(GSL) of human blood group A active glycotopes (A) and Forssman antigen (F). When monosaccharides and their α, β anomers are involved in basic units to express the complex size of the combining sites of the GalNAcα1 → specific lectins, they can be divided into a cavity site to accommodate the GalNAcα → key sugar and a subsite with a wide and broad range of recognition area to adopt the rest part of sugar sequences or glycotopes. The function of the subsite is assumed to act as an enhancement factor to increase its affinity power. The following three points are the theme of this mini review: (1) the loci and distribution of the GalNAcα1 → related glycotopes in mammalian glycoconjugates are illustrated and their chemical structures are advanced by the expression of the disaccharide units and code system; (2) the sizes and motifs of GalNAcα1 → specific lectin-glycan interactions are given and (3) the role of the polyvalent blood group A and B glycotopes as blood group AB antigens are proposed. These three highlights should provide an essential background required for the advances in this field.
Topics: Animals; Blood Group Antigens; Disaccharides; Glycoconjugates; Humans; Lectins; Mammals; Polysaccharides
PubMed: 35962217
DOI: 10.1007/s10719-022-10068-6 -
Gut Microbes 2021Human milk glycans present a unique diversity of structures that suggest different mechanisms by which they may affect the infant microbiome development. A humanized...
Human milk glycans present a unique diversity of structures that suggest different mechanisms by which they may affect the infant microbiome development. A humanized mouse model generated by infant fecal transplantation was utilized here to evaluate the impact of fucosyl-α1,3-GlcNAc (3FN), fucosyl-α1,6-GlcNAc, lacto--biose (LNB) and galacto--biose on the fecal microbiota and host-microbiota interactions. 16S rRNA amplicon sequencing showed that certain bacterial genera significantly increased ( and ) or decreased ( and ) in all disaccharide-supplemented groups. Interestingly, cluster analysis differentiates the consumption of fucosyl-oligosaccharides from galactosyl-oligosaccharides, highlighting the disappearance of genus in both fucosyl-oligosaccharides. An increment of the relative abundance of genus was only observed with 3FN. As well, LNB significantly increased the relative abundance of , whereas the absolute levels of this genus, as measured by quantitative real-time PCR, did not significantly increase. OTUs corresponding to the species and were not present in the control after the 3-week intervention, but were shared among the donor and specific disaccharide groups, indicating that their survival is dependent on disaccharide supplementation. The 3FN-feeding group showed increased levels of butyrate and acetate in the colon, and decreased levels of serum HDL-cholesterol. 3FN also down-regulated the pro-inflammatory cytokine TNF-α and up-regulated the anti-inflammatory cytokines IL-10 and IL-13, and the Toll-like receptor 2 in the large intestine tissue. The present study revealed that the four disaccharides show efficacy in producing beneficial compositional shifts of the gut microbiota and in addition, the 3FN demonstrated physiological and immunomodulatory roles.
Topics: Acetates; Adult; Animals; Bacteria; Butyrates; DNA, Bacterial; Disaccharides; Feces; Female; Gastrointestinal Microbiome; Humans; Infant; Infant, Newborn; Male; Mice; Mice, Inbred C57BL; Milk, Human; RNA, Ribosomal, 16S; Young Adult
PubMed: 33938391
DOI: 10.1080/19490976.2021.1914377 -
Glycobiology May 2022Bifidobacterium pseudocatenulatum grows well in the early stages of cultivation in medium containing sucrose (Suc), whereas its growth in medium containing the analogue...
Growth of Bifidobacterium pseudocatenulatum in medium containing N-acetylsucrosamine: enzyme that induces the growth of this bacterium via degradation of this disaccharide.
Bifidobacterium pseudocatenulatum grows well in the early stages of cultivation in medium containing sucrose (Suc), whereas its growth in medium containing the analogue disaccharide N-acetylsucrosamine (SucNAc) tends to exhibit a considerable delay. To elucidate the cause of this phenomenon, we investigated the proliferation pattern of B. pseudocatenulatum in medium containing D-glucose (Glc) and SucNAc and identified the enzyme that degrades this disaccharide. We found that B. pseudocatenulatum initially proliferates by assimilating Glc, with subsequent growth based on SucNAc assimilation depending on production of the β-fructofuranosidase, which can hydrolyze SucNAc, after Glc is completely consumed. Thus, B. pseudocatenulatum exhibited a diauxic growth pattern in medium containing Glc and SucNAc. In contrast, when cultured in medium containing Glc and Suc, B. pseudocatenulatum initially grew by degrading Suc via the phosphorolysis activity of Suc phosphorylase, which did not react to SucNAc. These observations indicate that B. pseudocatenulatum proliferates by assimilating Suc and SucNAc via different pathways. The β-fructofuranosidase of B. pseudocatenulatum exhibited higher hydrolytic activity against several naturally occurring Suc-based tri- or tetrasaccharides than against Suc, suggesting that this enzyme actively catabolizes oligosaccharides other than Suc.
Topics: Bifidobacterium pseudocatenulatum; Disaccharides; Oligosaccharides; Sucrose; beta-Fructofuranosidase
PubMed: 35138388
DOI: 10.1093/glycob/cwac001