-
Methods in Molecular Biology (Clifton,... 2022Glycosaminoglycan (GAG) fine structures from the same animal cells and tissues are controlled not only by the biosynthetic and metabolic enzymes but also by other...
Glycosaminoglycan (GAG) fine structures from the same animal cells and tissues are controlled not only by the biosynthetic and metabolic enzymes but also by other environmental factors, such as chemicals, growth factors, nutritional factors, and isolation procedures. To facilitate direct quantitative comparison of disaccharide compositions from different GAG preparations, several stable isotope labeling strategies have been developed. In this report, 1-phenyl-3-methyl-5-pyrazolone (PMP) and deuterated d5-PMP are used for differential disaccharide labeling and profiling of chondroitin sulfate GAG by high performance liquid chromatography (HPLC) coupled with mass spectrometry (MS).
Topics: Animals; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Disaccharides; Glycosaminoglycans; Mass Spectrometry; Polysaccharides
PubMed: 34626374
DOI: 10.1007/978-1-0716-1398-6_10 -
Angewandte Chemie (International Ed. in... Dec 2019The complex sulfation motifs of heparan sulfate glycosaminoglycans (HS GAGs) play critical roles in many important biological processes. However, an understanding of... (Review)
Review
The complex sulfation motifs of heparan sulfate glycosaminoglycans (HS GAGs) play critical roles in many important biological processes. However, an understanding of their specific functions has been hampered by an inability to synthesize large numbers of diverse, yet defined, HS structures. Herein, we describe a new approach to access the four core disaccharides required for HS/heparin oligosaccharide assembly from natural polysaccharides. The use of disaccharides rather than monosaccharides as minimal precursors greatly accelerates the synthesis of HS GAGs, providing key disaccharide and tetrasaccharide intermediates in about half the number of steps compared to traditional strategies. Rapid access to such versatile intermediates will enable the generation of comprehensive libraries of sulfated oligosaccharides for unlocking the "sulfation code" and understanding the roles of specific GAG structures in physiology and disease.
Topics: Disaccharides; Heparitin Sulfate; Humans; Polysaccharides
PubMed: 31553820
DOI: 10.1002/anie.201908805 -
Marine Drugs May 2023The skin is the largest organ of the human body, composed of a diverse range of cell types, non-cellular components, and an extracellular matrix. With aging, molecules...
The skin is the largest organ of the human body, composed of a diverse range of cell types, non-cellular components, and an extracellular matrix. With aging, molecules that are part of the extracellular matrix undergo qualitative and quantitative changes and the effects, such as a loss of skin firmness or wrinkles, can be visible. The changes caused by the aging process do not only affect the surface of the skin, but also extend to skin appendages such as hair follicles. In the present study, the ability of marine-derived saccharides, L-fucose and chondroitin sulphate disaccharide, to support skin and hair health and minimize the effects of intrinsic and extrinsic aging was investigated. The potential of the tested samples to prevent adverse changes in the skin and hair through stimulation of natural processes, cellular proliferation, and production of extracellular matrix components collagen, elastin, or glycosaminoglycans was investigated. The tested compounds, L-fucose and chondroitin sulphate disaccharide, supported skin and hair health, especially in terms of anti-aging effects. The obtained results indicate that both ingredients support and promote the proliferation of dermal fibroblasts and dermal papilla cells, provide cells with a supply of sulphated disaccharide GAG building blocks, increase ECM molecule production (collagen and elastin) by HDFa, and support the growth phase of the hair cycle (anagen).
Topics: Humans; Elastin; Chondroitin Sulfates; Fucose; Cells, Cultured; Skin; Collagen; Fibroblasts; Disaccharides
PubMed: 37367655
DOI: 10.3390/md21060330 -
Proteomics Nov 2019Paucimannosidic N-glycans (PMGs) are special in that they only have the chitobiose core of two β-linked N-acetylglucosamines extended with up to three mannose residues...
Paucimannosidic N-glycans (PMGs) are special in that they only have the chitobiose core of two β-linked N-acetylglucosamines extended with up to three mannose residues and a core fucose. While such short-chain glycans are well described in plants, reports on their expression in humans have been rather scarce. Also, these glycans are often seen as a potential analytical artifact. Recently, it became clear that PMGs are abundantly expressed on various proteins from human neutrophils, opening up a new field of research into the potential biological roles of these modifications. Another niche where PMGs have repeatedly been described are human cancer tissues and cell lines. Chatterjee et al. build on these scattered reports by performing a large-scale pan-cancer investigation showing that PMGs are broadly expressed in different types of tumor tissues as well as related cell lines. Also, the related noncancerous control tissues likewise exhibit PMGs, albeit at lower levels than the corresponding tumors. In conclusion, this study establishes PMGs as a widely occurring modification of human proteins, and further studies are needed to elucidate the biosynthesis, function and tissue- as well as protein-associated expression of PMGs.
Topics: Disaccharides; Glycomics; Glycoproteins; Glycosylation; Humans; Mannose; Polysaccharides; Proteomics; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Tandem Mass Spectrometry
PubMed: 31532879
DOI: 10.1002/pmic.201900244 -
World Journal of Microbiology &... Oct 2023Bifidobacterium is a major probiotic of intestinal gut flora and exerts many physiological activities, and it is widely applied in the fields of food and medicine. As an... (Review)
Review
Bifidobacterium is a major probiotic of intestinal gut flora and exerts many physiological activities, and it is widely applied in the fields of food and medicine. As an important part of Bifidobacterium, glycoside hydrolase plays a role in its physiological activity. With the continuous development and improvement of genetic engineering technology, research on this type of enzyme will play a crucial role in promoting the further development of Bifidobacterium in the field of probiotics. In this review, the preparation methods, enzymatic properties, and functions of glycoside hydrolase extracted from Bifidobacterium are described and summarized. The common method for preparing glycoside hydrolase derived from Bifidobacterium is heterologous expression in Escherichia coli BL21. The optimal pH range for these glycoside hydrolase enzymes is between 4.5 and 7.5; the optimal temperature is between 30 and 50 °C, which is close to the optimal growth condition of Bifidobacterium. Based on substrate specificity, these glycoside hydrolase could hydrolyze synthetic substrates and natural oligosaccharides, including a series of pNP artificial substrates, disaccharide, and trisaccharides, while they have little ability to hydrolyze polysaccharide substrates. This review will be expected to provide a basis for the development of Bifidobacterium as a probiotic element.
Topics: Bifidobacterium; Glycoside Hydrolases; Disaccharides; Oligosaccharides; Substrate Specificity
PubMed: 37843698
DOI: 10.1007/s11274-023-03770-6 -
Journal of Molecular Medicine (Berlin,... Aug 2022Mucopolysaccharidosis type II (MPS II) is a neurometabolic disorder, due to the deficit of the lysosomal hydrolase iduronate 2-sulfatase (IDS). This leads to a severe...
Mucopolysaccharidosis type II (MPS II) is a neurometabolic disorder, due to the deficit of the lysosomal hydrolase iduronate 2-sulfatase (IDS). This leads to a severe clinical condition caused by a multi-organ accumulation of the glycosaminoglycans (GAGs/GAG) heparan- and dermatan-sulfate, whose elevated levels can be detected in body fluids. Since 2006, enzyme replacement therapy (ERT) has been clinically applied, showing efficacy in some peripheral districts. In addition to clinical monitoring, GAG dosage has been commonly used to evaluate ERT efficacy. However, a strict long-term monitoring of GAG content and composition in body fluids has been rarely performed. Here, we report the characterization of plasma and urine GAGs in Ids knock-out (Ids-ko) compared to wild-type (WT) mice, and their changes along a 24-week follow-up, with and without ERT. The concentration of heparan-sulfate (HS), chondroitin-sulfate (CS), and dermatan-sulfate (DS), and of the non-sulfated hyaluronic acid (HA), together with their differentially sulfated species, was quantified by capillary electrophoresis with laser-induced fluorescence. In untreated Ids-ko mice, HS and CS + DS were noticeably increased at all time points, while during ERT follow-up, a substantial decrease was evidenced for HS and, to a minor extent, for CS + DS. Moreover, several structural parameters were altered in untreated ko mice and reduced after ERT, however without reaching physiological values. Among these, disaccharide B and HS 2s disaccharide showed to be the most interesting candidates as biomarkers for MPS II. GAG chemical signature here defined provides potential biomarkers useful for an early diagnosis of MPS II, a more accurate follow-up of ERT, and efficacy evaluations of newly proposed therapies. KEY MESSAGES : Plasmatic and urinary GAGs are useful markers for MPS II early diagnosis and prognosis. CE-LIF allows GAG structural analysis and the quantification of 17 different disaccharides. Most GAG species increase and many structural features are altered in MPS II mouse model. GAG alterations tend to restore to wild-type levels following ERT administration. CS+DS/HS ratio, % 2,4dis CS+DS, and % HS 2s are potential markers for MPS II pathology and ERT efficacy.
Topics: Animals; Biomarkers; Body Fluids; Dermatan Sulfate; Disaccharides; Disease Models, Animal; Enzyme Replacement Therapy; Glycosaminoglycans; Heparitin Sulfate; Mice; Mice, Knockout; Mucopolysaccharidosis II
PubMed: 35816218
DOI: 10.1007/s00109-022-02221-3 -
Critical Care Medicine Nov 2019To determine whether synthetic phosphorylated hexa-acyl disaccharides provide antimicrobial protection in clinically relevant models of bacterial infection. (Comparative Study)
Comparative Study
OBJECTIVES
To determine whether synthetic phosphorylated hexa-acyl disaccharides provide antimicrobial protection in clinically relevant models of bacterial infection.
DESIGN
Laboratory study.
SETTING
University laboratory.
SUBJECTS
BALB/c, C57BL/10J, and C57BL/10ScNJ mice.
INTERVENTIONS
Mice were treated with lactated Ringer's (vehicle) solution, monophosphoryl lipid A, or phosphorylated hexa-acyl disaccharides at 48 and 24 hours prior to intraperitoneal Pseudomonas aeruginosa or IV Staphylococcus aureus infection. Leukocyte recruitment, cytokine production, and bacterial clearance were measured 6 hours after P. aeruginosa infection. In the systemic S. aureus infection model, one group of mice was monitored for 14-day survival and another for S. aureus tissue burden at 3 days postinfection. Duration of action for 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide was determined at 3, 10, and 14 days using a model of intraperitoneal P. aeruginosa infection. Effect of 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide on in vivo leukocyte phagocytosis and respiratory burst was examined. Leukocyte recruitment, cytokine production, and bacterial clearance were measured after P. aeruginosa infection in wild-type and toll-like receptor 4 knockout mice treated with 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide or vehicle to assess receptor specificity.
MEASUREMENTS AND MAIN RESULTS
During intraperitoneal P. aeruginosa infection, phosphorylated hexa-acyl disaccharides significantly attenuated infection-induced hypothermia, augmented leukocyte recruitment and bacterial clearance, and decreased cytokine production. At 3 days post S. aureus infection, bacterial burden in lungs, spleen, and kidneys was significantly decreased in mice treated with monophosphoryl lipid A or phosphorylated hexa-acyl disaccharides, which was associated with improved survival. Leukocyte phagocytosis and respiratory burst functions were enhanced after treatment with monophosphoryl lipid A or phosphorylated hexa-acyl disaccharides. A time course study showed that monophosphoryl lipid A- and 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide-mediated protection against P. aeruginosa lasts for up to 10 days. Partial loss of augmented innate antimicrobial responses was observed in toll-like receptor 4 knockout mice treated with 3-deacyl 6-Acyl phosphorylated hexa-acyl disaccharide.
CONCLUSIONS
Phosphorylated hexa-acyl disaccharides significantly augment resistance against clinically relevant Gram-negative and Gram-positive infections via enhanced leukocyte recruitment, phagocytosis, and respiratory burst functions of innate leukocytes. Improved antimicrobial protection persists for up to 10 days and is partially mediated through toll-like receptor 4.
Topics: Analysis of Variance; Animals; Blotting, Western; Cross Infection; Cytokines; Disaccharides; Disease Models, Animal; Hexosaminidase A; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Peritoneal Cavity; Random Allocation; Staphylococcal Infections; Statistics, Nonparametric; Survival Rate
PubMed: 31567352
DOI: 10.1097/CCM.0000000000003967 -
Carbohydrate Research Dec 2020The indium-mediated allylation followed by ozonolysis has been applied for the elongation of different disaccharides such as cellobiose, lactose and maltose. This...
The indium-mediated allylation followed by ozonolysis has been applied for the elongation of different disaccharides such as cellobiose, lactose and maltose. This reaction sequence and per-O-acetylation produced the expected mixture of α/β-pyranoid as well as α/β-furanoid isomers. The main product in all cases adopted the β-pyranose form and could be isolated and fully characterized with the help of NMR-spin simulations. Thorough investigation of the side products throughout optimization of the conditions for the ozonolysis resulted in the discovery of a novel 12 membered bridged disaccharide.
Topics: Alkenes; Catalysis; Disaccharides; Indium; Isomerism; Ozone
PubMed: 33068775
DOI: 10.1016/j.carres.2020.108170 -
Marine Drugs Oct 2022Fucosylated chondroitin sulfate (FCS) from the sea cucumber (FCS) is the first one that was reported to be branched by disaccharide GalNAc-(α1,2)-Fuc (15%) and...
Fucosylated chondroitin sulfate (FCS) from the sea cucumber (FCS) is the first one that was reported to be branched by disaccharide GalNAc-(α1,2)-Fuc (15%) and sulfated Fuc (85%). Here, four size-homogenous fractions, and seven oligosaccharides, were separated from its β-eliminative depolymerized products. Detailed NMR spectroscopic and MS analyses revealed the oligomers as hexa-, hepta-, octa-, and nonasaccharide, which further confirmed the precise structure of native FCS: it was composed of the CS-E-like backbone with a full content of sulfation at O-4 and O-6 of GalNAc in the disaccharide repeating unit, and the branches consisting of sulfated fucose (Fuc and Fuc) and heterodisaccharide [GalNAc-(α1,2)-Fuc]. Pharmacologically, FCS and its depolymerized derivatives, including fractions and oligosaccharides, showed potent neurite outgrowth-promoting activity in a chain length-dependent manner. A comparison of analyses among oligosaccharides revealed that the sulfate pattern of the Fuc branches, instead of the heterodisaccharide, could affect the promotion intensity. Fuc and the saccharide length endowed the neurite outgrowth stimulation activity most.
Topics: Animals; Sea Cucumbers; Chondroitin Sulfates; Fucose; Oligosaccharides; Disaccharides; Neuronal Outgrowth; Sulfates
PubMed: 36286476
DOI: 10.3390/md20100653 -
Organic & Biomolecular Chemistry Mar 2023Synthetic deoxy-fluoro-carbohydrate derivatives and seleno-sugars are useful tools in protein-carbohydrate interaction studies using nuclear magnetic resonance...
Synthetic deoxy-fluoro-carbohydrate derivatives and seleno-sugars are useful tools in protein-carbohydrate interaction studies using nuclear magnetic resonance spectroscopy because of the presence of the F and Se reporter nuclei. Seven saccharides containing both these atoms have been synthesized, three monosaccharides, methyl 6-deoxy-6-fluoro-1-seleno-β-D-galactopyranoside (1) and methyl 2-deoxy-2-fluoro-1-seleno-α/β-D-galactopyranoside (2α and 2β), and four disaccharides, methyl 4--(β-D-galactopyranosyl)-2-deoxy-2-fluoro-1-seleno-β-D-glucopyranoside (3), methyl 4-Se-(β-D-galactopyranosyl)-2-deoxy-2-fluoro-4-seleno-β-D-glucopyranoside (4), and methyl 4-Se-(2-deoxy-2-fluoro-α/β-D-galactopyranosyl)-4-seleno-β-D-glucopyranoside (5α and 5β), the three latter compounds with an interglycosidic selenium atom. Selenoglycosides 1 and 3 were obtained from the corresponding bromo sugar by treatment with dimethyl selenide and a reducing agent, while compounds 2α/2β, 4, and 5α/5β were synthesized by the coupling of a D-galactosyl selenolate, obtained from the corresponding isoselenouronium salt, with either methyl iodide or a 4--trifluoromethanesulfonyl D-galactosyl moiety. While benzyl ether protecting groups were found to be incompatible with the selenide linkage during deprotection, a change to acetyl esters afforded 4 in a 17% overall yield and over 9 steps from peracetylated D-galactosyl bromide. The synthesis of 5 was performed similarly, but the 2-fluoro substituent led to reduced stereoselectivity in the formation of the isoselenouronium salt (α/β ∼ 1 : 2.3). However, the β-anomer of the uronium salt could be obtained almost pure (∼98%) by precipitation from the reaction mixture. The following displacement reaction occurred without anomerisation, affording, after deacetylation, pure 5β.
Topics: Lactose; Galactose; Disaccharides; Carbohydrate Conformation
PubMed: 36877217
DOI: 10.1039/d2ob02299k