The structure of N-glycans on specific proteins can regulate innate and adaptive immunity via sensing environmental signals. Meanwhile, the structural diversity of N-glycans poses analytical challenges that limit the exploration of specific glycosylation functions. In this work, we used THP-1-derived macrophages as examples to show the vast potential of a N-glycan structural interpretation tool StrucGP in N-glycoproteomic analysis. The intact glycopeptides of macrophages were enriched and analyzed using mass spectrometry (MS)-based glycoproteomic approaches, followed by the large-scale mapping of site-specific glycan structures via StrucGP. Results revealed that bisected GlcNAc, core fucosylated, and sialylated glycans (e.g., HexNAc₄Hex₅Fuc₁Neu5Ac₁, N₄H₅F₁S₁) were increased in M1 and M2 macrophages, especially in the latter. The findings indicated that these structures may be closely related to macrophage polarization. In addition, a high level of glycosylated PD-L1 was observed in M1 macrophages, and the LacNAc moiety was detected at Asn-192 and Asn-200 of PD-L1, and Asn-200 contained Lewis epitopes. The precision structural interpretation of site-specific glycans and subsequent intervention of target glycoproteins and related glycosyltransferases are of great value for the development of new diagnostic and therapeutic approaches for different diseases.
Humans ; B7-H1 Antigen ; Glycosylation ; Polysaccharides/metabolism*

Alzheimer Disease ; metabolism ; Animals ; Antioxidants ; metabolism ; Mice ; Polysaccharides ; pharmacology

Glycosylation is one of the common post-translational modifications of proteins to regulate the ability of tumor invasion, metastasis and tumor heterogeneity by interacting with glycan-binding proteins such as lectins and antibodies. Glycan microarray can be constructed by chemical synthesis, chemical-enzyme synthesis or natural glycan releasing. Glycan microarray is an essential analytical tool to discover the interaction between glycan and its binding proteins. Here we summarize the standard techniques to construct glycan microarray for the application in cancer vaccine, monoclonal antibody and diagnostic markers.
Antibodies, Monoclonal ; Glycosylation ; Lectins/metabolism* ; Microarray Analysis ; Neoplasms ; Polysaccharides

This paper revealed the accumulation regularity of polysaccharides and alcohol-soluble extracts contents of Dendrobium officinale leaves, which have provided basis for the development and utilization of the leaves. The polysaccharides and alcohol-soluble extracts contents of three D. officinale strains leaves collected in different growing periods were determined by phenol-sulfric acid method and hot-dip method respectively. The results showed that the content of polysaccharides in leaves was 4.45% -12.17%, and was about a quarter in stems. The alcohol-soluble extracts content in leaves was 7.45% - 29.34%, and was 1.5 times that of stems. The quality variation of polysaccharides in leaves was closely related to the phenophase. The leaves with lower level of metabolism in three stages: winter, early germination stage and deciduous period, which led to lower content of polysaccharides. The leaves at the vigorous growth stage with higher content of polysaccharides. The alcohol-soluble extracts were closely associated to the formation and germination of buds. The content of alcohol-soluble extracts peaked before sprout, and promoted the growth of new shoots.
Dendrobium ; chemistry ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Plant Leaves ; chemistry ; metabolism ; Polysaccharides ; analysis ; metabolism ; Seasons

OBJECTIVETo establish an ELISA approach to study the interaction of polysaccharides with cytokine in vitro.

METHODSThe heparin BSA complexes (HBC) were synthesized with a chemistry method and separated using a 1 X 90 cm column of Separose 4B. After identification of the complex via SDS-PAGE,the wells of ELISA plates were coated with HBC and the interaction of HBC with interferon-gamma (IFN-gamma) was detected. The effects of heparin, low molecular weight heparin (LMW heparin), chondroitin sulfate (CS), hyaluronic acid (HA) and carrageenans on the binding of HBC to IFN-gamma were tested in this system.

RESULTHuman recombinant IFN-gamma bound to heparin in a concentration dependent manner, the binding of IFN-gamma to HBC was detected at the concentration of 0.25 ng, and saturated at around 2 ng. Free heparin, LMW heparin, CS,HA and carrageenans competed for the binding of IFN-gamma to HBC with significant different ability. The IC(50)concentrations of heparin and LMW heparin were 2.40 microg/ml and 18.60 microg/ml respectively.

CONCLUSIONIFN-gamma is a cytokine with high binding affinity to heparin and carrageenans family but poor to CS-A and CS-C. ELISA is a simple, sensitive approach to detect the interaction of polysaccharides with cytokine in vitro.

Enzyme-Linked Immunosorbent Assay ; methods ; Heparin ; metabolism ; Interferon-gamma ; metabolism ; Polysaccharides ; metabolism ; Serum Albumin, Bovine ; metabolism

OBJECTIVETo study the changes of soluble saccharides and amino acids during the growing period of Coptis chinensis.

METHODAmino acid contents were estimated by Indene tri-ketone colorimetric method; saccharide contents were determined by anthracene ketone method.

RESULT AND CONCLUSIONThe contents of soluble saccharides and amino acids in different organs were low in plants of 1-2 years old. And they were increased in plants of 3-4 years old. At maturation (5 years), the contents of amino acids kept on increasing in rhizome and flower stalk, while decreased in fibre and leaves, and the contents of soluble saccharides showed the downtrend in rhizome, leaves and fibre, except the uptrend in flower stalk.

Amino Acids ; metabolism ; Coptis ; growth & development ; metabolism ; Drugs, Chinese Herbal ; metabolism ; Polysaccharides ; metabolism ; Solubility ; Time Factors

C-type lectins (CTLs) represent a large family of soluble and membrane-bound proteins which bind calcium dependently via carbohydrate recognition domains (CRDs) to glycan residues presented on the surface of a variety of pathogens. The deconvolution of a cell's glycan code by CTLs underpins several important physiological processes in mammals such as pathogen neutralization and opsonization, leukocyte trafficking, and the inflammatory response. However, as our knowledge of CTLs has developed it has become apparent that the role of this innate immune family of proteins can be double-edged, where some pathogens have developed approaches to subvert and exploit CTL interactions to promote infection and sustain the pathological state. Equally, CTL interactions with host glycoproteins can contribute to inflammatory diseases such as arthritis and cancer whereby, in certain contexts, they exacerbate inflammation and drive malignant progression. This review discusses the 'dual agent' roles of some of the major mammalian CTLs in both resolving and promoting infection, inflammation and inflammatory disease and highlights opportunities and emerging approaches for their therapeutic modulation.
Animals ; Humans ; Inflammation/metabolism* ; Lectins, C-Type/metabolism* ; Mammals/metabolism* ; Membrane Proteins ; Polysaccharides/metabolism*

OBJECTIVETo study the polysaccharides contents and monosaccharide compositions in parent root, daughter root and rootlet of Aconitum carmichaeli.

METHODThe conversion coefficient of A. carmichaeli polysaccharide to glucose was obtained by refined polysaccharides, and then the contents of crude polysaccharides in parent root, daughter root and rootlet were determined by sulfuric-phenol spectrometry method; analysis of monosaccharide compositions in polysaccharides from A. carmichaeli was carried out by pre-column derivatization high performance liquid chromatography with 1-phenyl-3-methyl-5-pyrazolone (PMP).

RESULTThe contents of polysaccharides in parent root, daughter root and rootlet were 22.02%, 33.53% and 6.10%, respectively. Parent root, daughter root and rootlet mainly contained glucose, and in addition they contained a small amount of galacturonic acid, galactose and arabinose. Daughter root contained mannose yet, and rootlet still contained mannose, rhamnose and xylose.

CONCLUSIONThe method is simple, rapid, and accurate. The content of polysaccharide in rootlet is lowest, and monosaccharide compositions in rootlet are significantly different from parent root and daughter root.

Aconitum ; metabolism ; Magnetic Resonance Spectroscopy ; Plant Roots ; metabolism ; Polysaccharides ; metabolism ; Reproducibility of Results

To study the effect and mechanism of the light quality acting on Ganoderma lucidum, and provide a theoretical basis for G. lucidum mycelium cultivation, we focused on growth and endogenous IAA metabolism of G. lucidum mycelium under different light-emitting diode (LED) condition. The growth index, endogenous levels of IAA and Enzymes related to IAA metabolism and Polysaccharides content were investigated in different growth periods. Results showed that blue light irradiation was the best from the viewpoint of steady growth and polysaccharides accumulation, red light irradiation improved endogenous IAA level and promoted growth of mycelium in early stage of cultivation, green light irradiation decreased growth rate and fresh weight of mycelium, but increased drying rate. Enzymes related to IAA metabolism also significantly influenced by light quality. The activity of indole acetic acid oxidase (IAAO), peroxidase (POD) and tryptophan synthetase with blue light irradiation were showed high level in early time, but decreased later, and the IAA content was consistently at lower level than that in other treatments, while mycelium irradiated with yellow light showed the highest activity of both IAAO and tryptophan synthetase, and medium level of IAA content. In conclusion, the light quality affects growth and regulation of the level of endogenous IAA of G. lucidum mycelium.
Fungal Polysaccharides ; analysis ; Indoleacetic Acids ; metabolism ; Light ; Peroxidases ; metabolism ; Reishi ; growth & development ; metabolism

Xylitol, a five-carbon sugar alcohol, has many interesting applications in the food, pharmaceutical, and odontological industries, owing to its high sweetening power, its anticariogenic properties, and its insulin-independent metabolism. The bioconversion of detoxified hemicellulosic hydrolysate to xylitol by microorganisms could be a cheaper alternative to the current chemical process, since it is a simple process, with great specificity and low energy requirements. However, the success of fermentations for xylitol production depends on the productivity of the strain and its tolerance to different toxic or inhibitory compounds existing in the hydrolysates. In addition, a number of culture process parameters proved to have significant effects on xylitol production in hemicellulosic hydrolysate media. One of the most important control variables in this bioconversion is the aeration level, which affects the biochemical pathways in the xylose metabolism. The production of biomass is favored by aerobic conditions, while under anaerobic conditions xylose cannot be assimilated by yeast, whereas xylitol is formed in oxygen-limited incubation conditions. An adapted Candida sp. with enhanced resistance to the inhibitors in the hydrolysate can directly ferment the simply detoxified corn cob hemicellulosic hydrolysate to xylitol. In the present study, the combined effects of shaking speed, C/ N ratio, initial pH, and inoculum level on the fermentation of corn cob hemicellulosic hydrolysate to xylitol by an adapted Candida sp. were investigated using an orthogonal experimental design in flask. As a result, the optimum fermentation conditions were as follows: 180 r/min, a C/N ratio of 50, initial pH 5.5, and an inoculum level of 5% (volume ratio). Moreover, the optimum concentration factor of hydrolysate varied between 3.0 and 3.72 was obtained. Based on these results, in order to evaluate the effect of aeration rate on the fermentation of corn cob hemicellulosic hydrolysate to xylitol in fermentor, batch fermentations were carried out in a 3.7 L stirred fermentor using four different aeration strategies, including three kind of two-stage aeration strategies, which provided relatively high aeration rate in the early stage but reduced it in the later stage, and including a one-stage aeration strategy provided a constant aeration rate. With respect to xylitol yield, the results indicated that two-stage aeration strategy was significantly superior to one-stage aeration strategy. The highest xylitol yield (0.75 g/g) was obtained with oxygen supply strategy C (3.75 L/min for first 24 h, then lowered it to 1.25 L/min, 2.5 L fermentation medium was employed). In this process, without extensive detoxification of hydrolysate, an adapted Candida sp. can efficiently ferment the simply treated corn cob hemicellulosic hydrolysate to xylitol under the optimized fermentation conditions. This work should help the development of an efficient process for producing xylitol from corn cob hemicellulosic hydrolysate on a larger scale by bioconversion.
Aerobiosis ; Candida tropicalis ; metabolism ; Fermentation ; Hydrolysis ; Polysaccharides ; metabolism ; Xylitol ; biosynthesis ; Zea mays ; metabolism