Ergosteryl-{beta}-glucosidase (Egh1) involved in sterylglucoside catabolism and vacuole formation in Saccharomyces cerevisiae
Sterylglucosides (SGs) are composed of a glucose and sterol derivatives, and are distributed in fungi, plants and mammals. We recently identified EGCrP1 and EGCrP2 (endoglycoceramidase-related proteins 1 and 2) as a β-glucocerebrosidase and steryl-β-glucosidase, respectively, in Cryptococcus neoformans. We herein describe an EGCrP2 homologue (Egh1; ORF name, Yir007w) involved in SG catabolism in Saccharomyces cerevisiae. The purified recombinant Egh1 hydrolyzed various β-glucosides including ergosteryl β-glucoside (EG), cholesteryl β-glucoside, sitosteryl β-glucoside, para-nitrophenyl β-g...
Source: Glycobiology - August 27, 2015 Category: Biology Authors: Watanabe, T., Tani, M., Ishibashi, Y., Endo, I., Okino, N., Ito, M. Tags: Glycan Metabolism Source Type: research
In-depth N-glycome profiling of paired colorectal cancer and non-tumorigenic tissues reveals cancer-, stage- and EGFR-specific protein N-glycosylation
Glycomics may assist in uncovering the structure–function relationships of protein glycosylation and identify glycoprotein markers in colorectal cancer (CRC) research. Herein, we performed label-free quantitative glycomics on a carbon-liquid chromatography–tandem mass spectrometry-based analytical platform to accurately profile the N-glycosylation changes associated with CRC malignancy. N-Glycome profiling was performed on isolated membrane proteomes of paired tumorigenic and adjacent non-tumorigenic colon tissues from a cohort of five males (62.6 ± 13.1 y.o.) suffering from colorectal adenocarcinoma. Th...
Source: Glycobiology - August 27, 2015 Category: Biology Authors: Sethi, M. K., Kim, H., Park, C. K., Baker, M. S., Paik, Y.-K., Packer, N. H., Hancock, W. S., Fanayan, S., Thaysen-Andersen, M. Tags: Cancer Biology Source Type: research
EndoS and EndoS2 hydrolyze Fc-glycans on therapeutic antibodies with different glycoform selectivity and can be used for rapid quantification of high-mannose glycans
Enzymes that affect glycoproteins of the human immune system, and thereby modulate defense responses, are abundant among bacterial pathogens. Two endoglycosidases from the human pathogen Streptococcus pyogenes, EndoS and EndoS2, have recently been shown to hydrolyze N-linked glycans of human immunoglobulin G. However, detailed characterization and comparison of the hydrolyzing activities have not been performed. In the present study, we set out to characterize the enzymes by comparing the activities of EndoS and EndoS2 on a selection of therapeutic monoclonal antibodies (mAbs), cetuximab, adalimumab, panitumumab and denosu...
Source: Glycobiology - August 27, 2015 Category: Biology Authors: Sjogren, J., Cosgrave, E. F. J., Allhorn, M., Nordgren, M., Bjork, S., Olsson, F., Fredriksson, S., Collin, M. Tags: Analytical Glycobiology Source Type: research
Distinct structures of the {alpha}-fucose branches in fucosylated chondroitin sulfates do not affect their anticoagulant activity
Fucosylated chondroitin sulfate (FCS) is a glycosaminoglycan found in sea cucumbers. It has a backbone like that of mammalian chondroitin sulfate (4-β-d-GlcA-1->3-β-d-GalNAc-1)n but substituted at the 3rd position of the β-d-glururonic acid residues with α-fucose branches. The structure of these branches varies among FCSs extracted from different species of sea cucumbers, as revealed by solution NMR spectroscopy. Some species (Isostichopus badionotus and Patalus mollis) contain branches formed by single α-fucose residues but with variable sulfation patterns (2,4-, 3,4- and 4-sulfation). FCS fr...
Source: Glycobiology - August 27, 2015 Category: Biology Authors: Santos, G. R. C., Glauser, B. F., Parreiras, L. A., Vilanova, E., Mourao, P. A. S. Tags: Analytical Glycobiology Source Type: research
The multiple roles of epidermal growth factor repeat O-glycans in animal development
The epidermal growth factor (EGF)-like repeat is a common, evolutionarily conserved motif found in secreted proteins and the extracellular domain of transmembrane proteins. EGF repeats harbor six cysteine residues which form three disulfide bonds and help generate the three-dimensional structure of the EGF repeat. A subset of EGF repeats harbor consensus sequences for the addition of one or more specific O-glycans, which are initiated by O-glucose, O-fucose or O-N-acetylglucosamine. These glycans are relatively rare compared to mucin-type O-glycans. However, genetic experiments in model organisms and cell-based assays indi...
Source: Glycobiology - August 27, 2015 Category: Biology Authors: Haltom, A. R., Jafar-Nejad, H. Tags: REVIEW Source Type: research
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Source: Glycobiology - August 27, 2015 Category: Biology Tags: Cover/Standing Material Source Type: research
Crystal structure of a symbiosis-related lectin from octocoral
D-Galactose-binding lectin from the octocoral, Sinularia lochmodes (SLL-2), distributes densely on the cell surface of microalgae, Symbiodinium sp., an endosymbiotic dinoflagellate of the coral, and is also shown to be a chemical cue that transforms dinoflagellate into a non-motile (coccoid) symbiotic state. SLL-2 binds with high affinity to the Forssman antigen (N-acetylgalactosamine(GalNAc)α1-3GalNAcβ1-3Galα1-4Galβ1-4Glc-ceramide), and the presence of Forssman antigen-like sugar on the surface of Symbiodinium CS-156 cells was previously confirmed. Here we report the crystal structures of SLL-2 and i...
Source: Glycobiology - July 29, 2015 Category: Biology Authors: Kita, A., Jimbo, M., Sakai, R., Morimoto, Y., Miki, K. Tags: Structural Biology Source Type: research
Globally profiling sialylation status of macrophages upon statin treatment
In this study, we performed a global profiling of the sialylation status of macrophages and their release of SAs in the cell culture medium by using flow cytometry, confocal microscopy and liquid chromatography tandem mass spectrometry (LC-MS/MS). Both flow cytometry and confocal microscopy results showed that cell surface α-2,3-linked SAs were predominant in the normal culture condition and changed slightly upon treatment with atorvastatin for 24 h, whereas α-2,6-linked SAs were negligible in the normal culture condition but significantly increased after treatment. Meanwhile, the amount of total cellular SAs i...
Source: Glycobiology - July 29, 2015 Category: Biology Authors: Wang, D., Nie, H., Ozhegov, E., Wang, L., Zhou, A., Li, Y., Sun, X.-L. Tags: Immunology Source Type: research
Human sialic acid acetyl esterase: Towards a better understanding of a puzzling enzyme
Sialic acid acetyl esterase (SIAE) removes acetyl moieties from the hydroxyl groups in position 9 and 4 of sialic acid. Recently, a dispute has been opened on its association to autoimmunity. In order to get new insights on human SIAE biology and to clarify its seemingly contradictory molecular properties, we combined in silico characterization, phylogenetic analysis and homology modeling with cellular studies in COS7 cells. Genomic and phylogenetic analysis revealed that in most tissues only the "long" isoform, originally referred to lysosomal sialic acid esterase, is detected. Using the homology modeling approach, we pre...
Source: Glycobiology - July 29, 2015 Category: Biology Authors: Orizio, F., Damiati, E., Giacopuzzi, E., Benaglia, G., Pianta, S., Schauer, R., Schwartz-Albiez, R., Borsani, G., Bresciani, R., Monti, E. Tags: Glycan Metabolism Source Type: research
Investigation of the protective properties of glycosylphosphatidylinositol-based vaccine candidates in a Toxoplasma gondii mouse challenge model
Vaccination against the ubiquitous parasite Toxoplasma gondii would provide the most efficient prevention against toxoplasmosis-related congenital, brain and eye diseases in humans. We investigated the immune response elicited by pathogen-specific glycosylphosphatidylinositol (GPI) glycoconjugates using carbohydrate microarrays in a BALB/c mouse model. We further examined the protective properties of the glycoconjugates in a lethal challenge model using the virulent T. gondii RH strain. Upon immunization, mice raised antibodies that bind to the respective GPIs on carbohydrate microarrays, but were mainly directed against a...
Source: Glycobiology - July 29, 2015 Category: Biology Authors: Gotze, S., Reinhardt, A., Geissner, A., Azzouz, N., Tsai, Y.-H., Kurucz, R., Varon Silva, D., Seeberger, P. H. Tags: Chemical Biology Source Type: research
Glycosaminoglycans affect heparanase location in CHO cell lines
Glycosaminoglycans (GAG) play a ubiquitous role in tissues and cells. In eukaryotic cells, heparan sulfate (HS) is initially degraded by an endo-β-glucuronidase called heparanase-1 (HPSE). HS oligosaccharides generated by the action of HPSE intensify the activity of signaling molecules, activating inflammatory response, tumor metastasis, and angiogenesis. The aim of the present study was to understand if sulfated GAG could modulate HPSE, since the mechanisms that regulate HPSE have not been completely defined. CHO-K1 cells were treated with 4-methylumbelliferone (4-MU) and sodium chlorate, to promote total inhibition ...
Source: Glycobiology - July 29, 2015 Category: Biology Authors: Piva, M. B., Suarez, E. R., Melo, C. M., Cavalheiro, R. P., Nader, H. B., Pinhal, M. A. Tags: Cell Biology Source Type: research
Glycosyltransferases involved in the synthesis of MUC-associated metastasis-promoting selectin ligands
In this study, quantitative real-time polymerase chain reaction analysis coupled with in situ proximity ligation assay and small interference RNA treatment shows involvement of β3galactosyltransferase-V in the synthesis of MUC16-associated sLea in H292 cells. Also, α3fucosyltransferase-V, which is absent in BEAS-2B human immortalized bronchial epithelial cells and A549 lung carcinoma cells, participates in the synthesis of MUC1-associated sLex in CFT1 human immortalized bronchial epithelial cells and H292 lung carcinoma cells. Neither selectin ligand is found on MUC1 in BEAS-2B and A549 cells. Knockdown of eithe...
Source: Glycobiology - July 29, 2015 Category: Biology Authors: Chachadi, V. B., Bhat, G., Cheng, P.-W. Tags: Cancer Biology Source Type: research
