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Hideyuki Takeuchi, Derek Wong, Michael Schneider, Hudson H Freeze, Megumi Takeuchi, Steven J Berardinelli, Atsuko Ito, Hane Lee, Stanley F Nelson, Robert S Haltiwanger
Protein O-fucosyltransferase-1 (POFUT1) adds O-fucose monosaccharides to epidermal growth factor-like (EGF) repeats found on approximately 100 mammalian proteins, including Notch receptors. Haploinsufficiency of POFUT1 has been linked to adult-onset Dowling Degos Disease (DDD) with hyperpigmentation defects. Homozygous deletion of mouse Pofut1 results in embryonic lethality with severe Notch-like phenotypes including defects in somitogenesis, cardiogenesis, vasculogenesis, and neurogenesis, but the extent to which POFUT1 is required for normal human development is not yet understood...
February 14, 2018: Glycobiology
Miki Tanaka-Okamoto, Ken Hanzawa, Mikio Mukai, Hidenori Takahashi, Masayuki Ohue, Yasuhide Miyamoto
In our previous study, 14 sulfated carbohydrate tumor marker candidates were identified by focused glycomic analyses. Here, glycomic analyses focused on internally sialylated glycans to identify novel marker candidates. Internally sialylated glycans were enriched by digestion of pyridylaminated glycans prepared from sera withα-neuraminidase from Salmonella typhimurium, which did not cleave sialic acids linked to internal residues, followed by anion-exchange chromatography. Next, internally sialylated O-glycan profiles were constructed using two types of high performance liquid chromatography, which were compared between 20 healthy controls and 11 patients with gastric cancer and 9 patients with pancreatic cancer...
January 30, 2018: Glycobiology
Matthew R Kudelka, Alison V Nairn, Mohammed Y Sardar, Xiaodong Sun, Elliot L Chaikof, Tongzhong Ju, Kelley W Moremen, Richard D Cummings
Mucin-type O-glycans decorate >80% of secretory and cell surface proteins and contribute to health and disease. However, dynamic alterations in the O-glycome are poorly understood because current O-glycomic methodologies are not sufficiently sensitive nor quantitative. Here we describe a novel isotope labeling approach termed Isotope-Cellular O-glycome Reporter Amplification (ICORA) to amplify and analyze the O-glycome from cells. In this approach, cells are incubated with Ac3GalNAc-Bn (Ac3GalNAc-[1H7]Bn) or a heavy labeled Ac3GalNAc-BnD7 (Ac3GalNAc-[2D7]Bn) O-glycan precursor (7 Da mass difference), which enters cells and upon de-esterification is modified by Golgi enzymes to generate Bn-O-glycans secreted into the culture media...
January 30, 2018: Glycobiology
Yanxin Wang, Xin Niu, Xiaoli Guo, Han Yu, Zhonghua Liu, Zhenqing Zhang, Sheng Yuan
Two chitin deacetylases, Cda1 and Cda2, from C. cinerea were expressed and characterized. Cda1 preferably deacetylates the non-reducing end residue of (GlcNAc)2, the internal or non-reducing end residue of (GlcNAc)3, and the non-reducing residue of (GlcNAc)6 after deacetylating the internal residues. In contrast, Cda2 preferably deacetylates the reducing end residue of (GlcNAc)2, the internal or reducing end residue of (GlcNAc)3, and the reducing residue of (GlcNAc)6 after deacetylating the internal residues...
January 23, 2018: Glycobiology
Catharina Steentoft, Denis Migliorini, Tiffany R King, Ulla Mandel, Carl H June, Avery D Posey
Cancer immunotherapy is rapidly advancing in the treatment of a variety of hematopoietic cancers, including pediatric acute lymphoblastic leukemia and diffuse large B cell lymphoma, with chimeric antigen receptor (CAR)-T cells. CARs are genetically encoded artificial T cell receptors that combine the antigen specificity of an antibody with the machinery of T cell activation. However, implementation of CAR technology in the treatment of solid tumors has been progressing much slower. Solid tumors are characterized by a number of challenges that need to be overcome, including cellular heterogeneity, immunosuppressive tumor microenvironment (TME), and, in particular, few known cancer-specific targets...
January 23, 2018: Glycobiology
Nadeem Ahmad Ansari, Dharmendra Kumar Chaudhary, Debabrata Dash
Background: Dicarbonyl compounds react more rapidly, than glucose, with arginine and lysine in proteins to form advanced glycation end products (AGEs) and further produce free radicals which cause DNA damage. AGEs are reliable diagnostic biomarkers for most of the age-related diseases. Methods: In the present study histone was modified with glyoxal and it was characterized by various spectral techniques. Binding characteristics of the modified histone towards serum antibodies from type 1 diabetes patients was evaluated by solid phase enzyme immunoassay and the results were compared with normal human subjects...
January 19, 2018: Glycobiology
Bruce Caterson, James Melrose
From an evolutionary perspective keratan sulphate (KS) is the newest glycosaminoglycan (GAG) but the least understood. KS is a sophisticated molecule with a diverse structure, and unique functional roles continue to be uncovered for this GAG. The cornea is the richest tissue source of KS in the human body but the central and peripheral nervous systems also contain significant levels of KS and a diverse range of KS-proteoglycans with essential functional roles. KS also displays important cell regulatory properties in epithelial and mesenchymal tissues and in bone and in tumour development of diagnostic and prognostic utility...
January 11, 2018: Glycobiology
Adrian J Jervis, Alison G Wood, Joel A Cain, Jonathan A Butler, Helen Frost, Elizabeth Lord, Rebecca Langdon, Stuart J Cordwell, Brendan W Wren, Dennis Linton
N-linked protein glycosylation systems operate in species from all three domains of life. The model bacterial N-linked glycosylation system from Campylobacter jejuni is encoded by pgl genes present at a single chromosomal locus. This gene cluster includes the pglB oligosaccharyltransferase responsible for transfer of glycan from lipid carrier to protein. Although all genomes from species of the Campylobacter genus contain a pgl locus, among the related Helicobacter genus only three evolutionarily related species (H...
January 11, 2018: Glycobiology
Ewa Jankowska, Lisa M Parsons, Xuezheng Song, Dave F Smith, Richard D Cummings, John F Cipollo
Here we present a Caenorhabditis elegans N-glycan shotgun array. This nematode serves as a model organism for many areas of biology including but not limited to tissue development, host-pathogen interactions, innate immunity, and genetics. C. elegans N-glycans contain structural motifs that are also found in other nematodes as well as trematodes and lepidopteran species. Glycan binding toxins that interact with C. elegans glycoconjugates also do so with some agriculturally relevant species, such as Haemonchus contortus, Ascaris suum, Oesophagostomum dentatum and Trichoplusia ni...
January 9, 2018: Glycobiology
Pratima Nangia-Makker, Victor Hogan, Avraham Raz
Over the last few decades galectin-3, a carbohydrate binding protein, with affinity for N-acetyllactosamine residues, has been unique due to the regulatory roles it performs in processes associated with tumor progression and metastasis such as cell proliferation, homotypic/ heterotypic aggregation, dynamic cellular transformation, migration and invasion, survival and apoptosis. Structure-function association of galectin-3 reveals that it consists of a short amino terminal motif, which regulates its nuclear-cytoplasmic shuttling; a collagen α-like domain, susceptible to cleavage by matrix metalloproteases and prostate specific antigen (PSA); accountable for its oligomerization and lattice formation, and a carbohydrate recognition/binding domain containing the anti-death motif of the Bcl2 protein family...
January 5, 2018: Glycobiology
Yoshiki Narimatsu, Hiren J Joshi, Yang Zhang, Catarina Gomes, Yen-Hsi Chen, Flaminia Lorenzetti, Sanae Furukawa, Katrine Schjoldager, Lars Hansen, Henrik Clausen, Eric P Bennett, Hans H Wandall
Over 200 glycosyltransferases are involved in the orchestration of the biosynthesis of the human glycome , which is comprised of all glycan structures found on different glycoconjugates in cells. The glycome is vast, and despite advancements in analytic strategies it continues to be difficult to decipher biological roles of glycans with respect to specific glycan structures, type of glycoconjugate, particular glycoproteins, and distinct glycosites on proteins. In contrast to this, the number of glycosyltransferase genes involved in the biosynthesis of the human glycome is manageable, and the biosynthetic roles of most of these enzymes are defined or can be predicted with reasonable confidence...
January 5, 2018: Glycobiology
Eva Bönisch, Yoo Jin Oh, Julia Anzengruber, Fiona F Hager, Arturo Lopez-Guzman, Sonja Zayni, Peter Hinterdorfer, Paul Kosma, Paul Messner, Katarzyna A Duda, Christina Schäffer
The Gram-positive lactic acid bacterium Lactobacillus buchneri CD034 is covered by a two-dimensional crystalline, glycoproteinaceous cell surface (S-) layer lattice. While lactobacilli are extensively exploited as cell surface display systems for applied purposes, questions about how they stick their cell wall together are remaining open. This also includes the identification of the S-layer cell wall ligand. In this study, lipoteichoic acid was isolated from the L. buchneri CD034 cell wall as a significant fraction of the bacterium's cell wall glycopolymers, structurally characterized and analyzed for its potential to mediate binding of the S-layer to the cell wall...
January 4, 2018: Glycobiology
Xiayun Huang, Tannin A Schmidt, Claire Shortt, Shivani Arora, Akira Asari, Thorsten Kirsch, Mary K Cowman
A method for specific quantification of hyaluronan (HA) concentration using AlphaScreen® (Amplified Luminescent Proximity Homogeneous Assay) technology is described. Two types of hydrogel-coated and chromophore-loaded latex nanobeads are employed. The proximity of the beads in solution is detected by excitation of the donor bead leading to the production of singlet oxygen, and chemiluminescence from the acceptor bead upon exposure to singlet oxygen. In the HA assay, the donor bead is modified with streptavidin, and binds biotin-labeled HA...
December 29, 2017: Glycobiology
Jiyong Su, Jin Gao, Yunlong Si, Linlin Cui, Chenyang Song, Yue Wang, Runjie Wu, Guihua Tai, Yifa Zhou
Galectin-10 (Gal-10) which forms Charcot-Leyden crystals in vivo, is crucial to regulating lymph cell function. Here, we solved the crystal structures of Gal-10 and eight variants at resolutions of 1.55-2.00 Å. Structural analysis and size exclusion chromatography demonstrated that Gal-10 dimerizes with a novel global shape that is different from that of other prototype galectins (e.g., Gal-1, -2 and -7). In the Gal-10 dimer, Glu33 from one subunit modifies the carbohydrate binding site of another, essentially inhibiting disaccharide binding...
December 23, 2017: Glycobiology
Olivia Joan Adams, Michal A Stanczak, Stephan von Gunten, Heinz Läubli
Changes in sialic acids in cancer have been observed for many years. In particular, the increase of sialoglycan density or hypersialylation in tumors has been described. Recent studies have identified mechanisms for immune evasion based on sialoglycan interactions with immunoregulatory Siglec receptors that are exploited by tumor cells and microorganisms alike. Siglecs are mostly inhibitory receptors similar to known immune checkpoints including PD-1 or CTLA-4 that are successfully targeted with blocking antibodies for cancer immunotherapy...
December 22, 2017: Glycobiology
Olaf Perdijk, R J Joost van Neerven, Ben Meijer, Huub F J Savelkoul, Sylvia Brugman
The human milk oligosaccharide 3'sialyllactose (3'SL) has previously been shown to activate murine dendritic cells (DC) in a TLR4-mediated manner ex vivo. In this study we aimed to investigate whether 3'SL has similar immunomodulatory properties on human DC. 3'SL was shown to induce NF-κB activation via human TLR4. However, LPS was detected in the commercially obtained 3'SL from different suppliers. After the removal of LPS from 3'SL, we studied its ability to modify DC differentiation in vitro. In contrast to LPS and 3'SL, LPS-free 3'SL did not induce functional and phenotypical changes on immature DC (iDC)...
December 21, 2017: Glycobiology
Lubor Borsig
Selectins are vascular adhesion molecules that mediate physiological responses such as inflammation, immunity and hemostasis. During cancer progression selectins promote various steps enabling the interactions between tumor cells and the blood constituents, including platelets, endothelial cells, and leukocytes. Selectins are carbohydrate-binding molecules that bind to sialylated, fucosylated glycan structures. The increased selectin ligand expression on tumor cells correlates with enhanced metastasis and poor prognosis for cancer patients...
December 20, 2017: Glycobiology
Hiren J Joshi, Anja Jørgensen, Katrine T Schjoldager, Adnan Halim, Leo A Dworkin, Catharina Steentoft, Hans H Wandall, Henrik Clausen, Sergey Y Vakhrushev
The GlycoDomainViewer is a bioinformatic tool to aid in the mining of glycoproteomic data sets from different sources and facilitate incorporation of glycosylation into studies of protein structure and function. We present a version 2.0 of GlycoDomainViewer incorporating a number of advanced features, which enhances visibility and accessibility of the wealth of glycoproteomic data being generated. The GlycoDomainViewer enables visual exploration of glycoproteomic data, incorporating information from recent N- and O-glycoproteome studies on human and animal cell lines and some organs and body fluids...
December 18, 2017: Glycobiology
Shonoi A Ming, Ebony Cottman-Thomas, Natalee C Black, Yi Chen, Vamsee Veeramachineni, Dwight C Peterson, Xi Chen, Lauren M Tedaldi, Gerd K Wagner, Chao Cai, Robert J Linhardt, Willie F Vann
Neisseria meningitidis Group X is an emerging cause of bacterial meningitis in the Sub-Saharan Africa. The capsular polysaccharide of Group X is a homopolymer of N-acetylglucosamine α(1-4) phosphate and is a vaccine target for prevention of disease associated with this meningococcal serogroup. We have demonstrated previously that the formation of the polymer is catalyzed by a phosphotransferase which transfers N- acetylglucosamine-1-phosphate from UDP-N-acetylglucosamine to the 4- hydroxyl of the N-acetylglucosamine on the non-reducing end of the growing chain...
December 8, 2017: Glycobiology
Takahiro Yoshizawa, Shuji Mizumoto, Yuki Takahashi, Shin Shimada, Kazuyuki Sugahara, Jun Nakayama, Shin'ichi Takeda, Yoshihiro Nomura, Yuko Nitahara-Kasahara, Takashi Okada, Kiyoshi Matsumoto, Shuhei Yamada, Tomoki Kosho
Collagen is one of the most important components of the extracellular matrix that is involved in the strength of tissues, cell adhesion, and cell proliferation. Mutations in several collagen and post-translational modification enzyme genes cause Ehlers-Danlos syndrome (EDS) characterized by joint and skin hyperextensibility as well as fragility of various organs. Carbohydrate sulfotransferase 14/dermatan 4-O-sulfotransferase-1 (CHST14/D4ST1) is a critical enzyme for biosynthesis of dermatan sulfate, a side chain of various proteoglycans including biglycan that regulates collagen fibrils through their interaction...
December 1, 2017: Glycobiology
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