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Alexander V Skurat, Dyann Segvich, Anna A DePaoli-Roach, Peter J Roach
Glycogen, a branched polymer of glucose, functions as an energy reserve in many living organisms. Abnormalities in glycogen metabolism, usually excessive accumulation, can be caused genetically, most often through mutation of the enzymes directly involved in synthesis and degradation of the polymer leading to a variety of glycogen storage diseases (GSDs). Microscopic visualization of glycogen deposits in cells and tissues is important for the study of normal glycogen metabolism as well as diagnosis of GSDs...
January 10, 2017: Glycobiology
Artem S Silchenko, Nadezhda E Ustyuzhanina, Mikhail I Kusaykin, Vadim B Krylov, Alexander S Shashkov, Andrey S Dmitrenok, Roza V Usoltseva, Anastasiya O Zueva, Nikolay E Nifantiev, Tatyana N Zvyagintseva
A gene that encodes fucoidanase ffa2 in the marine bacterium Formosa algae strain KMM 3553(T) was cloned, and the protein (FFA2) was produced in Escherichia coli Recombinant fucoidanase FFA2 was purified, and the biochemical properties of this enzyme were studied. The amino acid sequence of FFA2 showed 57% identity with known fucoidanase FcnA from Mariniflexile fucanivorans. The mass of the gene product FFA2 is 101.2 kDa (918 amino acid residues). Sequence analysis has revealed that fucoidanase FFA2 belongs to the GH107 (CAZy) family...
December 28, 2016: Glycobiology
Miki Tanaka-Okamoto, Mikio Mukai, Hidenori Takahashi, Yoshiyuki Fujiwara, Masayuki Ohue, Yasuhide Miyamoto
Glycomic analysis focused on sulfated O-glycans was performed to identify novel serum carbohydrate tumor markers. Sulfated glycans were enriched by α-neuraminidase digestion of pyridylaminated glycans prepared from sera, followed by anion exchange chromatography. Sulfated O-glycan profiles were constructed by two types of high performance liquid chromatography separation. Comparison of the profiles from 20 healthy controls with those of 11 gastric and 9 pancreatic cancer patients identified 14 marker candidates...
December 26, 2016: Glycobiology
Zhengliang L Wu, Xinyi Huang, Cheryl Ethen, Timothy Tatge, Marta Pasek, Joseph Zaia
Heparan sulfate (HS) is a linear polysaccharide found in the extracellular matrix (ECM) and on the cell membrane. It plays numerous roles in cellular events, including cell growth, migration and differentiation through binding to various growth factors, cytokines and other ECM proteins. Heparanase (HPSE) is an endoglycosidase that cleaves HS in the ECM and cell membrane. By degrading HS, HPSE not only alters the integrity of the ECM but also releases growth factors and angiogenic factors bound to HS chains, therefore, changes various cellular activities, including cell mobility that is critical for cancer metastasis...
December 26, 2016: Glycobiology
Mohammadreza Movahedin, Teresa M Brooks, Nitin T Supekar, Naveen Gokanapudi, Geert-Jan Boons, Cory L Brooks
In cancer cells, the glycoprotein Mucin 1 (MUC1) undergoes abnormal, truncated glycosylation. The truncated glycosylation exposes cryptic peptide epitopes that can be recognized by antibodies. Since these immunogenic regions are cancer specific, they represent ideal targets for therapeutic antibodies. We investigated the role of tumor-specific glycosylation on antigen recognition by the therapeutic antibody AR20.5. We explored the affinity of AR20.5 to a synthetic cancer-specific MUC1 glycopeptide and peptide...
December 26, 2016: Glycobiology
Daniela Nunes-Costa, Ana Maranha, Mafalda Costa, Susana Alarico, Nuno Empadinhas
Despite the progressive decline in tuberculosis mortality, strains resistant to our dated antibiotics remain a global threat, as are the emerging nontuberculous mycobacteria, ubiquitous in natural and human environments. This pressing situation boosted by debilitated immune systems, chronic illness and the aged population calls for efficient strategies to fight these successful organisms, and identifying pathways critical for their survival is a crucial step towards this goal. In this context, the glycoside glucosylglycerate (GG) has been implicated in the adaptation of mycobacteria to nitrogen starvation and to thermal stress, and the key gene for GG synthesis has been considered essential for Mycobacterium tuberculosis growth...
December 26, 2016: Glycobiology
Ksenia S Egorova, Philip V Toukach
Glycosyltransferases (GTs) are carbohydrate-active enzymes (CAZy) involved in the synthesis of natural glycan structures. The application of CAZy is highly demanded in biotechnology and pharmaceutics. However, it is being hindered by the lack of high-quality and comprehensive repositories of the research data accumulated so far. In this paper, we describe a new curated Carbohydrate Structure Glycosyltransferase Database (CSDB_GT). Currently, CSDB_GT provides ca. 780 activities exhibited by GTs, as well as several other CAZy, found in Arabidopsis thaliana and described in ca...
December 23, 2016: Glycobiology
Valentin Friedrich, Bettina Janesch, Markus Windwarder, Daniel Maresch, Matthias L Braun, Zoë A Megson, Evgeny Vinogradov, Marie-France Goneau, Ashu Sharma, Friedrich Altmann, Paul Messner, Ian C Schoenhofen, Christina Schäffer
Tannerella forsythia is an anaerobic, Gram-negative periodontal pathogen. A unique O-linked oligosaccharide decorates the bacterium's cell surface proteins and was shown to modulate the host immune response. In our study, we investigated the biosynthesis of the nonulosonic acid (NulO) present at the terminal position of this glycan. A bioinformatic analysis of T. forsythia genomes revealed a gene locus for the synthesis of pseudaminic acid (Pse) in the type strain ATCC 43037 while strains FDC 92A2 and UB4 possess a locus for the synthesis of legionaminic acid (Leg) instead...
December 16, 2016: Glycobiology
Jose O Previato, Evgeny Vinogradov, Emmanuel Maes, Leonardo M Fonseca, Yann Guerardel, Priscila A V Oliveira, Lucia Mendonça-Previato
Galactoxylomannans (GalXMs) are a mixture of neutral and acidic capsular polysaccharides produced by the human opportunistic fungus Cryptococcus neoformans that exhibit potent suppressive effects on the host immune system. Previous studies describing the chemical structure of C. neoformans GalXMs have reported species without O-acetyl substituents. Herein we describe that C. neoformans grown in capsule-inducing medium produces highly O-acetylated GalXMs. The location of the O-acetyl groups was determined by nuclear magnetic resonance (NMR) spectroscopy...
December 16, 2016: Glycobiology
Wu Di, Akiko Fujita, Kayo Hamaguchi, Philippe Delannoy, Chihiro Sato, Ken Kitajima
The occurrence and biological importance of sialic acid (Sia) and its metabolic enzymes in insect have been studied using Drosophila melanogaster The most prominent feature of D. melanogaster CMP-Sia synthetase (DmCSS) is its Golgi-localization, contrasted with nuclear localization of vertebrate CSSs. However, it remains unclear if the Golgi-localization is common to other insect CSSs and why it happens. To answer these questions, Aedes aegypti (mosquito) CSS (AaCSS) and Tribolium castaneum (beetle) CSS (TcCSS) were cloned and characterized for their activity and subcellular localization...
December 16, 2016: Glycobiology
Karin Säljö, Angela Barone, Dzeneta Vizlin-Hodzic, Bengt R Johansson, Michael E Breimer, Keiko Funa, Susann Teneberg
High expectations are held for human-induced pluripotent stem cells (hiPSC) since they are established from autologous tissues thus overcoming the risk of allogeneic immune rejection when used in regenerative medicine. However, little is known regarding the cell-surface carbohydrate antigen profile of hiPSC compared with human embryonic stem cells (hESC). Here, glycosphingolipids were isolated from an adipocyte-derived hiPSC line, and hiPSC and hESC glycosphingolipids were compared by concurrent characterization by binding assays with carbohydrate-recognizing ligands and mass spectrometry...
December 8, 2016: Glycobiology
Ken Hanzawa, Noriko Suzuki, Shunji Natsuka
Zebrafish is a model organism suitable for studying vertebrate development. We analyzed the N-glycan structures of zebrafish embryos and their alterations during zebrafish embryogenesis to obtain basic data for studying the roles of N-glycosylation. Multiple modes of high-performance liquid chromatography and multistage mass spectrometry were used for structural analysis of N-glycans. The N-glycans from deyolked embryos at 36 hours postfertilization, a mid-pharyngula stage, contained relatively higher amounts of complex- and hybrid-type glycans with LacNAc (Galβ1-4GlcNAc) and/or sialyl LacNAc without additional β1,4-Gal, which are commonly found in mammalian tissues, as well as abundant oligomannose-type glycans...
December 8, 2016: Glycobiology
Yan Liu, Ryan McBride, Mark Stoll, Angelina S Palma, Lisete Silva, Sanjay Agravat, Kiyoko F Aoki-Kinoshita, Matthew P Campbell, Catherine E Costello, Anne Dell, Stuart M Haslam, Niclas G Karlsson, Kay-Hooi Khoo, Daniel Kolarich, Milos V Novotny, Nicolle H Packer, Rene Ranzinger, Erdmann Rapp, Pauline M Rudd, Weston B Struwe, Michael Tiemeyer, Lance Wells, William S York, Joseph Zaia, Carsten Kettner, James C Paulson, Ten Feizi, David F Smith
MIRAGE (Minimum Information Required for A Glycomics Experiment) is an initiative that was created by experts in the fields of glycobiology, glycoanalytics and glycoinformatics to produce guidelines for reporting results from the diverse types of experiments and analyses used in structural and functional studies of glycans in the scientific literature. As a sequel to the guidelines for sample preparation (Struwe et al. 2016, Glycobiology, 26:907-910) and mass spectrometry  data (Kolarich et al. 2013, Mol. Cell Proteomics, 12:991-995), here we present the first version of guidelines intended to improve the standards for reporting data from glycan microarray analyses...
November 22, 2016: Glycobiology
Silvia Bleuler-Martinez, Katrin Stutz, Ramon Sieber, Mayeul Collot, Jean-Maurice Mallet, Michael Hengartner, Mario Schubert, Annabelle Varrot, Markus Künzler
Lectins are used as defense effector proteins against predators, parasites and pathogens by animal, plant and fungal innate defense systems. These proteins bind to specific glycoepitopes on the cell surfaces and thereby interfere with the proper cellular functions of the various antagonists. The exact cellular toxicity mechanism is in many cases unclear. Lectin CCL2 of the mushroom Coprinopsis cinerea was previously shown to be toxic for Caenorhabditis elegans and Drosophila melanogaster This toxicity is dependent on a single, high-affinity binding site for the trisaccharide GlcNAc(Fucα1,3)β1,4GlcNAc, which is a hallmark of nematode and insect N-glycan cores...
November 22, 2016: Glycobiology
Daisuke Sugahara, Yuka Kobayashi, Yoshihiro Akimoto, Hayato Kawakami
In this study, we examined the distribution of fucosylated glycans in mouse intestines using a lectin, BC2LCN (N-terminal domain of the lectin BC2L-C from Burkholderia cenocepacia), as a probe. BC2LCN is specific for glycans with a terminal Fucα1,2Galβ1,3-motif and it is a useful marker for discriminating the undifferentiated status of human induced/embryonic stem cells. Apparent BC2LCN reactivity was detected in the secretory granules of goblet cells in the ileum but not those in the colon. We also found distinctive reactivity in the crypt bottom, which is known as the stem cell zone, of the colon and the ileum...
November 22, 2016: Glycobiology
Kai Cheng, Yusen Zhou, Sriram Neelamegham
Glycan or carbohydrate structures can be pictorially represented using symbolic nomenclatures. The symbol nomenclature for glycans (SNFG) contains 67 different monosaccharides represented using various colors and geometric shapes. A simple tool to convert International Union of Pure and Applied Chemistry (IUPAC) format text to SNFG will be useful for sketching glycans and glycopeptides. Such code can also enable the development of more sophisticated applications, where the visual representation of carbohydrate structures is necessary...
November 22, 2016: Glycobiology
Ryan J Blackler, Susannah M L Gagnon, Robert Polakowski, Natisha L Rose, Ruixiang B Zheng, James A Letts, Asha R Johal, Brock Schuman, Svetlana N Borisova, Monica M Palcic, Stephen V Evans
The homologous glycosyltransferases α-1,3-N-acetylgalactosaminyltransferase (GTA) and α-1,3-galactosyltransferase (GTB) carry out the final synthetic step of the closely related human ABO(H) blood group A and B antigens. The catalytic mechanism of these model retaining enzymes remains under debate, where Glu303 has been suggested to act as a putative nucleophile in a double displacement mechanism, a local dipole stabilizing the intermediate in an orthogonal associative mechanism or a general base to stabilize the reactive oxocarbenium ion-like intermediate in an S N i-like mechanism...
November 22, 2016: Glycobiology
María Florencia Festari, Felipe Trajtenberg, Nora Berois, Sergio Pantano, Leslie Revoredo, Yun Kong, Patricia Solari-Saquieres, Yoshiki Narimatsu, Teresa Freire, Sylvie Bay, Carlos Robello, Jean Bénard, Thomas A Gerken, Henrik Clausen, Eduardo Osinaga
Polypeptide GalNAc-transferases (GalNAc-Ts) constitute a family of 20 human glycosyltransferases (comprising 9 subfamilies), which initiate mucin-type O-glycosylation. The O-glycoproteome is thought to be differentially regulated via the different substrate specificities and expression patterns of each GalNAc-T isoforms. Here, we present a comprehensive in vitro analysis of the peptide substrate specificity of GalNAc-T13, showing that it essentially overlaps with the ubiquitous expressed GalNAc-T1 isoform found in the same subfamily as T13...
November 22, 2016: Glycobiology
M Osman Sheikh, Stephanie M Halmo, Sneha Patel, Dustin Middleton, Hideyuki Takeuchi, Christopher M Schafer, Christopher M West, Robert S Haltiwanger, Fikri Y Avci, Kelley W Moremen, Lance Wells
Determining the correct enzymatic activity of putative glycosyltransferases (GTs) can be challenging as these enzymes can utilize multiple donor and acceptor substrates. Upon initial determination of the donor-sugar nucleotide(s), a GT utilizes various acceptor molecules that can then be tested. Here, we describe a quick method to screen sugar-nucleotide donor specificities of GTs utilizing a sensitive, nonradioactive, commercially available bioluminescent uridine diphosphate detection kit. This in vitro method allowed us to validate the sugar-nucleotide donor-substrate specificities of recombinantly expressed human, bovine, bacterial and protozoan GTs...
November 22, 2016: Glycobiology
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October 31, 2016: Glycobiology
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