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Yoshiyuki Goto, Satoshi Uematsu, Hiroshi Kiyono
Intestinal epithelial cells apically express glycans, especially α1,2-fucosyl linkages, which work as a biological interface for the host-microbe interaction. Emerging studies have shown that epithelial α1,2-fucosylation is regulated by microbes and by group 3 innate lymphoid cells (ILC3s). Dysregulation of the gene (FUT2) encoding fucosyltransferase 2, an enzyme governing epithelial α1,2-fucosylation, is associated with various human disorders, including infection and chronic inflammatory diseases. This suggests a critical role for an interaction between microbes, epithelial cells and ILC3s mediated via glycan residues...
October 19, 2016: Nature Immunology
Faisal Aziz, Wei Gao, Qiu Yan
H. pylori cytotoxin associated antigen A (CagA) plays a significant role in the progression of gastric cancer but their effect on fucosylation to develop gastric cancer is unknown. Fucosyltransferase IV (FUT4) is the key enzyme for synthesis of LewisY (LeY) carried by glycoproteins and glycolipids on the cell membrane. Herein, we compare the expression of CagA, p-EGFR, FUT4 and LeY in gastritis (n = 128, 176), gastric ulcer (n = 174, 213), and gastric cancer (n = 323, 261) tissue and serum samples, respectively by IHC and ELISA...
October 18, 2016: Pathology Oncology Research: POR
Daniel Garrido, Santiago Ruiz-Moyano, Nina Kirmiz, Jasmine C Davis, Sarah M Totten, Danielle G Lemay, Juan A Ugalde, J Bruce German, Carlito B Lebrilla, David A Mills
The infant intestinal microbiota is often colonized by two subspecies of Bifidobacterium longum: subsp. infantis (B. infantis) and subsp. longum (B. longum). Competitive growth of B. infantis in the neonate intestine has been linked to the utilization of human milk oligosaccharides (HMO). However, little is known how B. longum consumes HMO. In this study, infant-borne B. longum strains exhibited varying HMO growth phenotypes. While all strains efficiently utilized lacto-N-tetraose, certain strains additionally metabolized fucosylated HMO...
October 19, 2016: Scientific Reports
Shuxin Shang, Lu Sun, Wei Li, Xue Qin, Shu Zhang, Yinkun Liu
Fucosylated glycans play a critical role in cell biology processes during the occurrence and development of tumor, such as cell signal transduction, cellular differentiation, ligand-receptor interactions, and metastasis formation. Fucosylated haptoglobin (Hp) was reported to be increased in hepatocellular carcinoma (HCC). In this study, we aimed to develop a convenient and sensitive AAL@Magbeads-based Hp ELISA assay to rapidly detect fucosylated Hp in 270 serum samples from HCC patients and normal controls...
September 2016: Discovery Medicine
Carmen Jiménez-Castells, Jorick Vanbeselaere, Sonja Kohlhuber, Bärbel Ruttkowski, Anja Joachim, Katharina Paschinger
BACKGROUND: The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. METHODS: Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae...
October 14, 2016: Biochimica et Biophysica Acta
Karin Förster-Fromme, Sarah Schneider, Georg A Sprenger, Christoph Albermann
OBJECTIVES: To investigate the translocation of nucleotide-activated sugars from the cytosol across a membrane into the endoplasmatic reticulum or the Golgi apparatus which is an important step in the synthesis of glycoproteins and glycolipids in eukaryotes. RESULTS: The heterologous expression of the recombinant and codon-adapted human GDP-L-fucose antiporter gene SLC35C1 (encoding an N-terminal OmpA-signal sequence) led to a functional transporter protein located in the cytoplasmic membrane of Escherichia coli...
October 13, 2016: Biotechnology Letters
Tomasz Walski, Els J M Van Damme, Nicolas Smargiasso, Olivier Christiaens, Edwin De Pauw, Guy Smagghe
In holometabolous insects the transition from larva to adult requires a complete body reorganization and relies on N-glycosylated proteins. N-glycosylation is an important posttranslational modification that influences protein activity but its impact on the metamorphosis has not been studied yet. Here we used the red flour beetle, Tribolium castaneum, to perform a first comprehensive study on the involvement of the protein N-glycosylation pathway in metamorphosis. The transcript levels for genes encoding N-glycan processing enzymes increased during later developmental stages and, in turn, transition from larva to adult coincided with an enrichment of more extensively modified paucimannose glycans, including fucosylated ones...
October 12, 2016: Scientific Reports
Sung Hyeon Lee, Seunghyup Jeong, Jua Lee, In Seok Yeo, Myung Jin Oh, Unyong Kim, Sumin Kim, Su Hee Kim, Seung-Yeol Park, Jae-Han Kim, Se Hoon Park, Jung Hoe Kim, Hyun Joo An
Gastric cancer has one of the highest cancer mortality rates worldwide, largely because of difficulties in early-stage detection. Aberrant glycosylation in serum proteins is associated with many human diseases including inflammation and various types of cancer. Serum-based global glycan profiling using mass spectrometry has been explored and has already led to several potential glycan markers for several disease states. However, localization of the aberrant glycosylation is desirable in order to improve the specificity and sensitivity for clinical use...
October 5, 2016: Molecular BioSystems
Anna Sroka-Bartnicka, Isabella Karlsson, Lorena Ndreu, Alessandro Quaranta, Matthijs Pijnappel, Gunnar Thorsén
Glycosylation is one of the most common and important post-translational modifications, influencing both the chemical and the biological properties of proteins. Studying the glycosylation of the entire protein population of a sample can be challenging because variations in the concentrations of certain proteins can enhance or obscure changes in glycosylation. Furthermore, alterations in the glycosylation pattern of individual proteins, exhibiting larger variability in disease states, have been suggested as biomarkers for different types of cancer, as well as inflammatory and neurodegenerative diseases...
October 3, 2016: Journal of Pharmaceutical and Biomedical Analysis
Anna Kałuża, Anna Jarząb, Andrzej Gamian, Ewa M Kratz, Mariusz Zimmer, Mirosława Ferens-Sieczkowska
Glycosylation pattern within reproductive tract is now suggested to be involved in providing female immune tolerance for allograft sperm and developing embryo, but the information whether impaired glycosylation may influence male fertility potential is still limited. We have analyzed seminal plasma N-glycome in pooled samples derived from fertile and infertile men by means of MALDI-TOF/TOF tandem mass spectrometry. Among infertile subjects, normozoospermic, oligozoospermic, asthenozoospermic and oligoasthenozoospermic samples were obtained...
September 18, 2016: Carbohydrate Research
Johanne Dubail, Deepika Vasudevan, Lauren W Wang, Sarah E Earp, Michael W Jenkins, Robert S Haltiwanger, Suneel S Apte
Peters Plus syndrome (PPS), a congenital disorder of glycosylation, results from recessive mutations affecting the glucosyltransferase B3GLCT, leading to congenital corneal opacity and diverse extra-ocular manifestations. Together with the fucosyltransferase POFUT2, B3GLCT adds Glucoseβ1-3Fucose disaccharide to a consensus sequence in thrombospondin type 1 repeats (TSRs) of several proteins. Which of these target proteins is functionally compromised in PPS is unknown. We report here that haploinsufficiency of murine Adamts9, encoding a secreted metalloproteinase with 15 TSRs, leads to congenital corneal opacity and Peters anomaly (persistent lens-cornea adhesion), which is a hallmark of PPS...
September 30, 2016: Scientific Reports
Wan-Ling Ho, Wen-Ming Hsu, Min-Chuan Huang, Kenji Kadomatsu, Akira Nakagawara
Glycosylation is the most complex post-translational modification of proteins. Altered glycans on the tumor- and host-cell surface and in the tumor microenvironment have been identified to mediate critical events in cancer pathogenesis and progression. Tumor-associated glycan changes comprise increased branching of N-glycans, higher density of O-glycans, generation of truncated versions of normal counterparts, and generation of unusual forms of terminal structures arising from sialylation and fucosylation. The functional role of tumor-associated glycans (Tn, sTn, T, and sLe(a/x)) is dependent on the interaction with lectins...
September 29, 2016: Journal of Hematology & Oncology
Saulius Vainauskas, Rebecca M Duke, James McFarland, Colleen McClung, Cristian Ruse, Christopher H Taron
A novel fucose-binding lectin (SL2-1) from the bacterium Streptomyces rapamycinicus was identified by analysis of metagenomic DNA sequences. SL2-1 belongs to a new group of bacterial fucose-specific lectins that have no similarity to known bacterial fucose-binding proteins, but are related to certain eukaryotic fucose-binding lectins. The 17 kDa protein was expressed recombinantly in E. coli and purified by affinity chromatography. Glycan microarray analysis with fluorescently labeled recombinant SL2-1 demonstrated its ability to bind to core α1-6 fucosylated N-glycans, but not to core α1-3 fucosylated N-glycans, or other α1-2, α1-3 and α1-4 fucosylated oligosaccharides...
September 28, 2016: Scientific Reports
Benedetta Bertolotti, Beáta Oroszová, Ieva Sutkeviciute, Ladislav Kniežo, Franck Fieschi, Kamil Parkan, Zuzana Lovyová, Martina Kašáková, Jitka Moravcová
The discovery of effective ligands for DC-SIGN receptor is one of the most challenging concepts of antiviral drug design due to the importance of this C-type lectin in infection processes. DC-SIGN recognizes mannosylated and fucosylated oligosaccharides but glycosidic linkages are accessible to both chemical and enzymatic degradations. To avoid this problem, the synthesis of stable glycoside mimetics has attracted increasing attention. In this work we establish for the first time mono- and divalent C-glycosides based on d-manno and l-fuco configurations as prospective DC-SIGN ligands...
September 9, 2016: Carbohydrate Research
Salina Louie, Benjamin Haley, Brett Marshall, Amy Heidersbach, Mandy Yim, Martina Brozynski, Danming Tang, Cynthia Lam, Bronislawa Petryniak, David Shaw, Jeongsup Shim, Aaron Miller, John B Lowe, Brad Snedecor, Shahram Misaghi
During antibody dependent cell cytotoxicity (ADCC) the target cells are killed by monocytes and natural killer cells. ADCC is enhanced when the antibody heavy chain's core N-linked glycan lacks the fucose molecule(s). Several strategies have been utilized to generate fully afucosylated antibodies. A commonly used and efficient approach has been knocking out the FUT8 gene of the Chinese hamster ovary (CHO) host cells, which results in expression of antibody molecules with fully afucosylated glycans. However, a major drawback of the FUT8-KO host is the requirement for undertaking two separate cell line development (CLD) efforts in order to obtain both primarily fucosylated and fully afucosylated antibody species for comparative studies in vitro and in vivo...
September 26, 2016: Biotechnology and Bioengineering
Bas C Jansen, Albert Bondt, Karli R Reiding, Sicco A Scherjon, Gestur Vidarsson, Manfred Wuhrer
During pregnancy, the mother provides multiple nutrients and substances to the foetus, with maternal immunoglobulin G (IgG) being actively transported to the foetus. Newborns depend on maternal IgG for immune-protection in their first months. The glycosylation of IgG has been shown to influence its dynamics, e.g. receptor binding. While minor differences in IgG glycosylation have been found between IgG derived from maternal blood and umbilical cord blood (UC) of newborn children, the differential glycosylation of maternal and UC plasma has hitherto not been studied...
September 26, 2016: Scientific Reports
Giulia Bandini, John R Haserick, Edwin Motari, Dinkorma T Ouologuem, Sebastian Lourido, David S Roos, Catherine E Costello, Phillips W Robbins, John Samuelson
Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. Although gene regulation is important for parasite differentiation and pathogenesis, little is known about protein organization in the nucleus. Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. AAL also binds to Hammondia and Neospora nuclei but not to more distantly related apicomplexans...
October 11, 2016: Proceedings of the National Academy of Sciences of the United States of America
Montserrat Mancera-Arteu, Estela Giménez, José Barbosa, Victòria Sanz-Nebot
In this study, a ZIC-HILIC-MS methodology for the analysis of N-glycan isomers was optimized to obtain greater detection sensitivity and thus identify more glycan structures in hAGP. In a second step, this method was combined with glycan reductive isotope labelling (GRIL) through [(12)C6]/[(13)C6]-aniline and exoglycosidase digestion to characterize the different glycan isomers. The GRIL method allows the peak areas resulting from two different labelled samples to be compared, since neither retention time shifts nor variations in the ionization of glycans between these samples are obtained...
October 12, 2016: Analytica Chimica Acta
Austin T Mudd, Jaime Salcedo, Lindsey S Alexander, Stacey K Johnson, Caitlyn M Getty, Maciej Chichlowski, Brian M Berg, Daniela Barile, Ryan N Dilger
BACKGROUND: Milk oligosaccharides (OSs) are bioactive components known to influence neonatal development. These compounds have specific physiological functions acting as prebiotics, immune system modulators, and enhancing intestine and brain development. OBJECTIVES: The pig is a commonly used model for studying human nutrition, and there is interest in quantifying OS composition of porcine milk across lactation compared with human milk. In this study, we hypothesized that OS and sialic acid (SA) composition of porcine milk would be influenced by stage of lactation...
2016: Frontiers in Nutrition
Pamela Norton, Mary Ann Comunale, Harmin Herrera, Mengjun Wang, Josef Houser, Michaela Wimmerova, Patrick R Romano, Anand Mehta
The Aleuria aurantia lectin (AAL) derived from orange peel fungus contains five fucose-binding sites that recognizes fucose bound in α-1,2, α-1,3, α-1,4 and α-1,6 linkages to N-acetylglucosamine (GlcNAc) and galactose. Recently, we have created several recombinant AAL (rAAL) proteins that had altered binding affinity to fucose linkages. In this report we further characterize the binding specificity of one of the mutated lectins, N224Q lectin. This lectin was characterized by lectin western blotting, surface plasmon resonance and glycan microarray and shown to have increased binding to fucosylated glycan...
September 21, 2016: Proteomics
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