ALBERT

Description:    High-mannose type N-linked glycan with 6 mannosyl residues, termed "M6Gn2", displayed clear binding to the same M6Gn2, conjugated with ceramide mimetic (cer-m) and incorporated in liposome, or coated on polystyrene plates. However, the conjugate of M6Gn2-cer-m did not interact with complex-type N-linked glycan with various structures having multiple GlcNAc termini, conjugated with cer-m. The following observations indicate that hamster embryonic fibroblast NIL-2 K cells display homotypic autoadhesion, mediated through the self-recognition capability of high-mannose type glycans expressed on these cells: (i) NIL-2 K cells display clear binding to lectins capable of binding to high-mannose type glycans ( e.g. , ConA), but not to other lectins capable of binding to other carbohydrates ( e.g. GS-II). (ii) NIL-2 K cells adhere strongly to plates coated with M6Gn2-cer-m, but not to plates coated with complex-type N-linked glycans having multiple GlcNAc termini, conjugated with cer-m; (iii) degree of NIL-2 K cell adhesion to plates coated with M6Gn2-cer-m showed a clear dose-dependence on the amount of M6Gn2-cer-m; and (iv) the degree of NIL-2 K adhesion to plates coated with M6Gn2-cer-m was inhibited in a dose-dependent manner by α1,4-L-mannonolactone, the specific inhibitor in high-mannose type glycans addition. These data indicate that adhesion of NIL-2 K is mediated by self-aggregation of high mannose type glycan. Further studies are to be addressed on auto-adhesion of other types of cells based on self interaction of high mannose type glycans. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9449-3 Authors Seon-Joo Yoon, Division of Biomembrane Research, Pacific Northwest Research Institute, and Department of Global Health, University of Washington, Seattle, WA 98122, USA Natalia Utkina, Division of Biomembrane Research, Pacific Northwest Research Institute, and Department of Global Health, University of Washington, Seattle, WA 98122, USA Martin Sadilek, Depart of Chemistry, University of Washington, Seattle, WA 98195, USA Hirokazu Yagi, Graduate School of Pharmaceutical Sciences, Nagoya City University, Tanabe-dori 3-1, Mizuho-ku, Nagoya, 467-8603 Japan Koichi Kato, Graduate School of Pharmaceutical Sciences, Nagoya City University, Tanabe-dori 3-1, Mizuho-ku, Nagoya, 467-8603 Japan Sen-itiroh Hakomori, Division of Biomembrane Research, Pacific Northwest Research Institute, and Department of Global Health, University of Washington, Seattle, WA 98122, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Human alpha-1-antitrypsin (α1AT) is a glycoprotein with protease inhibitor activity protecting tissues from degradation. Patients with inherited α1AT deficiency are treated with native α1AT (nAT) purified from human plasma. In the present study, recombinant α1AT (rAT) was produced in Chinese hamster ovary (CHO) cells and their glycosylation patterns, inhibitory activity and in vivo half-life were compared with those of nAT. A peptide mapping analysis employing a deglycosylation reaction confirmed full occupancy of all three glycosylation sites and the equivalency of rAT and nAT in terms of the protein level. N -glycan profiles revealed that rAT contained 10 glycan structures ranging from bi-antennary to tetra-antennary complex-type glycans while nAT displayed six peaks comprising majorly bi-antennary glycans and a small portion of tri-antennary glycans. In addition, most of the rAT glycans were shown to have only core α(1 - 6)-fucose without terminal fucosylation, whereas only minor portions of the nAT glycans contained core or Lewis X-type fucose. As expected, all sialylated glycans of rAT were found to have α(2 - 3)-linked sialic acids, which was in sharp contrast to those of nAT, which had mostly α(2 - 6)-linked sialic acids. However, the degree of sialylation of rAT was comparable to that of nAT, which was also supported by an isoelectric focusing gel analysis. Despite the differences in the glycosylation patterns, both α1ATs showed nearly equivalent inhibitory activity in enzyme assays and serum half-lives in a pharmacokinetic experiment. These results suggest that rAT produced in CHO cells would be a good alternative to nAT derived from human plasma. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9453-7 Authors Kyung Jin Lee, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Sang Mee Lee, Alteogen Inc., Bioventure town, Daejeon, 305-812 Korea Jin Young Gil, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Ohsuk Kwon, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Jin Young Kim, Korea Basic Science Institute, Ochang-eup, Cheongwon-gun, Chungbuk 363-883, Korea Soon Jae Park, Alteogen Inc., Bioventure town, Daejeon, 305-812 Korea Hye-Shin Chung, Alteogen Inc., Bioventure town, Daejeon, 305-812 Korea Doo-Byoung Oh, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 305-806 Korea Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    As one of several biologically active compounds in milk, glycoproteins have been indicated to be involved in the protection of newborns from bacterial infection. As much of the physical and immune development of the tammar wallaby ( Macropus eugenii ) young occurs during the early phases of lactation and not in utero , the tammar is a model species for the characterization of potential developmental support agents provided by maternal milk. In the present study, the N - and O -linked glycans from tammar wallaby milk glycoproteins from six individuals at different lactation time points were subjected to glycomics analyses using porous graphitized carbon liquid chromatography electrospray ionization mass spectrometry. Structural characterization identified a diverse range of glycan structures on wallaby milk glycoproteins including sialylated, sulphated, core fucosylated and O -fucosylated structures. 30 % of N -linked structures contained a core (α1-6) fucose. Several of these structures may play roles in development, and exhibit statistically significant temporal changes over the lactation period. The N -glycome was found to contain structures with NeuGc residues, while in contrast the O -glycome did not. O -fucosylated structures were identified in the early stages of lactation indicating a potential role in the early stages of development of the pouch young. Overall the results suggest that wallaby milk contains structures known to have developmental and immunological significance in human milk and reproduction in other animals, highlighting the importance of glycoproteins in milk. Content Type Journal Article Pages 1-14 DOI 10.1007/s10719-012-9452-8 Authors Katherine Wongtrakul-Kish, Biomolecular Frontiers Research Centre, Department of Chemistry and Biomolecular Sciences, Macquarie University, Building E8C Room 307, North Ryde, NSW 2109, Australia Daniel Kolarich, Biomolecular Frontiers Research Centre, Department of Chemistry and Biomolecular Sciences, Macquarie University, Building E8C Room 307, North Ryde, NSW 2109, Australia Dana Pascovici, Australian Proteome Analysis Facility, Macquarie University, North Ryde, NSW 2109, Australia Janice L. Joss, Department of Biological Sciences, Macquarie University, North Ryde, NSW 2109, Australia Elizabeth Deane, Department of Biological Sciences, Macquarie University, North Ryde, NSW 2109, Australia Nicolle H. Packer, Biomolecular Frontiers Research Centre, Department of Chemistry and Biomolecular Sciences, Macquarie University, Building E8C Room 307, North Ryde, NSW 2109, Australia Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description: Glycosylation effects on cancer development Content Type Journal Article Pages 1-2 DOI 10.1007/s10719-012-9448-4 Authors Sen-itiroh Hakomori, Division of Biomembrane Research, Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122, USA Richard D. Cummings, Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    We have analyzed the structures of glycosphingolipids and intracellular free glycans in human cancers. In our previous study, trace amounts of free N -acetylneuraminic acid (Neu5Ac)-containing complex-type N -glycans with a single GlcNAc at each reducing terminus (Gn1 type) was found to accumulate intracellularly in colorectal cancers, but were undetectable in most normal colorectal epithelial cells. Here, we used cancer glycomic analyses to reveal that substantial amounts of free Neu5Ac-containing complex-type N -glycans, almost all of which were α2,6-Neu5Ac-linked, accumulated in the pancreatic cancer cells from three out of five patients, but were undetectable in normal pancreatic cells from all five cases. These molecular species were mostly composed of five kinds of glycans having a sequence Neu5Ac-Gal-GlcNAc-Man-Man-GlcNAc and one with the following sequence Neu5Ac-Gal-GlcNAc-Man-(Man-)Man-GlcNAc. The most abundant glycan was Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-3Manβ1-4GlcNAc, followed by Neu5Acα2-6Galβ1-4GlcNAcβ1-2Manα1-6Manβ1-4GlcNAc. This is the first study to show unequivocal evidence for the occurrence of free Neu5Ac-linked N -glycans in human cancer tissues. Our findings suggest that free Neu5Ac-linked glycans may serve as a useful tumor marker. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9435-9 Authors Masahiko Yabu, Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Hiroaki Korekane, Systems Glycobiology Research Group, Chemical Biology Department, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Hidenori Takahashi, Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Hiroaki Ohigashi, Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Osamu Ishikawa, Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Yasuhide Miyamoto, Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka, 537-8511 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Sulfatides, 3- O -sulfogalactosylceramides, are known to have multifunctional properties. These molecules are distributed in various tissues of mammals, where they are synthesized from galactosylceramides by sulfation at C3 of the galactosyl residue. Although this reaction is specifically catalyzed by cerebroside sulfotransferase (CST), the mechanisms underlying the transcriptional regulation of this enzyme are not understood. With respect to this issue, we previously found potential sequences of peroxisome proliferator-activated receptor (PPAR) response element on upstream regions of the mouse CST gene and presumed the possible regulation by the nuclear receptor PPARα. To confirm this hypothesis, we treated wild-type and Ppara -null mice with the specific PPARα agonist fenofibrate and examined the amounts of sulfatides and CST gene expression in various tissues. Fenofibrate treatment increased sulfatides and CST mRNA levels in the kidney, heart, liver, and small intestine in a PPARα-dependent manner. However, these effects of fenofibrate were absent in the brain or colon. Fenofibrate treatment did not affect the mRNA level of arylsulfatase A, which is the key enzyme for catalyzing desulfation of sulfatides, in any of these six tissues. Analyses of the DNA-binding activity and conventional gene expression targets of PPARα has demonstrated that fenofibrate treatment activated PPARα in the kidney, heart, liver, and small intestine but did not affect the brain or colon. These findings suggest that PPARα activation induces CST gene expression and enhances sulfatide synthesis in mice, which suggests that PPARα is a possible transcriptional regulator for the mouse CST gene. Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9454-6 Authors Takero Nakajima, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Yuji Kamijo, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Huang Yuzhe, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Takefumi Kimura, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Naoki Tanaka, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Eiko Sugiyama, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Kozo Nakamura, Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, Minami-Minowa, Kami-Ina, Nagano, Japan Mamoru Kyogashima, Division of Microbiology and Molecular Cell Biology, Nihon Pharmaceutical University, Ina, Kita-Adachi, Saitama, Japan Atsushi Hara, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Toshifumi Aoyama, Department of Metabolic Regulation, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The bisecting GlcNAc is transferred to the core mannose residue of complex or hybrid N-glycans on glycoproteins by the β1,4- N -acetylglucosaminyltransferase III (GlcNAcT-III) or MGAT3. The addition of the bisecting GlcNAc confers unique lectin recognition properties to N-glycans. Thus, LEC10 gain-of-function Chinese hamster ovary (CHO) cells selected for the acquisition of ricin resistance, carry N-glycans with a bisecting GlcNAc, which enhances the binding of the erythroagglutinin E-PHA, but reduces the binding of ricin and galectins-1, -3 and -8. The altered interaction with galactose-binding lectins suggests that the bisecting GlcNAc affects N-glycan conformation. LEC10 mutants expressing polyoma middle T antigen (PyMT) exhibit reduced growth factor signaling. Furthermore, PyMT-induced mammary tumors lacking MGAT3, progress more rapidly than tumors with the bisecting GlcNAc on N-glycans of cell surface glycoproteins. In recent years, evidence for a new paradigm of cell growth control has emerged involving regulation of cell surface residency of growth factor and cytokine receptors via interactions and cross-linking of their branched N-glycans with a lattice of galectin(s). Specific cross-linking of glycoprotein receptors in the lattice regulates their endocytosis, leading to effects on growth factor-induced signaling. This review will describe evidence that the bisecting GlcNAc of N-glycans regulates cellular signaling and tumor progression, apparently through modulating N-glycan/galectin interactions. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9373-6 Authors Hazuki E. Miwa, Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA Yinghui Song, Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA Richard Alvarez, Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA Richard D. Cummings, Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA Pamela Stanley, Department of Cell Biology, Albert Einstein College of Medicine, New York, NY 10461, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The past 25 years have seen significant advances in understanding the diversity and functions of glycoprotein glycans in Drosophila melanogaster . Genetic screens have captured mutations that reveal important biological activities modulated by glycans, including protein folding and trafficking, as well as cell signaling, tissue morphogenesis, fertility, and viability. Many of these glycan functions have parallels in vertebrate development and disease, providing increasing opportunities to dissect pathologic mechanisms using Drosophila genetics. Advances in the sensitivity of structural analytic techniques have allowed the glycan profiles of wild-type and mutant tissues to be assessed, revealing novel glycan structures that may be functionally analogous to vertebrate glycans. This review describes a selected set of recent advances in understanding the functions of N-linked and O-linked (non-glycosaminoglycan) glycoprotein glycans in Drosophila with emphasis on their relatedness to vertebrate organisms. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9442-x Authors Toshihiko Katoh, The Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA Michael Tiemeyer, The Complex Carbohydrate Research Center, University of Georgia, Athens, GA 30602, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In this study, we purified and characterized the β-xylosidase involved in the turnover of plant complex type N -glycans to homogeneity from mature red tomatoes. Purified β-xylosidase (β-Xyl’ase Le-1) gave a single band with molecular masses of 67 kDa on SDS-PAGE under a reducing condition and 60 kDa on gelfiltration, indicating that β-Xyl’ase Le-1 has a monomeric structure in plant cells. The N- terminal amino acid could not be identified owing to a chemical modification. When pyridylaminated (PA-) N- glycans were used as substrates, β-Xyl’ase Le-1 showed optimum activity at about pH 5 at 40 °C, suggesting that the enzyme functions in a rather acidic circumstance such as in the vacuole or cell wall. β-Xyl’ase Le-1 hydrolyzed the β1-2 xylosyl residue from Man 1 Xyl 1 GlcNAc 2 -PA, Man 1 Xyl 1 Fuc 1 GlcNAc 2 -PA, and Man 2 Xyl 1 Fuc 1 GlcNAc 2 -PA, but not that from Man 3 Xyl 1 GlcNAc 2 -PA or Man 3 Xyl 1 Fuc 1 GlcNAc 2 -PA, indicating that the α1-3 arm mannosyl residue exerts significant steric hindrance for the access of β-Xyl’ase Le-1 to the xylosyl residue, whereas the α1-3 fucosyl residue exerts little effect. These results suggest that the release of the β1-2 xylosyl residue by β-Xyl’ase Le-1 occurs at least after the removal the α-1,3-mannosyl residue in the core trimannosyl unit. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9441-y Authors Daisuke Yokouchi, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Natsuko Ono, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Kosuke Nakamura, Kagome Research Institute, Kagome Co., Ltd., 17 Nishitomiyama, Nasushiobara, Tochigi 329-2762, Japan Megumi Maeda, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Yoshinobu Kimura, Department of Biofunctional Chemistry, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka 1-1-1, Okayama, 700-8530 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Many post-translational modifications, including glycosylation, are pivotal for the structural integrity, location and functional activity of glycoproteins. Sub-populations of proteins that are relocated or functionally changed by such modifications can change resting proteins into active ones, mediating specific effector functions, as in the case of monoclonal antibodies. To ensure safe and efficacious drugs it is essential to employ appropriate robust, quantitative analytical strategies that can (i) perform detailed glycan structural analysis, (ii) characterise specific subsets of glycans to assess known critical features of therapeutic activities (iii) rapidly profile glycan pools for at-line monitoring or high level batch to batch screening. Here we focus on these aspects of glycan analysis, showing how state-of-the-art technologies are required at all stages during the production of recombinant glycotherapeutics. These data can provide insights into processing pathways and suggest markers for intervention at critical control points in bioprocessing and also critical decision points in disease and drug monitoring in patients. Importantly, these tools are now enabling the first glycome/genome studies in large populations, allowing the integration of glycomics into other ‘omics platforms in a systems biology context. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9443-9 Authors Tharmala Tharmalingam, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Barbara Adamczyk, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Margaret A. Doherty, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Louise Royle, Ludger Ltd., Culham Science Centre, Oxfordshire, OX14 3EB UK Pauline M. Rudd, NIBRT Glycobiology Laboratory, NIBRT - The National Institute for Bioprocessing Research and Training, Fosters Avenue, Mount Merrion, Blackrock, Co. Dublin, Ireland Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Recently, we demonstrated that the human xylosyltransferase II (XT-II) has enzymatic activity and is able to catalyze the initial and rate-limiting step in the biosynthesis of glycosaminoglycans (GAGs) like chondroitin and dermatan sulfate, as well as heparan sulfate and heparin. Therefore, this enzyme also very likely assumes a crucial regulatory role in the biosynthesis of proteoglycans (PGs). In this study, we identified and characterized for the first time the XYLT2 gene promoter region and transcription factors involved in its regulation. Several binding sites for members of the Sp1 family of transcription factors were identified as being necessary for transcriptional regulation of the XYLT2 gene. This was determined by mithramycin A treatment, electrophoretic mobility shift and supershift assays, as well as numerous site-directed mutagenesis experiments. Different 5′ and 3′ deletion constructs of the predicted GC rich promoter region, which lacks a canonical TATA and CAAT box, revealed that a 177 nts proximal promoter element is sufficient and indispensable to drive the constitutive transcription in full strength in HepG2 hepatoma cells. In addition, we also detected the transcriptional start site using 5′-RACE (rapid amplification of cDNA ends). Our results provide an insight into transcriptional regulation of the XYLT2 gene and may contribute to understanding the manifold GAG-involving processes in health and disease. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9439-5 Authors Benjamin Müller, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Christian Prante, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Cornelius Knabbe, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Knut Kleesiek, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Christian Götting, Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum NRW, Universitätsklinik der Ruhr-Universität Bochum, 32545 Bad Oeynhausen, Germany Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Natural killer gene complex (NKC) encodes a group of proteins with a single C-type lectin-like domain, (CTLD) which can be subdivided several subfamilies according to their structures and expression patterns. The receptors containing the conserved calcium binding sites in the CTLD fold belong to group II of C-type lectin superfamily and are expressed on myeloid cells and non- myeloid cells. The receptors lacking conserved calcium binding sites in the CTLD fold have evolved to bind ligands other than carbohydrates independently on calcium and thereby are named as C-type lectin-like receptors. The C-type lectin-like receptors are previously thought to be exclusively expressed on natural killer (NK) cells and enable NK cells to discriminate self, missing self or altered self. However, some C-type lectin-like receptors are identified in myeloid cells and are intensely investigated, recently. These myeloid C-type lectin-like receptors, especially Dectin-1 cluster, have a wide variety of ligands, including those of exogenous origin, and play important roles in the physiological functions and pathological processes including immune homeostasis, immune defenses, and immune surveillance. In this review, we summarize each member of the Dectin-1 cluster, including their structural profiles, expression patterns, signaling properties as well as known physiological functions. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9419-9 Authors Jianhui Xie, Key Laboratory of Glycoconjugate Research, Ministry of Health, Department of Biochemistry and Molecular Biology, Shanghai Medical College, Fudan University, Shanghai, 200032 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Human LOX-1/OLR 1 plays a key role in atherogenesis and endothelial dysfunction. The N-glycosylation of LOX-1 has been shown to affect its biological functions in vivo and modulate the pathogenesis of atherosclerosis. However, the N-glycosylation pattern of LOX-1 has not been described yet. The present study was aimed at elucidating the N-glycosylation of recombinant human LOX-1 with regard to N-glycan profile and N-glycosylation sites. Here, an approach using nonspecific protease (Pronase E) digestion followed by MALDI-QIT-TOF MS and multistage MS (MS 3 ) analysis is explored to obtain site-specific N-glycosylation information of recombinant human LOX-1, in combination with glycan structure confirmation through characterizing released glycans using tandem MS. The results reveal that N-glycans structures as well as their corresponding attached site of LOX-1 can be identified simultaneously by direct MS analysis of glycopeptides from non-specific protease digestion. With this approach, one potential glycosylation site of recombinant human LOX-1 on Asn 139 is readily identified and found to carry heterogeneous complex type N-glycans. In addition, manual annotation of multistage MS data utilizing diagnostic ions, which were found to be particularly useful in defining the structure of glycopeptides and glycans was addressed for proper spectra interpretation. The findings described herein will shed new light on further research of the structure-function relationships of LOX-1 N-glycan. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9408-z Authors Yifan Qian, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Xingwang Zhang, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Lei Zhou, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Xiaojing Yun, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Jianhui Xie, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Jiejie Xu, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Yuanyuan Ruan, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Shifang Ren, Key Laboratory of Glycoconjugate Research Ministry of Public Health, Shanghai Medical College, Fudan University, Shanghai, 200032 People’s Republic of China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Rheumatoid arthritis (RA) is an inflammatory disorder that is characterized by persistent recurrence of joint inflammation leading to cartilage and bone destruction. The present anti-arthritis therapies failed to achieve satisfactory remission in all patients; therefore, it is still necessary to develop novel approaches to fulfill the demand in clinic. Here, we reported the therapeutic effects of lactosyl derivative Gu-4, a synthetic compound that was previously identified as a selective inhibitor against leukocyte integrin CD11b, in a bovine type II collagen induced arthritis (CIA) rat model. First, prophylactic administration of Gu-4 (1.2728 mg/kg) to rats by intraperitoneal injection every 2 days from the first day of collagen immunization significantly decreased the incidence of CIA, diminished the mean paw volume increase, and reduced the number of swollen paws. Second, administration of Gu-4 (1.2728 mg/kg) to rats at early-onset stage of CIA prevented the progression of the pathological process of RA, accelerated the remission of paw edema, and declined the arthritis score; after 5 weeks treatment, X-ray and histological examinations were carried out, the ankle joint of hind limb of Gu-4 treated CIA rats exhibited slighter bone erosion and much less inflammatory cell infiltration compared to those of saline treated animals; furthermore, Gu-4 remarkably attenuated the production of rheumatoid factor (RF) in the serum of CIA rats as determined by ELISA. Moreover, we performed in vitro lymphocyte proliferation assay and found that Gu-4 significantly inhibited the proliferation of splenic lymphocytes isolated from CIA rats in a dose-dependent manner. Our results suggest that Gu-4 can effectively ameliorate CIA and might be an alternative option for the treatment of RA. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9407-0 Authors Jie Fan, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Huiting Zhou, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Shihui Wang, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Hailian Wang, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Yushun Zhang, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Yingtao Guo, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Qing Li, State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, 100083 China Zhongjun Li, State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, 100083 China Zhihui Zhao, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing, 210046 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Despite recent technical advances in glycan analysis, the rapidly growing field of glycomics still lacks methods that are high throughput and robust, and yet allow detailed and reliable identification of different glycans. LC-MS-MS 2 methods have a large potential for glycan analysis as they enable separation and identification of different glycans, including structural isomers. The major drawback is the complexity of the data with different charge states and adduct combinations. In practice, manual data analysis, still largely used for MALDI-TOF data, is no more achievable for LC-MS-MS 2 data. To solve the problem, we developed a glycan analysis software GlycanID for the analysis of LC-MS-MS 2 data to identify and profile glycan compositions in combination with existing proteomic software. IgG was used as an example of an individual glycoprotein and extracted cell surface proteins of human fibroblasts as a more complex sample to demonstrate the power of the novel data analysis approach. N-glycans were isolated from the samples and analyzed as permethylated sugar alditols by LC-MS-MS 2 , permitting semiquantitative glycan profiling. The data analysis consisted of five steps: 1) extraction of LC-MS features and MS 2 spectra, 2) mapping potential glycans based on feature distribution, 3) matching the feature masses with a glycan composition database and de novo generated compositions, 4) scoring MS 2 spectra with theoretical glycan fragments, and 5) composing the glycan profile for the identified glycan compositions. The resulting N-glycan profile of IgG revealed 28 glycan compositions and was in good correlation with the published IgG profile. More than 50 glycan compositions were reliably identified from the cell surface N-glycan profile of human fibroblasts. Use of the GlycanID software made relatively rapid analysis of complex glycan LC-MS-MS 2 data feasible. The results demonstrate that the complexity of glycan LC-MS-MS 2 data can be used as an asset to increase the reliability of the identifications. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9412-3 Authors Hannu Peltoniemi, Applied Numerics Ltd, Nuottapolku 10 A 8, 00330 Helsinki, Finland Suvi Natunen, Finnish Red Cross Blood Service, Helsinki, Finland Ilja Ritamo, Finnish Red Cross Blood Service, Helsinki, Finland Leena Valmu, Finnish Red Cross Blood Service, Helsinki, Finland Jarkko Räbinä, Finnish Red Cross Blood Service, Helsinki, Finland Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Proteoglycans have been studied to a limited extent in lymphoid cells. In this study we have investigated the expression of proteoglycans in B-cells, CD4+ T-cells, CD8+ T-cells, natural killer cells, as well as in nine different cell lines established from patients with lymphoid malignancies. Serglycin was the major proteoglycan expressed at mRNA level by the primary lymphocytes. None of the syndecans or glycpicans was detected at mRNA level in the primary lymphocytes, except for syndecan-4 in CD4+ T-cells and CD8+ T-cells. All lymphoid cell lines expressed serglycin mRNA, as well as one or several members of the syndecan and glypican families. Further, increased synthesis of proteoglycans was found in the cell lines compared to the primary lymphocytes, as well as the presence of heparan sulfate on the cell surface of five of the cells lines. Western blot analysis showed a close correlation between serglycin mRNA level and expression of serglycin core protein. Our results show that serglycin is a major proteoglycan in all the normal lymphoid cells and that these cells carry little, or none, proteoglycans on the cell surface. Serglycin was also a major proteoglycan in the malignant lymphoid cells, but these also expressed one or more types of cell surface proteoglycans. Thus, malignant transformation of lymphoid cells may be followed by increased synthesis of proteoglycans and expression of cell surface proteoglycans. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9427-9 Authors Bodil Fadnes, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Anne Husebekk, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Gunbjørg Svineng, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Øystein Rekdal, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Masaki Yanagishita, Tokyo Medical and Dental University, Tokyo, Japan Svein O. Kolset, Department of Nutrition, University of Oslo, Oslo, Norway Lars Uhlin-Hansen, Institute of Medical Biology, Faculty of Health Sciences, University of Tromsø, Tromsø, Norway Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Mucin-type O -linked glycoproteins are known for regulating many aspects of cell activity but remains a challenge to detect under physiological conditions which is due to the diversity of O -glycosylation and the lack of universal method. Here a direct labeling strategy for in situ visualizing of mucin-type O -linked glycoproteins on living cells has been developed. The strategy utilizes the combination of metabolic engineering and chemical probing technologies. Treating cells with an unnatural sugar, 2-keto Ac 4 GalNAc analogue (2-keto isostere of GalNAc) to generate keto groups upon cells, followed by chemoselective ligation of keto groups on cells with a fluorescent tag, fluorescein-5-thiosemicarbazide (FTSC), provides a promising platform to probing mucin-type O -glycosylation on living cells. The FTSC conjugates illustrated very similar fluorescent spectra as FITC, a fluorescent tag widely used in proteomics, indicating good compatibility with commonly used fluorescent equipments. The established method eliminated the need of an additional fluorescent amplification step. Cells after being treated with the method maintained a rather high level of viability of 84.3 %. Finally, the assay has been successfully applied to image the expression of mucin-type O -linked glycoproteins within CHO and HeLa cells. Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9425-y Authors Ying Zhang, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Yujiao Sun, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Zhongfu Wang, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Linjuan Huang, Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University, Xi’an, 710069 People’s Republic of China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Glycoconjugates (GCs) are recognized as stimulation and signaling agents, affecting cell adhesion, activation, and growth of living organisms. Among GC targets, macrophages are considered ideal since they play a central role in inflammation and immune responses against foreign agents. In this context, we studied the effects of highly selective GCs in neutralizing toxin factors produced by B. anthracis during phagocytosis using murine macrophages. The effects of GCs were studied under three conditions: A) prior to , B) during , and C) following exposure of macrophages to B. anthracis individual toxin (protective antigen [PA], edema factor [EF], lethal factor [LF] or toxin complexes (PA-EF-LF, PA-EF, and PA-LF). We employed ex vivo phagocytosis and post-phagocytosis analysis including direct microscopic observation of macrophage viability, and macrophage activation. Our results demonstrated that macrophages are more prone to adhere to GC-altered PA-EF-LF, PA-EF, and PA-LF toxin complexes. This adhesion results in a higher phagocytosis rate and toxin complex neutralization during phagocytosis. In addition, GCs enhance macrophage viability, activate macrophages, and stimulate nitric oxide (NO) production. The present study may be helpful in identifying GC ligands with toxin-neutralizing and/or immunomodulating properties. In addition, our study could suggest GCs as new targets for existing vaccines and the prospective development of vaccines and immunomodulators used to combat the effects of B. anthracis . Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9446-6 Authors Olga Tarasenko, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Ashley Scott, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA April Jones, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Lee Soderberg, Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR, USA Pierre Alusta, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In the past decade, the identification of most genes involved in Congenital Disorders of Glycosylation (CDG) (type I) was achieved by a combination of biochemical, cell biological and glycobiological investigations. This has been truly successful for CDG-I, because the candidate genes could be selected on the basis of the homology of the synthetic pathway of the dolichol linked oligosaccharide in human and yeast. On the contrary, only a few CDG-II defects were elucidated, be it that some of the discoveries represent wonderful breakthroughs, like e.g , the identification of the COG defects. In general, many rare genetic defects have been identified by positional cloning. However, only a few types of CDG have effectively been elucidated by linkage analysis and so-called reverse genetics. The reason is that the families were relatively small and could—except for CDG-PMM2—not be pooled for analysis. Hence, a large number of CDG cases has long remained unsolved because the search for the culprit gene was very laborious, due to the heterogeneous phenotype and the myriad of candidate defects. This has changed when homozygosity mapping came of age, because it could be applied to small (consanguineous) families. Many novel CDG genes have been discovered in this way. But the best has yet to come: what we are currently witnessing, is an explosion of novel CDG defects, thanks to exome sequencing: seven novel types were published over a period of only two years. It is expected that exome sequencing will soon become a diagnostic tool, that will continuously uncover new facets of this fascinating group of diseases. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9445-7 Authors Gert Matthijs, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Daisy Rymen, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium María Beatriz Bistué Millón, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Erika Souche, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Valérie Race, Center for Human Genetics, University of Leuven, Herestraat 49, 3000 Leuven, Belgium Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    This review summarizes the analytical advances made during the last several years in the structural and quantitative determinations of glycoproteins in complex biological mixtures. The main analytical techniques used in the fields of glycomics and glycoproteomics involve different modes of mass spectrometry and their combinations with capillary separation methods such as microcolumn liquid chromatography and capillary electrophoresis. The need for high-sensitivity measurements have been emphasized in the oligosaccharide profiling used in the field of biomarker discovery through MALDI mass spectrometry. High-sensitivity profiling of both glycans and glycopeptides from biological fluids and tissue extracts has been aided significantly through lectin preconcentration and the uses of affinity chromatography. Content Type Journal Article Pages 1-29 DOI 10.1007/s10719-012-9444-8 Authors Milos V. Novotny, Department of Chemistry, Indiana University, Bloomington, IN, USA William R. Alley Jr., Department of Chemistry, Indiana University, Bloomington, IN, USA Benjamin F. Mann, Department of Chemistry, Indiana University, Bloomington, IN, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Inborn errors in glycoconjugate biosynthesis termed ‘Congenital Disorders of Glycosylation’ (CDG) comprise a rapidly expanding group of metabolic diseases in man. Up till now more than 60 different inherited disorders in N- and O-glycosylation pathways have been identified. They affect the biosynthesis of glycan moieties linked to proteins as well as lipids. Due to failures in protein glycosylation, CDG patients suffer from multi systemic disorders, which mostly present with severe psychomotor and mental retardations, muscular impairment, ataxia, failure to thrive and developmental delay. Although improved biochemical and genetic investigations led to identification of a variety of new molecular defects in glycoconjugate biosynthesis, effective therapies for most types of the CDG are so far not available. Therefore, intensive investigations on treatment options for this group of diseases have been carried out in recent years. Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9447-5 Authors Christian Thiel, Center for Child and Adolescent Medicine, Center for Metabolic Diseases Heidelberg, Kinderheilkunde I Im Neuenheimer Feld 433, 69120 Heidelberg, Germany Christian Körner, Center for Child and Adolescent Medicine, Center for Metabolic Diseases Heidelberg, Kinderheilkunde I Im Neuenheimer Feld 433, 69120 Heidelberg, Germany Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Despite numerous original publications describing the structural complexity of N- and O-linked glycans on glycoproteins, only very few answer the basic question of which particular glycans are linked to which amino acid residues along the polypeptide chain. Such structural information is of fundamental importance for understanding the biological roles of complex glycosylations as well as deciphering their non-template driven biosynthesis. This review focuses on presenting and commenting on recent strategies, specifically aimed at identifying the glycoproteome of cultured cells and biological samples, using targeted and global enrichment procedures and utilizing the high resolution power, high through-put capacity and complementary fragmentation techniques of tandem mass spectrometry. The goal is to give an update of this emerging field of protein and glyco-sciences and suggest routes to bridge the data gap between the two aspects of glycoprotein characteristics, i.e. glycan structures and their attachment sites. Content Type Journal Article Pages 1-18 DOI 10.1007/s10719-012-9438-6 Authors Jonas Nilsson, Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at the University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, 413 45 Sweden Adnan Halim, Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at the University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, 413 45 Sweden Ammi Grahn, Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at the University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, 413 45 Sweden Göran Larson, Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, The Sahlgrenska Academy at the University of Gothenburg, Sahlgrenska University Hospital, Gothenburg, 413 45 Sweden Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Among the “omics”, glycomics is one of the most complex fields and needs complementary strategies of analysis to decipher the “glycan dictionary”. As an alternative method, which has developed since the beginning of the 21st century, lectin array technology could generate relevant information related to glycan motifs, accessibility and a number of other valuable insights from molecules (purified and non-purified) or cells. Based on a cell line model, this study deals with the key parameters that influence the whole cell surface glycan interaction with lectin arrays and the consequences on the interpretation and reliability of the results. The comparison between the adherent and suspension forms of Chinese Hamster Ovary (CHO) cells, showed respective glycan signatures, which could be inhibited specifically by neoglycoproteins. The modifications of the respective glycan signatures were also revealed according to the detachment modes and cell growth conditions. Finally the power of lectin array technology was highlighted by the possibility of selecting and characterizing a specific clone from the mother cell line, based on the slight difference determination in the respective glycan signatures. Content Type Journal Article Category Glycoarray Section Pages 1-9 DOI 10.1007/s10719-012-9433-y Authors Ludovic Landemarre, GLYcoDIAG, Université d’Orléans, 45067 Orléans cedex 2, France Perrine Cancellieri, GLYcoDIAG, Université d’Orléans, 45067 Orléans cedex 2, France Eric Duverger, GlycoBiochimie/Département de Biologie, Université d’Orléans, 45067 Orléans cedex 2, France Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Brine shrimp are primitive crustacean arthropodal model organisms, second to daphnia, which can survive in high-salinity environments. Their oviposited cysts, cuticle-covered diapausing eggs, are highly resistant to dryness. To elucidate specialties of brine shrimp, this study characterized glycosphingolipids, which are signal transduction-associated material. A group of novel and complex fucosyl glycosphingolipids were separated and identified from cysts of the brine shrimp Artemia franciscana by repeated lipid extraction, alkaline methanolysis, acid treatment, successive column chromatography, and post-source decay measurements by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Structures of the glycosphingolipids were elucidated by conventional structural characterization and mass spectrometry, and the compounds were identified as GlcNAcβ1-3GalNAcβ1-4(GlcNAcα1-2Fucα1-3)GlcNAcβ1-3Manβ1-4Glcβ1-Cer, GalNAcβ1-4(Fucα1-3)GlcNAcβ1-3GalNAcβ1-4(GlcNAcα1-2Fucα1-3)GlcNAcβ1-3Manβ1-4Glcβ1-Cer, and GalNAcβ1-4(GlcNAcα1-2Fucα1-3)GlcNAcβ1-3GalNAcβ1-4(GlcNAcα1-2Fucα1-3)GlcNAcβ1-3Manβ1-4Glcβ1-Cer. These compounds also contained a branching, non-arthro-series disaccharide with an α-GlcNAc terminus, similar to that found in a previously reported ceramide hexasaccharide (III 3 (GlcNAcα2Fucα)-At 4 Cer). The glycans within these complex GSLs are longer than reported glycans of the animal kingdom containing α-GlcNAc terminus. These complex GSLs as well as the longest GSL with ten sugar residues, ceramide decasaccharide (CDeS), contain the fucosylated LacdiNAc sequence reported to associate with parasitism/immunosuppression and the α-GlcNAc terminus reported to show a certain antibacterial effect in other reports. CDeS, the longest GSL of this species, was found in the highest amount, which indicates that CDeS may be functionally important. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9436-8 Authors Hisao Kojima, Institute of Glycoscience, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan Yukako Tohsato, Department of Bioinformatics, College of Life Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan Kazuya Kabayama, Institute of Glycoscience, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan Saki Itonori, Department of Chemistry, Faculty of Liberal Arts and Education, Shiga University, 2-5-1 Hiratsu, Otsu, Shiga 520-0862, Japan Masahiro Ito, Department of Bioinformatics, College of Life Sciences, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Galanthus nivalis agglutinin (GNA)-related lectin family, a superfamily of strictly mannose-binding specific lectins widespread among monocotyledonous plants, is well-known to possess a broad range of biological functions such as anti-tumor, anti-viral and anti-fungal activities. Herein, we mainly focused on exploring the precise molecular mechanisms by which GNA-related lectins induce cancer cell apoptotic and autophagic death targeting mitochondria-mediated ROS-p38-p53 apoptotic or autophagic pathway, Ras-Raf and PI3K-Akt anti-apoptotic or anti-autophagic pathways. In addition, we further discussed the molecular mechanisms of GNA-related lectins exerting anti-viral activities by blocking the entry of the virus into its target cells, preventing transmission of the virus as well as forcing virus to delete glycan in its envelope protein and triggering neutralizing antibody. In conclusion, these findings may provide a new perspective of GNA-related lectins as potential drugs for cancer and virus therapeutics in the future. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9440-z Authors Lei Wu, School of Life Sciences and Key Laboratory of Bio-resources and Eco-environment, Sichuan University, Ministry of Education, Chengdu, 610064 China Jin-ku Bao, School of Life Sciences and Key Laboratory of Bio-resources and Eco-environment, Sichuan University, Ministry of Education, Chengdu, 610064 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    A new mannose-recognizing lectin (MOL) was purified on an asialofetuin-column from fruiting bodies of Marasmius oreades grown in Japan. The lectin (MOA) from the fruiting bodies of the same fungi is well known to be a ribosome-inactivating type lectin that recognizes blood-group B sugar. However, in our preliminary investigation, MOA was not found in Japanese fruiting bodies of M. oreades , and instead, MOL was isolated. Gel filtration showed MOL is a homodimer noncovalently associated with two subunits of 13 kDa. The N-terminal sequence of MOL was blocked. The sequence of MOL was determined by cloning from cDNA and by protein sequencing of enzyme-digested peptides. The sequence shows mannose-binding motifs of bulb-type mannose-binding lectins from plants, and similarity to the sequences. Analyses of sugar-binding specificity by hemagglutination inhibition revealed the preference of MOL toward mannose and thyroglobulin, but asialofetuin was the strongest inhibitor of glycoproteins tested. Furthermore, glycan-array analysis showed that the specificity pattern of MOL was different from those of typical mannose-specific lectins. MOL preferred complex–type N-glycans rather than high-mannose N-glycans. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9401-6 Authors Michiko Shimokawa, Department of Applied Biological Chemistry, The United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065 Japan Ayako Fukudome, Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065 Japan Ryoko Yamashita, Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065 Japan Yuji Minami, Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065 Japan Fumio Yagi, Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065 Japan Hiroaki Tateno, Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Central 2, 1-1-1 Umezono, Ibaraki, 305-8568 Japan Jun Hirabayashi, Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Central 2, 1-1-1 Umezono, Ibaraki, 305-8568 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Rice ( Oryza sativa ) expresses different putative carbohydrate-binding proteins belonging to the class of lectins containing an Euonymus lectin (EUL)-related domain, one of them being OrysaEULS2. The OrysaEULS2 sequence consists of a 56 amino acid N-terminal domain followed by the EUL sequence. In this paper the original sequence of the EUL domain of OrysaEULS2 and some mutant forms have been expressed in Pichia pastoris . Subsequently, the recombinant proteins were purified and their carbohydrate binding properties determined. Analysis of the original protein on the glycan array revealed interaction with mannose containing structures and to a lesser extent with glycans containing lactosamine related structures. It was shown that mutation of tryptophan residue 134 into leucine resulted in an almost complete loss of carbohydrate binding activity of OrysaEULS2. Our results show that the EUL domain in OrysaEULS2 interacts with glycan structures, and hence can be considered as a lectin. However, the binding of the protein with the array is much weaker than that of other EUL-related lectins. Furthermore, our results indicate that gene divergence within the family of EUL-related lectins lead to changes in carbohydrate binding specificity. Content Type Journal Article Pages 1-13 DOI 10.1007/s10719-012-9405-2 Authors Bassam Al Atalah, Laboratory of Biochemistry and Glycobiology, Department of Molecular Biotechnology, Ghent University, Coupure links 653, 9000 Ghent, Belgium Pierre Rougé, Signaux et Messages Cellulaires chez les Végétaux, UMR CNRS-UPS 5546, Pole de Biotechnologie végétale, BP 17, 24 Chemin de Borde Rouge, Castanet-Tolosan, 31326 France David F. Smith, Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA Paul Proost, Laboratory of Molecular Immunology, Rega Institute for Medical Research, Katholieke Universiteit Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium Yi Lasanajak, Department of Biochemistry, Emory University School of Medicine, Atlanta, GA 30322, USA Els J. M. Van Damme, Laboratory of Biochemistry and Glycobiology, Department of Molecular Biotechnology, Ghent University, Coupure links 653, 9000 Ghent, Belgium Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Playing an important role in a broad range of biological and pathological processes, sialylation has been drawing wide interest. The efficient sialoglycopeptides enrichment methods are therefore attracting considerable attention. In this paper, we first compared two conventional enrichment methods, lectin and TiO 2 , and analyzed their characteristics. Furthermore, considering the highly negatively charged nature of sialic acids, we developed a new strategy, peptide immobilized pH gradient isoelectric focusing (IPG-IEF) assisted TiO 2 chromatography (PIAT), for the highly efficient enrichment of sialoglycopeptides. In this method, peptides were first separated into 24 fractions using peptide IPG-IEF. Sialoglycopeptides were relatively concentrated in low-pH fractions of the immobilized pH strips and were captured using TiO 2 chromatography. As a result, 614 N-glycosylation sites were identified in 582 sialoglycopeptides within 322 sialoglycoproteins from rat liver using PIAT. To our knowledge, this work represents one of the most comprehensive sialoglycoproteomic analyses in general and exhibits the largest database of sialoglycoproteome in rat liver currently. So the new strategy introduced here exhibits high efficiency and universality in the sialoglycopeptide enrichment, and is a powerful tool for sialoglycoproteome exploration. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9404-3 Authors Weiqian Cao, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Jing Cao, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Jiangming Huang, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Lei Zhang, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Jun Yao, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Haoqi Xu, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Pengyuan Yang, Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai, 200433 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Cell surface glycoproteins are one of the most frequently observed phenomena correlated with malignant growth. Hepatocellular carcinoma (HCC) is one of the most malignant tumors in the world. The majority of hepatocellular carcinoma cell surface proteins are modified by glycosylation in the process of tumor invasion and metastasis. Therefore, characterization of cell surface glycoproteins can provide important information for diagnosis and treatment of liver cancer, and also represent a promising source of potential diagnostic biomarkers and therapeutic targets for hepatocellular carcinoma. However, cell surface glycoproteins of HCC have been seldom identified by proteomics approaches because of their hydrophobic nature, poor solubility, and low abundance. The recently developed cell surface-capturing (CSC) technique was an approach specifically targeted at membrane glycoproteins involving the affinity capture of membrane glycoproteins using glycan biotinylation labeling on intact cell surfaces. To characterize the cell surface glycoproteome and probe the mechanism of tumor invasion and metastasis of HCC, we have modified and evaluated the cell surface-capturing strategy, and applied it for surface glycoproteomic analysis of hepatocellular carcinoma cells. In total, 119 glycosylation sites on 116 unique glycopeptides were identified, corresponding to 79 different protein species. Of these, 65 (54.6 %) new predicted glycosylation sites were identified that had not previously been determined experimentally. Among the identified glycoproteins, 82 % were classified as membrane proteins by a database search, 68 % had transmembrane domains (TMDs), and 24 % were predicted to contain 2–13 TMDs. Moreover, a total of 26 CD antigens with 50 glycopeptides were detected in the membrane glycoproteins of hepatocellular carcinoma cells, comprising 43 % of the total glycopeptides identified. Many of these identified glycoproteins are associated with cancer such as CD44, CD147 and EGFR. This is a systematic characterization of cell surface glycoproteins of HCC. The membrane glycoproteins identified in this study provide very useful information for probing the mechanism of liver cancer invasion and metastasis. Content Type Journal Article Pages 1-14 DOI 10.1007/s10719-012-9420-3 Authors Wei Mi, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Wei Jia, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Zhaobin Zheng, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Jinglan Wang, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Yun Cai, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Wantao Ying, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Xiaohong Qian, State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, 33 Life Park Road, Changping District, Beijing 102206, People’s Republic of China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Sialic acid-containing glycosphingolipids, gangliosides are highly expressed in human cancer cells and regulate cell signals transduced via membrane microdomains. Generally, disialyl gangliosides enhance tumor phenotypes, while monosialyl gangliosides suppress them. In particular, gangliosides GD3 and GD2 are highly expressed in melanomas and small cell lung cancer cells, and their expression cause increased cell growth and invasion. In osteosarcomas, expression of GD3 and GD2 also enhanced cell invasion and motility, and caused increased phosphorylation of focal adhesion kinase and paxillin. In addition to focal adhesion kinase, Lyn kinase was also activated by GD3/GD2 expression, leading to the phosphorylation of paxillin. In contrast with melanoma cells, osteosarcomas showed reduced cell adhesion with increased phosphorylation of paxillin. Thus, increased expression of GD3/GD2 caused enhanced activation of signaling molecules, leading to distinct phenotypes between melanomas and osteosarcomas, i.e. increased and decreased adhesion activity. Thus, whole features of glycolipid-enriched microdomain/rafts formed in the individual cancer types seem to determine the main signaling pathway and biological outcome. Content Type Journal Article Pages 1-6 DOI 10.1007/s10719-012-9423-0 Authors Koichi Furukawa, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya, 466-0065 Japan Kazunori Hamamura, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya, 466-0065 Japan Yuki Ohkawa, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya, 466-0065 Japan Yuhsuke Ohmi, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya, 466-0065 Japan Keiko Furukawa, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya, 466-0065 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    A water-soluble polysaccharide CSPS-2B-2 with a molecular mass of 8.8 kDa, was obtained from the fruits of Capparis spinosa L. Chemical and NMR spectral analysis verified CSPS-2B-2 was a linear poly-(1-4)-α-D-galactopyranosyluronic acid in which 12.9 ± 0.4 % of carboxyl groups existed as methyl ester and 2.6 ± 0.1 % of D-Gal p A residues were acetylated. A sulfated derivative Sul-2B-2 with a sulfation degree of 0.88 ± 0.02 was prepared via the substitution of C-2 and/or C-3 of Gal p A residues in CSPS-2B-2. Bioassay on the complement and coagulation system demonstrated that Sul-2B-2 (CH 50 : 3.5 ± 0.2 μg/mL) had a stronger inhibitory effect on the activation of complement system through the classic pathway than that of heparin (CH 50 : 8.9 ± 0.3 μg/mL). Interestingly, Sul-2B-2 at low dose even middle dose (for example 52 μg/mL) had no effect on coagulation system, which was totally different from heparin. Thus, our observation indicated that Sul-2B-2 was more efficient than heparin in inhibiting the activation of the complement system through classical pathway and exhibiting a relatively less anti-coagulant activity. These results suggested that the sulfated derivative Sul-2B-2 prepared from the homogalacturonan in the fruits of Capparis spinosa L , might be a promising drug candidate in case of necessary therapeutic complement inhibition. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9418-x Authors Huijun Wang, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203 China Hongwei Wang, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203 China Songshan Shi, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203 China Jinyou Duan, College of Science, Northwest A&F University, Yangling, 712100 Shaanxi, China Shunchun Wang, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Glycosylation is an important method for the structural modification of various flavonols, resulting in the glycosides with increased solubility, stability and bioavailability compared with the corresponding aglycone. From the physiological point of view, glycosylation of plant flavonoids is of importance and interest. However, it is notoriously complicated that flavonols such as quercetin, kaempferol and myricetin, are glucosylated regioselectively at the specific position by chemical method. Compared to the chemical method, enzymatic synthesis present several advantages, such as mild reaction condition, high stereo or region selectivity, no protection/deprotection and high yield. UGT78D1 is a flavonol-specific glycosyltransferase, responsible for transferring rhamnose or glucose to the 3-OH position in vitro . In this study, the activity of UGT78D1 was tested against 28 flavonoids acceptors using UDP-glucose as donor nucleoside in vitro , and 5 acceptors, quercetin, myricetin, kaempferol, fisetin and isorhamnetin, were discovered to be glucosylated at 3-OH position. Herein, the small-scale 3-O-glucosylated quercetin, kaempferol and myricetin were synthesized by UGT78D1 and their chemical structures were confirmed by 1 H and 13 C nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HRMS). Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9410-5 Authors Guangxiang Ren, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Jingli Hou, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Qinghong Fang, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Hong Sun, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Xiaoyan Liu, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Lianwen Zhang, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Peng George Wang, College of Pharmacy and State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071 People’s Republic of China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In the majority of congenital disorders of glycosylation, the assembly of the glycan precursor GlcNAc 2 Man 9 Glc 3 on the polyprenol carrier dolichyl-pyrophosphate is compromised. Because N-linked glycosylation is essential to life, most types of congenital disorders of glycosylation represent partial losses of enzymatic activity. Consequently, increased availability of substrates along the glycosylation pathway can be beneficial to increase product formation by the compromised enzymes. Recently, we showed that increased dolichol availability and improved N-linked glycosylation can be achieved by inhibition of squalene biosynthesis. This review summarizes the current knowledge on the biosynthesis of dolichol-linked glycans with respect to deficiencies in N-linked glycosylation. Additionally, perspectives on therapeutic treatments targeting dolichol and dolichol-linked glycan biosynthesis are examined. Content Type Journal Article Pages 1-6 DOI 10.1007/s10719-012-9417-y Authors Michael Welti, Institute of Physiology, University of Zürich, Winterthurerstrasse 190, 8057 Zürich, Switzerland Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Protein glycosylation is acknowledged as one of the major posttranslational modifications that elicit significant effects on protein folding, conformation, distribution, stability, and activity. The changes in glycoprotein abundance, glycosylation degree, and glycan structure are associated with a variety of diseases. Therefore, the quantitative study of glycoproteomics has become a new and popular research topic, and is quickly emerging as an important technique for biomarker discovery. Mass spectrometry-based protein quantification technologies provide a powerful tool for the systematic and quantitative assessment of the quantitative differences in the protein profiles of different samples. Combined with various glycoprotein/glycopeptide enrichment strategies and other glycoprotein analysis methods, these techniques have been further developed for application in quantitative glycoproteomics. A comprehensive quantitative analysis of the glycoproteome in a complex biological sample remains challenging because of the enormous complexity of biological samples, intrinsic characteristics of glycoproteins, and lack of universal quantitative technology. In this review, recently developed technologies in quantitative glycoproteome, especially those focused on two of the most common types of glycosylation (N-linked and O-linked glycoproteome), were summarized. The strengths and weaknesses of the various approaches were also discussed. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9398-x Authors Ying Zhang, Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032 China Hongrui Yin, Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032 China Haojie Lu, Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The α-1,3-glucosyltransferase WaaG is involved in the synthesis of the core region of lipopolysaccharides in E. coli . A fragment-based screening for inhibitors of the WaaG glycosyltrasferase donor site has been performed using NMR spectroscopy. Docking simulations were performed for three of the compounds of the fragment library that had shown binding activity towards WaaG and yielded 3D models for the respective complexes. The three ligands share a hetero-bicyclic ring system as a common structural motif and they compete with UDP-Glc for binding. Interestingly, one of the compounds promoted binding of uridine to WaaG, as seen from STD NMR titrations, suggesting a different binding mode for this ligand. We propose these compounds as scaffolds for the design of selective high-affinity inhibitors of WaaG. Binding of natural substrates, enzymatic activity and donor substrate selectivity were also investigated by NMR spectroscopy. Molecular dynamics simulations of WaaG were carried out with and without bound UDP and revealed structural changes compared to the crystal structure and also variations in flexibility for some amino acid residues between the two WaaG systems studied. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9411-4 Authors Jens Landström, Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91 Stockholm, Sweden Karina Persson, Department of Chemistry, Umeå University, 901 87 Umeå, Sweden Christoph Rademacher, Institute of Chemistry, University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck, Germany Magnus Lundborg, Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91 Stockholm, Sweden Warren Wakarchuk, National Research Council of Canada, Institute for Biological Sciences, 100 Sussex Drive, Ottawa, ON K1A 0R6, Canada Thomas Peters, Institute of Chemistry, University of Luebeck, Ratzeburger Allee 160, 23538 Luebeck, Germany Göran Widmalm, Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91 Stockholm, Sweden Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    C-type lectin-like receptor 2 (CLEC-2) is a newly identified type II transmembrane protein belonging to the C-type lectin family molecules, which acts as a cell-surface receptor for snake venom toxin rhodocytin and tumor antigen podoplanin. We previously demonstrated that the full-length mouse CLEC-2 (mCLEC-2) can be cleaved into soluble form. Elevated levels of soluble forms of membrane proteins in circulating blood may reflect increased expression of membrane proteins and disease activities. In the present study, we clarified the domain and sites contributing to the production of soluble mCLEC-2. The shedding process can be positively regulated by protein kinase C (PKC). Moreover, we explored the possibility that human CLEC-2 (hCLEC-2) may also be proteolyticly cleaved and released as a soluble form. We have observed that the production of soluble hCLEC-2 could be induced by phorbol ester (PMA) in cells stably transfected with hCLEC-2 cDNA. Further studies may explore therapeutic and diagnostic applications of soluble hCLEC-2 in platelet-related diseases. Content Type Journal Article Pages 1-7 DOI 10.1007/s10719-012-9413-2 Authors Min Fei, Jiangsu Institute of Hematology, Soochow University, Suzhou, Jiangsu, China Lei Zhou, Department of Biochemistry, Shanghai Medical School, Fudan University, Shanghai, 200032 People’s Republic of China Jianhui Xie, Department of Biochemistry, Shanghai Medical School, Fudan University, Shanghai, 200032 People’s Republic of China Yuanyuan Ruan, Department of Biochemistry, Shanghai Medical School, Fudan University, Shanghai, 200032 People’s Republic of China Jiejie Xu, Department of Biochemistry, Shanghai Medical School, Fudan University, Shanghai, 200032 People’s Republic of China Songbin He, Department of General Surgery, The First Affiliated Hospital of Soochow University, Suzhou, China Hongjie Shen, Jiangsu Institute of Hematology, Soochow University, Suzhou, Jiangsu, China Yumin Hu, Institute of Medical Biotechnology, Soochow University, Suzhou, China Shifang Ren, Department of Biochemistry, Shanghai Medical School, Fudan University, Shanghai, 200032 People’s Republic of China Changgeng Ruan, Jiangsu Institute of Hematology, Soochow University, Suzhou, Jiangsu, China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    N -acetylglucosaminyltransferase (GnT)-IV a is a key enzyme that catalyzes the formation of the GlcNAC β1-4 branch on the core structure of complex N-Glycans, which is the common substrate for other N -acetylglucosaminyltransferases, such as GnT-III and GnT-V. Our recent study indicates that the expression of GnT-IVa in Hca-F cells was much higher than that in Hepa1-6 cells, these two mouse hepatocarcinoma cell lines have high and no metastatic potential in lymph nodes respectively. To investigate the effects of GnT-IVa on the metastasis of hepatocarcinoma, exogenous GnT-IVa was introduced into Hepa1-6 cells, and on the other hand, the expression of GnT-IVa was down-regulated in Hca-F cells. The engineered overexpression of GnT-IVa in Hepa1-6 cells increased the antennary branches of complex N-glycans and reduced bisecting branches in vitro and in vivo , which leads to the increase in migration and metastatic capability of hepatocarcinoma cells. Conversely, down-regulated expression of GnT-IVa in Hca-F cells showed reduced tetra-antennary branches of N-Glycans, and significantly decreased the migration and metastatic capability. Furthermore, we found that the regulated GnT-IVa converts the heterogeneous N-glycosylated forms of CD147 in Hepa1-6 and Hca-F cells, and significantly changed the antennary oligosaccharide structures on CD147. These results suggest that GnT-IVa could be acting as a key role in migration and metastasis of mouse hepatocarcinoma cells through altering the glycosylation of CD147. These findings should be valuable in delineating the important function of GnT-IVa during the process of hepatocarcinoma growth and metastasis. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9414-1 Authors Jianhui Fan, Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, Dalian, 116044 China Shujing Wang, Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, Dalian, 116044 China Shengjin Yu, Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, Dalian, 116044 China Jingna He, Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, Dalian, 116044 China Weilong Zheng, Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, Dalian, 116044 China Jianing Zhang, Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, Dalian, 116044 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    We evaluated the carbohydrate preferences of the C-type lectin receptors (CLRs) SIGNR1, SIGNR3, and Langerin as pathogen-uptake receptors based on uptake of liposomes consisting of cholesterol, DPPC, and various neoglycolipids at molar ratios of 10:10:1 and 10:7:4, respectively, using non-phagocytic CHO cells that express these receptors transiently. SIGNR1-expressing cells ingested liposomes coated with neoglycolipids with terminal mannose residues, such as Man2-, Man3-, and Man5-DPPE, and with a terminal N -acetylglucosamine. SIGNR1 mediated uptake of Man3-DPPE-coated liposomes most efficiently. Uptake of liposomes with lower neoglycolipid content by SIGNR3- or Langerin-expressing cells was slight or negligible, but uptake into these cells was detected for liposomes with higher neoglycolipid content. SIGNR1-expressing cells clearly ingested liposomes coated with Lewis X antigen, whereas SIGNR3- or Langerin-expressing cells barely ingested these liposomes, even at the higher neoglycolipid content. In contrast, SIGNR3 or Langerin, but not SIGNR1, mediated uptake of liposomes coated with blood group H antigen. These results indicate that CLRs with similar carbohydrate-recognition characteristics have distinct properties as pathogen-uptake receptors for carbohydrate-decorated particles. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9406-1 Authors Yoko Kawauchi, Department of Applied Biochemistry, Tokai University, 4-1-1 Kita-kaname, Hiratsuka-shi, Kanagawa 259-1292, Japan Yasuhiro Kuroda, Department of Applied Biochemistry, Tokai University, 4-1-1 Kita-kaname, Hiratsuka-shi, Kanagawa 259-1292, Japan Naoya Kojima, Department of Applied Biochemistry, Tokai University, 4-1-1 Kita-kaname, Hiratsuka-shi, Kanagawa 259-1292, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Kashin-Beck Disease (KBD) is an endemic, chronic and degenerative osteoarthropathy principally occurring in children. The characteristic pathological change of KBD is chondrocyte necrosis in hyaline articular cartilage. Proteoglycans are one of the major components in the extracellular matrix of articular cartilage, and disrupted proteoglycan metabolism and loss of proteoglycans in articular cartilage from KBD patients has been observed. In this mini-review, we discuss the close relationship between chondrocyte death including necrosis and loss of proteoglycan, and its potential mechanism during KBD onset and development, which may provide new clues for KBD research. Content Type Journal Article Pages 1-8 DOI 10.1007/s10719-012-9421-2 Authors Siyuan Li, Key Laboratory of Environment and Genes Related to Diseases (Xi’an Jiaotong University), Ministry of Education, Xi’an, China 710061 Junling Cao, Key Laboratory of Environment and Genes Related to Diseases (Xi’an Jiaotong University), Ministry of Education, Xi’an, China 710061 Bruce Caterson, Connective Tissue Biology Laboratories, Division of Pathophysiology and Repair, School of Biosciences, Cardiff University, Cardiff, UK CF10 3AX Clare E. Hughes, Connective Tissue Biology Laboratories, Division of Pathophysiology and Repair, School of Biosciences, Cardiff University, Cardiff, UK CF10 3AX Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description: Erratum to: Carbohydrate to carbohydrate interaction in development process and cancer progression Content Type Journal Article Category Erratum Pages 1-1 DOI 10.1007/s10719-012-9422-1 Authors Kazuko Handa, Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122, USA Sen-itiroh Hakomori, Pacific Northwest Research Institute, 720 Broadway, Seattle, WA 98122, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The first step in the process of infections by the hepatitis C virus (HCV) is attachment to the host cell, which is assumed to be mediated by interaction of the envelope glycoproteins E1 and E2 with cell surface glycosaminoglycans. In this study, a variety of glycosaminoglycans, heparan sulfate (HS) from various bovine tissues as well as chondroitin sulfate (CS)/dermatan sulfate from bovine liver, were used to examine the direct interaction with recombinant E1 and E2 proteins. Intriguingly, among HS preparations from various bovine tissues, only liver HS strongly bound to both E1 and E2. Since HS from liver, which is the target tissue of HCV, contains highly sulfated structures compared to HS from other tissues, the present results suggest that HS-proteoglycan on the liver cell surface appears to be one of the molecules that define the liver-specific tissue tropism of HCV infection. The interaction assay with chemically modified heparin derivatives provided evidence that the binding of the viral proteins to heparin/HS is not only mediated by simple ionic interactions, but that the 6- O -sulfation and N -sulfation are important. Heparin oligosaccharides equal to or larger than 10-mer were required to inhibit the binding. Notably, a highly sulfated CS-E preparation from squid cartilage also strongly interacted with both viral proteins and inhibited the entry of pseudotype HCV into the target cells, suggesting that the highly sulfated CS-E might be useful as an anti-HCV drug. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9388-z Authors Fumi Kobayashi, Laboratory of Proteoglycan Signaling and Therapeutics, Hokkaido University Graduate School of Life Science, West-11, North-21, Kita-ku, Sapporo, 001-0021 Japan Shuhei Yamada, Laboratory of Proteoglycan Signaling and Therapeutics, Hokkaido University Graduate School of Life Science, West-11, North-21, Kita-ku, Sapporo, 001-0021 Japan Shuhei Taguwa, Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Yamada-oka 3-1, Suita-shi, Osaka 565-0871, Japan Chikako Kataoka, Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Yamada-oka 3-1, Suita-shi, Osaka 565-0871, Japan Satomi Naito, Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan Yoshiki Hama, Laboratory of Proteoglycan Signaling and Therapeutics, Hokkaido University Graduate School of Life Science, West-11, North-21, Kita-ku, Sapporo, 001-0021 Japan Hideki Tani, Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Yamada-oka 3-1, Suita-shi, Osaka 565-0871, Japan Yoshiharu Matsuura, Department of Molecular Virology, Research Institute for Microbial Diseases, Osaka University, Yamada-oka 3-1, Suita-shi, Osaka 565-0871, Japan Kazuyuki Sugahara, Laboratory of Proteoglycan Signaling and Therapeutics, Hokkaido University Graduate School of Life Science, West-11, North-21, Kita-ku, Sapporo, 001-0021 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    In an effort to prime our mass spectrometry (MS)-based sulfoglycomic mapping platform technology for facile identification of sulfated lacdiNAc (GalNAcβ1-4GlcNAcβ1-), we have re-examined the N-glycans of bovine thyroid stimulating hormone. We showed that MALDI-MS mapping of permethylated glycans in negative ion mode can give an accurate representation of the sulfated glycans and, through MS/MS, diagnostic ions can be derived that we can collectively define the presence of a terminal sulfated lacdiNAc moiety at high sensitivity. Based on these ions, which can also be produced by nanoESI-MS n , we demonstrated that the glycome of an ovarian carcinoma cell line, RMG-1, comprises a high abundance of sulfated lacdiNAc epitopes carried on multiantennary complex type N-glycans alongside fucosylated, sialylated and/or sulfated lacNAc antennae. This represents the first report of a natural glycomic occurrence of sulfated lacdiNAc on a cell line, as opposed to other better-characterized presence on secreted glycoproteins from a handful of sources. It is anticipated that with improved methods of detection such as that developed in this work, we are likely to identify a wider occurrence of sulfated lacdiNAc and be able to more accurately delineate the regulatory mechanism dictating the choice of a cell type in synthesizing sulfated, sialylated, fucosylated and/or non-substituted lacdiNAc. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9396-z Authors Shin-Yi Yu, Institute of Biological Chemistry, Academia Sinica, 128, Academia Road Sec 2, Nankang, Taipei, 115 Taiwan Lan-Yi Chang, Institute of Biological Chemistry, Academia Sinica, 128, Academia Road Sec 2, Nankang, Taipei, 115 Taiwan Chu-Wen Cheng, Institute of Biochemical Sciences, National Taiwan University, Taipei, 106 Taiwan Chi-Chi Chou, Core Facilities for Protein Structural Analysis, NCFPB, Academia Sinica, Taipei, 115 Taiwan Michiko N. Fukuda, Tumor Microenvironmental Program, Cancer Center, Sanford-Burnham Medical Research Institute, La Jolla, CA 920137, USA Kay-Hooi Khoo, Institute of Biological Chemistry, Academia Sinica, 128, Academia Road Sec 2, Nankang, Taipei, 115 Taiwan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Heparan sulfate proteoglycan (HSPG), such as glypican, plays a role as a co-receptor for growth factor to influence cells proliferation. However the mechanism is still vague. Micro-RNAs (miRNAs) regulate cell proliferation. Their capacity to direct the translation and stability of targeted transcripts can dramatically influence cellular physiological function. To explore how the function of glypican is regulated involved in cell proliferation, glypican-4 was chosen with a bioinformatics search identifying targeting seed sequences for miR-125a within the 3′-untranslated regions (3′UTR). Indeed, luciferase constructs containing the 3′UTR of glypican-4 demonstrated around 54 % less activity in miR-125a ex pressing cells relative to the controls. The expression of glypican-4 at both the transcript and protein level was down-regulated by transition trasfection of miR-125a in the human embryonic kidney cell line 293T (HEK293T). Although cell proliferation of HEK293T was not influenced by the silence of glypican-4, DNA synthesis in response to FGF2 in the cells was attenuated by knockdown of glypican-4 using siRNA technique. Further study showed that phosphorylation of ERK 1/2 and AKT was suppressed by overexpressing miR-125a , whereas the suppressed MAPK and AKT signaling could be recovered by anti- miR-125a treatment. Both DNA synthesis and cell proliferation were impaired by the inhibitor of ERK 1/2 signaling. MTT assay demonstrated that the cell proliferation was impaired by miR-125a overexpression, however, rescued by anti -miR-125a in HEK293T cells. These results disclosed new function of miR-125a by targeting gene glypican-4 in cell growth process and illustrated the feasibility of using miRNAs as a therapeutic strategy to suppress cells proliferation. Content Type Journal Article Pages 1-9 DOI 10.1007/s10719-012-9387-0 Authors Chao Feng, Glycochemistry & Glycobiology Lab, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203 China Jie Li, Glycochemistry & Glycobiology Lab, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203 China Jinlan Ruan, Hubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, College of Pharmacy, Tongji Medical Center of Huazhong University of Science and Technology, Wuhan, 430030 China Kan Ding, Glycochemistry & Glycobiology Lab, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Aberrant glycosylation is a characteristic feature of cancer cells. In particular, altered sialylation is closely associated with malignant properties, including invasiveness and metastatic potential. To elucidate the molecular mechanisms underlying the aberrancy, our studies have focused on mammalian sialidase, which catalyzes the removal of sialic acid residues from glycoproteins and glycolipids. The four types of mammalian sialidase identified to date show altered expression and behave in different manners during carcinogenesis. The present review briefly summarizes results on altered expression of sialidases and their possible roles in cancer progression. These enzymes are indeed factors defining cancer malignancy and thus potential targets for cancer diagnosis and therapy. Content Type Journal Article Pages 1-11 DOI 10.1007/s10719-012-9394-1 Authors Taeko Miyagi, Division of Cancer Glycosylation Research, Institute of Molecular Biomembrane and Glycobiology, Tohoku Pharmaceutical University, 981-8558 Sendai, Japan Kohta Takahashi, Division of Cancer Glycosylation Research, Institute of Molecular Biomembrane and Glycobiology, Tohoku Pharmaceutical University, 981-8558 Sendai, Japan Keiko Hata, Division of Cancer Glycosylation Research, Institute of Molecular Biomembrane and Glycobiology, Tohoku Pharmaceutical University, 981-8558 Sendai, Japan Kazuhiro Shiozaki, Laboratory of Marine Biochemistry, Faculty of Fisheries, Kagoshima University, Kagoshima, 890-0056 Japan Kazunori Yamaguchi, Division of Molecular and Cellular Oncology, Miyagi Cancer Center Research Institute, Natori, 981-1293 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The majority of all proteins are glycosylated and glycans have numerous important structural, functional and regulatory roles in various physiological processes. While structure of the polypeptide part of a glycoprotein is defined by the sequence of nucleotides in the corresponding gene, structure of a glycan part results from dynamic interactions between hundreds of genes, their protein products and environmental factors. The composition of the glycome attached to an individual protein, or to a complex mixture of proteins, like human plasma, is stable within an individual, but very variable between individuals. This variability stems from numerous common genetic polymorphisms reflecting in changes in the complex biosynthetic pathway of glycans, but also from the interaction with the environment. Environment can affect glycan biosynthesis at the level of substrate availability, regulation of enzyme activity and/or hormonal signals, but also through gene-environment interactions. Epigenetics provides a molecular basis how the environment can modify phenotype of an individual. The epigenetic information (DNA methylation pattern and histone code) is especially vulnerable to environmental effects in the early intrauterine and neo-natal development and many common late-onset diseases take root already at that time. The evidences showing the link between epigenetics and glycosylation are accumulating. Recent progress in high-throughput glycomics, genomics and epigenomics enabled first epidemiological and genome-wide association studies of the glycome, which are presented in this mini-review. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9397-y Authors Vlatka Zoldoš, University of Zagreb, Faculty of Science, Horvatovac 102a, Zagreb, Croatia Mislav Novokmet, Genos Ltd, Glycobiology Laboratory, Planinska 1, Zagreb, Croatia Ivona Bečeheli, Genos Ltd, Glycobiology Laboratory, Planinska 1, Zagreb, Croatia Gordan Lauc, Genos Ltd, Glycobiology Laboratory, Planinska 1, Zagreb, Croatia Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    M. tuberculosis GlmU is a bifunctional enzyme with acetyltransferase activity in C-terminus and uridyltransferase activity in N-terminus, and it is involved in the biosynthesis of glycosyl donor UDP- N -acetylglucosamine (UDP-GlcNAc). The crystal structure of M. tuberculosis GlmU clearly determines the active site and catalytic mechanism of GlmU uridyltransferase domain but not succeed in GlmU acetyltransferase domain. Sequence comparison analysis revealed highly conserved amino acid residues in the C-terminus between M. tuberculosis GlmU and GlmU enzymes from other bacteria. To find the essential amino acids related to M. tuberculosis GlmU acetyltransferase activity, we substituted 10 conserved amino acids in the acetyltransferase domain of M. tuberculosis GlmU by site-directed mutagenesis. All the mutant GlmU proteins were largely expressed in soluble and purified by affinity chromatography. Enzyme assays showed that K362A, H374A, Y398A and W460A mutants abolished more than 90 % activity of M. tuberculosis GlmU acetyltransferase and totally lost the affinity with two substrates, suggesting the potential substrate-binding functions. However, K403A, S416A, N456A and E458A mutants exhibited decreased GlmU acetyltransferase activity and lower kinetic parameters, probably responsible for substrate releasing by conformation shifting. Content Type Journal Article Pages 1-7 DOI 10.1007/s10719-012-9402-5 Authors Yan Zhou, Department of Biochemistry and Molecular Biology, Dalian Medical Universtiy, Dalian, 116044 China Wendan Yu, Department of Biochemistry and Molecular Biology, Dalian Medical Universtiy, Dalian, 116044 China Qi Zheng, Department of Biochemistry and Molecular Biology, Dalian Medical Universtiy, Dalian, 116044 China Yi Xin, Department of Biotechnology, Dalian Medical University, Dalian, 116044 China Yufang Ma, Department of Biochemistry and Molecular Biology, Dalian Medical Universtiy, Dalian, 116044 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Tetrasaccharide cap present in lipophosphoglycan of the Leishmania donovani responsible for visceral Leishmaniaisis is synthesized as a fully protected propargyl glycoside. AuBr 3 mediated selective glycosylation of propargyl 1,2-orthoester in the presence of propargyl glycoside is employed as a key step to obtain propargyl containing oligomers. Further, propargyl tetrasaccharide is connected with a long chain hydrocarbon containing azidothiol functionality situated at two terminal ends via ‘click’ reaction. Content Type Journal Article Pages 1-10 DOI 10.1007/s10719-012-9400-7 Authors Gopalsamy Sureshkumar, Department of Chemistry, Indian Insititute of Science Education & Research, Pune, 411 008 India Srinivas Hotha, Department of Chemistry, Indian Insititute of Science Education & Research, Pune, 411 008 India Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    With the booming development of glycobiology and glycochemistry, more and more structures of tumor-associated carbohydrate antigens (TACAs) are identified. Their broad expression and high specificity in cancer make them important targets to develop cancer vaccines or immunotherapies. However, most of the TACAs are T cell-independent antigens, they cannot elicit a powerful enough immune response to prevent or treat cancer. Immunotolerance and immunosuppression are more easily induced due to their endogenous properties and the declining immunity of the patients. This review summarizes the recent efforts to overcome these obstacles: coupling the carbohydrate antigens to proper carriers such as proteins or some small molecule carriers, and chemically modifying the structures of the TACAs to enhance the immunogenicity of TACAs and break the immunotolerance. Content Type Journal Article Pages 1-13 DOI 10.1007/s10719-012-9399-9 Authors Chang-Cheng Liu, State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Xue Yuan Road No. 38, Beijing, 100191 China Xin-Shan Ye, State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Xue Yuan Road No. 38, Beijing, 100191 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description: International Glycoconjugate Organization (IGO) award for 2013 Content Type Journal Article Pages 1-2 DOI 10.1007/s10719-012-9409-y Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Nucleotide sugar transporters play critical roles in glycosylation of proteins, lipids and proteoglycans, which are essential for organogenesis, development, mammalian cellular immunity and pathogenicity of human pathogenic agents. Functional deficiencies of these transporters result in global defects of glycoconjugates, which in turn lead to a diversity of biochemical, physiological and pathological phenotypes. In this short review, we will highlight human and bovine diseases caused by mutations of these transporters. Content Type Journal Article Pages 1-6 DOI 10.1007/s10719-012-9375-4 Authors Li Liu, Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Evans-E438, 72 East Concord Street, Boston, MA 02118, USA Carlos B. Hirschberg, Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Evans-E438, 72 East Concord Street, Boston, MA 02118, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    The free-living nematode Caenorhabditis elegans is a well-characterized eukaryotic model organism. Recent glycomic analyses of the glycosylation potential of this worm revealed an extremely high structural variability of its N-glycans. Moreover, the glycan patterns of each developmental stage appeared to be unique. In this study we have determined the N-glycan profiles of wild-type embryos in comparison to mutant embryos arresting embryogenesis early before differentiation and causing extensive transformations of cell identities, which allows to follow the diversification of N-glycans during development using mass spectrometry. As a striking feature, wild-type embryos obtained from liquid culture expressed a less heterogeneous oligosaccharide pattern than embryos recovered from agar plates. N-glycan profiles of mutant embryos displayed, in part, distinct differences in comparison to wild-type embryos suggesting alterations in oligosaccharide trimming and processing, which may be linked to specific cell fate alterations in the embryos. Content Type Journal Article Pages 135-145 DOI 10.1007/s10719-012-9371-8 Authors Hildegard Geyer, Institute of Biochemistry, Faculty of Medicine, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany Martin Schmidt, Institute of Biochemistry, Faculty of Medicine, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany Matthias Müller, Institut für Genetik, Technische Universität Braunschweig Carolo Wilhelmina, Spielmannstrasse 7, 38106 Braunschweig, Germany Ralf Schnabel, Institut für Genetik, Technische Universität Braunschweig Carolo Wilhelmina, Spielmannstrasse 7, 38106 Braunschweig, Germany Rudolf Geyer, Institute of Biochemistry, Faculty of Medicine, University of Giessen, Friedrichstrasse 24, 35392 Giessen, Germany Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Numbers 2-3

Description:    Using printed glycan array (PGA) we compared the results of antibody profiling in undiluted, moderately (1:15) and highly (1:100) diluted human blood serum. Undiluted serum is suitable for studying blood as a tissue in its native state, whereas to study the serum of newborns or small animals one usually has to dilute the starting material in order to have sufficient volume for PGA experimentation. The PGA used in this study allows for the use of whole serum without modifications to the protocol, and the background is surprisingly low. Antibodies profiles observed in undiluted serum versus 1:15 dilution were similar, with only a limited number of new signals identified in the undiluted serum. However, unexpected irregularities were found when IgG and IgM are measured separately, namely, at a 1:15 dilution more intensive IgG signals for many glycans are observed. We believe that in conditions of moderate dilution IgG and IgM antibodies can compete with each other for antigen and as a result, the higher affinity anti-glycan IgGs give rise to more intense signals. Therefore depending on the purpose, different dilutions of serum will be optimal: in competitive 1:15 conditions the observed IgG/IgM ratio corresponds to their titer, whereas at 1:100 dilution the measured ratio corresponds to real molar concentration of IgG and IgM. Content Type Journal Article Category GlycoArray Section Pages 87-91 DOI 10.1007/s10719-011-9368-8 Authors Nadezhda Shilova, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Maxim Navakouski, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Nailya Khasbiullina, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Ola Blixt, Copenhagen Center for Glycomics, Institute of Cellular and Molecular Medicine, Panum Institute, University of Copenhagen, Blegdamsvej 3, DK-2200 Copenhagen, Denmark Nicolai Bovin, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Numbers 2-3

Description:    In the milk of marsupials, oligosaccharides usually predominate over lactose during early to mid lactation. Studies have shown that tammar wallaby milk contains a major series of neutral galactosyllactose oligosaccharides ranging in size from tri- to at least octasaccharides, as well as β(1-6) linked N -acetylglucosamine-containing oligosaccharides as a minor series. In this study, acidic oligosaccharides were purified from red kangaroo milk and characterized by 1 H-nuclear magnetic resonance spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, to be as follows: Neu5Ac(α2-3)Gal(β1-4)Glc (3′-SL), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-4)Glc (sialyl 3′-galactosyllactose), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Neu5Ac(α2-3)Gal(β1-3)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Neu5Ac(α2-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose a), Gal(β1-3)[Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose b), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc, Gal(β1-3)(-3- O -sulfate)Gal(β1-3)Gal(β1-4)Glc, Gal(β1-3)(-3- O -sulfate)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Gal(β1-3)(-3- O -sulfate)Gal(β1-3)Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc, Gal(β1-3)(-3- O -sulfate)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc, Gal(β1-3)(-3- O -sulfate)Gal(β1-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc. These acidic oligosaccharides were shown to be sialylated or sulfated in the non-reducing ends to the major linear and the minor branched series of neutral oligosaccharides of tammar wallaby milk. Content Type Journal Article Pages 147-156 DOI 10.1007/s10719-012-9372-7 Authors Tatsuro Anraku, Graduate School of Food Hygiene, Obihiro University of Agriculture & Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan Kenji Fukuda, Graduate School of Food Hygiene, Obihiro University of Agriculture & Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan Tadao Saito, Graduate School of Agriculture, Tohoku University, Sendai, Miyagi 981-8555, Japan Michael Messer, School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, NSW 2006, Australia Tadasu Urashima, Graduate School of Food Hygiene, Obihiro University of Agriculture & Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Numbers 2-3

Description:    Surfactant protein A (SP-A), which is a lung innate immune system component, is known to bind glycolipids present at the cell surface of a mycobacterial pathogen. Lipoarabinomannan (LAM), a component of mycobacterial thick, waxy cell wall, is one of the glycolipid ligands for SP-A. In order to assess binding of synthetic glycolipids with SP-A and the glycosidic linkage preferences for the interaction, β-arabinofuranoside trisaccharide glycolipids constituted with β-(1→2), β-(1→3) and β-(1→2), β-(1→5) linkages relevant to LAM were synthesized through chemical glycosylations. The efficacies of synthetic glycolipids to interact with SP-A were assessed by using the surface plasmon resonance (SPR) technique, from which association-dissociation rate constants and equilibrium binding constants were derived. The equilibrium binding constants of the interaction of two constitutionally varying β-arabinofuranoside glycolipids with SP-A were found to be in the millimolar range. A comparison of the results with few α-anomeric arabinofuranoside glycolipids showed that glycolipids with β-anomeric linkages were having relatively lower equilibrium binding constants than those with α-anomeric linkages in binding to the protein, whereas oligosaccharides alone, without lipidic chains, exhibited higher equilibrium binding constants. Further, the synthetic compounds inhibited the growth of mycobacteria and affected sliding motilities of the bacteria, although to an extent relatively lesser than that of synthetic compounds constituted with α-anomeric linkages. Content Type Journal Article Pages 107-118 DOI 10.1007/s10719-012-9369-2 Authors Kottari Naresh, Department of Organic Chemistry, Indian Institute of Science, Bangalore, 560 012 India Prakash Gouda Avaji, Department of Organic Chemistry, Indian Institute of Science, Bangalore, 560 012 India Krishnagopal Maiti, Department of Organic Chemistry, Indian Institute of Science, Bangalore, 560 012 India Binod K. Bharati, Molecular Biophysics Unit, Indian Institute of Science, Bangalore, 560 012 India Kirtimaan Syal, Molecular Biophysics Unit, Indian Institute of Science, Bangalore, 560 012 India Dipankar Chatterji, Molecular Biophysics Unit, Indian Institute of Science, Bangalore, 560 012 India Narayanaswamy Jayaraman, Department of Organic Chemistry, Indian Institute of Science, Bangalore, 560 012 India Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Numbers 2-3

Description:    A carbohydrate-binding module from family 13 (CBM13), appended to the catalytic domain of endo-1,3-β-glucanase from Cellulosimicrobium cellulans , was overexpressed in E. coli , and its interactions with β-glucans, laminarin and laminarioligosaccharides, were analyzed using surface plasmon resonance biosensor and isothermal titration calorimetry. The association constants for laminarin and laminarioligosaccharides were determined to be approximately 10 6  M −1 and 10 4  M −1 , respectively, indicating that 2 or 3 binding sites in the α-, β-, and γ-repeats of CBM13 are involved in laminarin binding in a cooperative manner. The binding avidity is approximately 2-orders higher than the monovalent binding affinity. Mutational analysis of the conserved Asp residues in the respective repeats showed that the α-repeat primarily contributes to β-glucan binding. A Trp residue is predicted to be exposed to the solvent only in the α-repeat and would contribute to β-glucan binding. The α-repeat bound β-glucan with an affinity of approximately 10 4  M −1 , and the other repeats additionally bound laminarin, resulting in the increased binding avidity. This binding is unique compared to the recognition mode of another CBM13 from Streptomyces lividans xylanase. Content Type Journal Article Pages 77-85 DOI 10.1007/s10719-011-9366-x Authors Tomonari Tamashiro, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, 1-5 Hangi-cho, Shimogamo, Sakyo-ku, Kyoto, 606-8522 Japan Yoichi Tanabe, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, 1-5 Hangi-cho, Shimogamo, Sakyo-ku, Kyoto, 606-8522 Japan Teikichi Ikura, Graduate School of Biomedical Science, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510 Japan Nobutoshi Ito, Graduate School of Biomedical Science, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510 Japan Masayuki Oda, Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, 1-5 Hangi-cho, Shimogamo, Sakyo-ku, Kyoto, 606-8522 Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Number 1

Description:    Endoplasmic reticulum α-1,2 mannosidase I (ERManI) is an enzyme, which removes α(1-2) linked mannoses from asparagine-linked oligosaccharides on glycoproteins in the endoplasmic reticulum (ER). ERManI preferentially removes one α(1-2) linked mannose from B-chain of Man 9 GlcNAc 2 . When glycoproteins fail to achieve properly folding, increased removal of α(1-2) linked mannoses on their oligosaccharides is induced and leads them to be disposed and degraded by ER-associated degradation pathway. However, it is still inconclusive whether accelerated removal of α(1-2) linked mannoses on those glycoproteins is catalyzed by the α-1,2 mannosidase I, proteins similar to mannosidase I [ e.g. ER degradation-enhancing α-1,2 mannosidase-like protein (EDEM)], or both of them. Therefore, to approach this issue, we have investigated its in vitro activities using various oligosaccharides and glycoproteins as substrates. A recombinant form of human ERManI (hERManI) was prepared by using Escherichia coli . First, the enzyme generated Man 6 GlcNAc 2 -PA and Man 5 GlcNAc 2 -PA from 100 μM Man 9 GlcNAc 2 -PA after a one-hour reaction. Second, we have exposed bovine thyroglobulin and soybean agglutinin to denaturing conditions, e.g. 8 M urea, and used those glycoproteins as substrates. Sugar moieties were released from the reactant by PNGase F and their structures and amounts were elucidated by HPLC analysis. Intriguingly, the enzyme was shown to remove mannoses from bovine thyroglobulin and soybean agglutinin to larger extents when they were exposed to a denaturant. Therefore, our results suggested that hERManI could recognize tertiary and/or quaternary structures of glycoproteins and remove more α-1,2 linked mannoses from misfolded glycoproteins in living cells. Content Type Journal Article Pages 35-45 DOI 10.1007/s10719-011-9362-1 Authors Jun-ichi Aikawa, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Ichiro Matsuo, Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan Yukishige Ito, RIKEN Advanced Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Number 1

Description:    Phosphorylated modification of a polysaccharide obtained from Radix Hedysari (RHP) was studied. Three phosphorylated polysaccharides (RHPP) with variable degrees of substitution (DS p ) were obtained with 4-dimethylaminopyridine (DMAP) and N , N ′ Dicyclocarbodiimide (DCC) as catalyst. The structures of RHPP were characterized by FT-IR spectra and 13 C NMR spectra. Depending on different reaction time, RHPP showed different DS p ranging from 0.30 to 0.66, and different Mw ranging from 86.6 to 89.7 KDa. Compared with RHP, RHPP exhibited superior antioxidant activities in vitro , which indicated that phosphorylated modification could enhance antioxidant activities of RHP. Furthermore, it was obvious that the DS p had a significant effect on the antioxidant activity. Content Type Journal Article Pages 1-6 DOI 10.1007/s10719-012-9377-2 Authors Dongfeng Wei, School of Basic Medical Science, Lanzhou University, Lanzhou, 730000 China Weidong Cheng, School of Basic Medical Science, Lanzhou University, Lanzhou, 730000 China Yanxia Wei, School of Basic Medical Science, Lanzhou University, Lanzhou, 730000 China Lifeng Zhang, School of Basic Medical Science, Lanzhou University, Lanzhou, 730000 China Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Duffy antigen/receptor for chemokines (DARC) is a glycosylated seven-transmembrane protein acting as a blood group antigen, a chemokine binding protein and a receptor for Plasmodium vivax malaria parasite. It is present on erythrocytes and endothelial cells of postcapillary venules. The N-terminal extracellular domain of the Duffy glycoprotein carries Fy a /Fy b blood group antigens and Fy6 linear epitope recognized by monoclonal antibodies. Previously, we have shown that recombinant Duffy protein expressed in K562 cells has three N- linked oligosaccharide chains, which are mainly of complex-type. Here we report a one-step purification method of Duffy protein from human erythrocytes. DARC was extracted from erythrocyte membranes in the presence of 1% n -dodecyl-β-D-maltoside (DDM) and 0.05% cholesteryl hemisuccinate (CHS) and purified by affinity chromatography using immobilized anti-Fy6 2C3 mouse monoclonal antibody. Duffy glycoprotein was eluted from the column with synthetic DFEDVWN peptide containing epitope for 2C3 monoclonal antibody. In this single-step immunoaffinity purification method we obtained highly purified DARC, which migrates in SDS-polyacrylamide gel as a major diffuse band corresponding to a molecular mass of 40–47 kDa. In ELISA purified Duffy glycoprotein binds anti-Duffy antibodies recognizing epitopes located on distinct regions of the molecule. Results of circular dichroism measurement indicate that purified DARC has a high content of α-helical secondary structure typical for chemokine receptors. Analysis of DARC glycans performed by means of lectin blotting and glycosidase digestion suggests that native Duffy N -glycans are mostly triantennary complex-type, terminated with α2-3- and α2-6-linked sialic acid residues with bisecting GlcNAc and α1-6-linked fucose at the core. Content Type Journal Article Pages 93-105 DOI 10.1007/s10719-011-9367-9 Authors Magdalena Grodecka, Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, 53-114 Wrocław, Poland Olivier Bertrand, Institut National de la Santé et de la Recherche Médicale, UMR_S 665, F-75015 Paris, France Ewa Karolak, Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, 53-114 Wrocław, Poland Marek Lisowski, Faculty of Chemistry, University of Wrocław, F. Joliot-Curie 14, 50-383 Wrocław, Poland Kazimiera Waśniowska, Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, R. Weigla 12, 53-114 Wrocław, Poland Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Numbers 2-3

Description:    Bacillus anthracis toxins may be attenuated if macrophages could neutralize toxins upon contact or exposure. Glycoconjugate-bearing polymers, which have been shown to bind to Bacillus spores, were tested for recognition and binding of protective antigen (PA), lethal factor (LF), and edema factor (EF) toxins. We have demonstrated modulation of macrophage activity following exposure to these toxins. Without glycoconjugate (GC) activation, murine macrophages were killed by Bacillus toxins. GCs were shown to have a protective influence, sparing macrophages from toxin-induced cell death, as shown by increased macrophage cell viability based on trypan blue assay. Increased levels of inducible nitric oxide (NO) production by macrophages in presence of GCs suggest that GCs provide an activation signal for macrophages and stimulate their function. Results hint to GCs that promote neutralization of Bacillus toxins, block toxin-induced macrophage death, while increasing macrophage activation. Polymeric GCs may suggest novel approaches to improve existing or develop new vaccines as well as immunotherapeutics. Content Type Journal Article Pages 25-33 DOI 10.1007/s10719-011-9360-3 Authors Olga Tarasenko, Department of Biology, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Ashley Scott, Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR, USA Lee Soderberg, Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, AR, USA Pierre Alusta, Bioinformatics Program, University of Arkansas at Little Rock, 2801 South University Ave., Little Rock, AR 72204, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Number 1

Description: Obituary: Nathan Sharon (1925–2011) Content Type Journal Article Pages 499-500 DOI 10.1007/s10719-011-9361-2 Authors Harry Schachter, Molecular Structure and Function Program, Hospital for Sick Children, 555 University Avenue, Toronto, Ont M5G 1X8, Canada Hans Vliegenthart, Bijvoet Center, Utrecht University, Padualaan 8, NL 3584 CH Utrecht, The Netherlands Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 8-9

Description:    Glycosphingolipids (GSLs) are well known ubiquitous constituents of all eukaryotic cell membranes, yet their normal biological functions are not fully understood. As with other glycoconjugates and saccharides, solid phase display on microarrays potentially provides an effective platform for in vitro study of their functional interactions. However, with few exceptions, the most widely used microarray platforms display only the glycan moiety of GSLs, which not only ignores potential modulating effects of the lipid aglycone, but inherently limits the scope of application, excluding, for example, the major classes of plant and fungal GSLs. In this work, a prototype “universal” GSL-based covalent microarray has been designed, and preliminary evaluation of its potential utility in assaying protein-GSL binding interactions investigated. An essential step in development involved the enzymatic release of the fatty acyl moiety of the ceramide aglycone of selected mammalian GSLs with sphingolipid N -deacylase (SCDase). Derivatization of the free amino group of a typical lyso-GSL, lyso-G M1 , with a prototype linker assembled from succinimidyl-[( N -maleimidopropionamido)-diethyleneglycol] ester and 2-mercaptoethylamine, was also tested. Underivatized or linker-derivatized lyso-GSL were then immobilized on N -hydroxysuccinimide- or epoxide-activated glass microarray slides and probed with carbohydrate binding proteins of known or partially known specificities ( i.e. , cholera toxin B-chain; peanut agglutinin, a monoclonal antibody to sulfatide, Sulph 1; and a polyclonal antiserum reactive to asialo-G M2 ). Preliminary evaluation of the method indicated successful immobilization of the GSLs, and selective binding of test probes. The potential utility of this methodology for designing covalent microarrays that incorporate GSLs for serodiagnosis is discussed. Content Type Journal Article Category Glycoarray Section Pages 1-12 DOI 10.1007/s10719-011-9359-9 Authors Emma Arigi, Department of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3, 2200 Copenhagen N, Denmark Ola Blixt, Department of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3, 2200 Copenhagen N, Denmark Karsten Buschard, Rigshospitalet, Bartholin Institute, Blegdamsvej 9, 2100 Copenhagen OE, Denmark Henrik Clausen, Department of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3, 2200 Copenhagen N, Denmark Steven B. Levery, Department of Cellular and Molecular Medicine, University of Copenhagen, Blegdamsvej 3, 2200 Copenhagen N, Denmark Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Number 1

Description:    The structures of the branched capsular polysaccharides of group B streptococcus type III (GBSIIIPS) and Streptococcus pneumoniae type 14 (Pn14PS) are identical apart from the (α2→3)-linked sialic acid in the side chains of GBSIIIPS. The present study tries to determine the minimal epitope in GBSIIIPS, using both a panel of anti-Pn14PS mouse sera and sera of humans vaccinated with either Pn14PS or GBSIIIPS. Type-specific Pn14PS antibodies that recognize the branched structure of Pn14PS have a low affinity for the native GBSIIIPS. Desialylation of GBSIIIPS results in dramatically higher affinity of anti-Pn14PS antibodies. Epitope specific anti-Pn14PS mouse antibodies and human sera of PCV7 vaccinees only recognized structures with the branching element -Glc-(Gal-)GlcNAc-, in particular -Gal-Glc-(Gal-)GlcNAc- in Pn14PS. On the other hand anti-GBSIIIPS human antibodies recognize predominantly the linear structure in the backbone of Pn14PS or GBSIIIPS, i.e. , -Glc-GlcNAc-Gal-. This difference in antigenicity of Pn14PS and GBSIIIPS is in agreement with the difference in flexibility of the two polysaccharides caused by the presence or absence of sialic acid. Content Type Journal Article Pages 557-562 DOI 10.1007/s10719-011-9354-1 Authors Dodi Safari, Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands Huberta A. T. Dekker, Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands Ger T. Rijkers, Department of Medical Microbiology and Immunology, St. Antonius Hospital, Nieuwegein, The Netherlands Arie van der Ende, Department of Medical Microbiology and the Reference Laboratory for Bacterial Meningitis, Academic Medical Center, Amsterdam, The Netherlands Johannis P. Kamerling, Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Utrecht, The Netherlands Harm Snippe, Department of Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 8-9

Description:    The synthetic glycopeptides are interesting model systems to study the effect of O -glycosylation in modulating their function and structure. A series of glycosylated analogs of two antibacterial peptides, formaecin I and drosocin, were synthesized by varying the nature of sugar and its linkage with bioactive peptides to understand the influence of structure variation of glycosylation on their antibacterial activities. Higher antibacterial activities of all glycopeptides compared to their respective non-glycosylated counterparts emphasize in part the importance of sugar moieties in functional implications of these peptides. The consequences of the unique differences among the analogs were apparent on their antibacterial activities but not evident structurally by circular dichroism studies. We have shown that differently glycosylated peptides exhibit differential effect among each other when tested against several Gram-negative bacterial strains. The change of monosaccharide moiety and/or its anomeric configuration in formaecin I and drosocin resulted into decrease in the antibacterial activity in comparison to that of the native glycopeptide, but the extent of decrease in antibacterial activity of glycosylated drosocin analogs was less. Probably, the variation in peptide conformation arising due to topological dissimilarities among different sugars in the same peptide resulting in possible modulation in binding properties appears to be responsible for differences in their antibacterial activities. Indeed, these effects of glycosylation are found to be sequence-specific and depend in the milieu of amino acid residues. Interestingly, none of the carbohydrate variants affected the basic property of these peptides, which is non-hemolytic and non-toxicity to eukaryotic cells. Content Type Journal Article Pages 537-555 DOI 10.1007/s10719-011-9353-2 Authors Sariya Talat, Structural Biology Unit, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110 067 India Menithalakshmi Thiruvikraman, Structural Biology Unit, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110 067 India Saroj Kumari, Structural Biology Unit, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110 067 India Kanwal J. Kaur, Structural Biology Unit, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, 110 067 India Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 8-9

Description:    Using an example of Galβ1-3GlcNAc (Le C ) related glycans, we here demonstrate a risk of data misinterpretation when polyclonal antibodies are probed for their glycan-binding specificities with help of a printed glycan array (PGA). Affinity isolation of antibodies from human serum using Le C -Sepharose or 3′-O-SuLe C -Sepharose in conditions of excess of the adsorbents generated identical material regardless of the affinity ligand, with the antibodies equally capable of binding to Le C and to 3′-O-SuLe C disaccharides, as well as to 3′-O-SiaLe C trisaccharide. More detailed profiling has shown that the isolated antibodies bind to the inner part of Galβ1-3GlcNAc disaccharide. We therefore conclude that serum does not contain different subsets of antibodies specific either to Le C or to 3′-O-SuLe C , despite their visibly different binding signals to these glycans on PGA. Content Type Journal Article Category Glycoarray Section Pages 501-505 DOI 10.1007/s10719-011-9355-0 Authors Polina Obukhova, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Vladimir Piskarev, Glycoseparations, Moscow, Russia Vyacheslav Severov, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Galina Pazynina, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Alexander Tuzikov, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Maxim Navakouski, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Nadezhda Shilova, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Nicolai Bovin, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 8-9

Description:    A convergent synthesis of the tetrasaccharide repeating unit of the O -antigen of the verotoxin producing E. coli O176 has been achieved in excellent yield adopting a [2 + 2] block glycosylation strategy. The β-D-mannosidic moiety of the tetrasaccharide was prepared from β-D-glucoside and α-D-galactosamine moiety was derived from D-galactal. The tetrasaccharide was synthesized as its 2-trimethylsilylethyl glycoside in excellent yield. All intermediate steps are high yielding. Figure  A convergent synthetic strategy of the tetrasaccharide repeating unit corresponding to the O -antigen of verotoxin-producing Escherichia coli has been successfully developed using stereoselective [2 + 2] block glycosylation technique. Content Type Journal Article Pages 519-524 DOI 10.1007/s10719-011-9351-4 Authors Goutam Guchhait, Molecular Medicine Division, Bose Institute, P-1/12, C.I.T. Scheme VII M, Kolkata, 700054 India Anup Kumar Misra, Molecular Medicine Division, Bose Institute, P-1/12, C.I.T. Scheme VII M, Kolkata, 700054 India Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 8-9

Description: XXI International Symposium on Glycoconjugates Content Type Journal Article Pages 197-369 DOI 10.1007/s10719-011-9334-5 Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 5

Description:    Inner core lipopolysaccharide (LPS) has been shown to be conserved in the majority of veterinary strains from the species Mannheimia haemolytica , Actinobacillus pleuropneumoniae and Pasteurella multocida and as such is being considered as a possible vaccine antigen. The proof-in-principle that a LPS-based antigen could be considered as a vaccine candidate has been demonstrated from studies with monoclonal antibodies raised to the inner core LPS of Mannheimia haemolytica , which were shown to be both bactericidal and protective in a mouse model of disease. In this study we confirm and extend the candidacy of the inner core LPS by demonstrating that it is possible to elicit functional antibodies against Mannheimia haemolytica wild-type strains following immunisation of rabbits with glycoconjugates elaborating the conserved inner core LPS antigen. The present study describes a conjugation strategy that uses amidases produced by Dictyostelium discoideum , targeting the amino functionality created by the amidase activity as the attachment point on the LPS molecule. To protect the amino functionality on the phosphoethanolamine (PEtn) residue of the inner core, we developed a novel blocking and unblocking strategy with t-butyl oxycarbonyl. A maleimide-thiol linker strategy with the thiol linker on the carboxyl residues of the carrier protein and the maleimide linker on the carbohydrate resulted in a high loading of carbohydrates per carrier protein. Immunisation derived antisera from rabbits recognised fully extended Mannheimia haemolytica LPS and whole cells from serotypes 1 and 2, despite a somewhat immunodominant response to the linkers also being observed. Moreover, bactericidal activity was demonstrated to a strain elaborating the immunising carbohydrate antigen and crucially to wild-type cells of serotypes 1 and 2, thus further supporting the consideration of inner core LPS as a potential vaccine antigen to combat disease caused by Mannheimia haemolytica . Content Type Journal Article Pages 397-410 DOI 10.1007/s10719-011-9339-0 Authors Frank St. Michael, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Chantelle Cairns, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Amy Lea Filion, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Dhamodharan Neelamegan, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Suzanne Lacelle, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Andrew D. Cox, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 6

Description:    Efficient generation of useful monoclonal antibodies (mAbs) with high performance in cancer therapeutics has been expected. Generation of mAbs reactive with globotriaosylceramide (Gb3/CD77) was compared between A/J mice and Gb3/CD77 synthase-deficient (A4GalT-knockout) mice by immunizing Gb3-liposome. Specificity and functions of established antibodies were examined by ELISA, TLC- immunostaining, cytotoxicity of cancer cells and immunoblotting. Compared with results with conventional mice, better generation of mAbs with higher functions has been achieved with A4GalT-knockout mice, i.e. acquisition of IgG class antibodies, activities in antibody-dependent cell-mediated cytotoxicity, complement-dependent cytotoxicity, and aggregation activity toward a Burkitt’s lymphoma line Ramos. Binding of mAb k52 induced tyrosine phosphorylation of several proteins in Ramos cells. One of the strongest phosphorylation bands turned out to be c-Cbl. Pretreatment of B cell lines with mAbs resulted in the attenuation of BCR-mimicking signaling. All these results suggested that A4GalT-knockout mice are very useful to generate mAbs against globo-series glycolipids, and that suppressive signaling pathway driven by endogenous Gb3-ligand molecules might be present in B cells. Content Type Journal Article Pages 371-384 DOI 10.1007/s10719-011-9335-4 Authors Yuji Kondo, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Noriyo Tokuda, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Keiko Furukawa, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Reiko Ando, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Makoto Uchikawa, Tokyo Blood Center, Japanese Red Cross, 4-1-31 Hiroo, Shibuya-ku, Tokyo, 150–0012 Japan Qing Zhang, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Fan Xiaoyan, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Koichi Furukawa, Department of Biochemistry II, Nagoya University Graduate School of Medicine, 65 Tsurumai, Showa-ku, Nagoya 466–0065, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 6

Description:    The oligosaccharide structures of the structural subunit HtH1 of Haliotis tuberculata hemocyanin (HtH) were studied by mass spectral sequence analysis of the glycans. The proposed structures are based on MALDI-TOF-MS data before and after treatment with the specific exoglycosidases β1-3,4,6-galactosidase and α 1-6(〉2,3,4) fucosidase followed by sequence analysis via electrospray ionization MS/MS-spectra. In total, 15 glycans were identified as a highly heterogeneous group of structures. As in most molluscan hemocyanins, the glycans of HtH1 contain a terminal MeHex, but more interestingly, a novel structural motif was observed: MeHex[Fuc( α 1-3)-]GlcNAc, including thus MeHex and ( α 1-3)-Fuc residues being linked to an internal GlcNAc residue. While the functional unit (FU) c (HtH1-c) is completely lacking any potential glycosylation site, FU-h possesses a second exposed sugar attachment site between beta-strands 8 and 9 within the beta sandwich domain compared to the other FUs. The glycosylation pattern/sites show a high degree of conservation. In FU-h two prominent potential glycosylation sites can be detected. The finding that HtH1 is not able to form multidecameric structures in vivo could be explained by the presence of the exposed glycan on the surface of FU-h. Content Type Journal Article Pages 385-395 DOI 10.1007/s10719-011-9337-2 Authors Lyudmila Velkova, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 9 G. Bonchev St., Sofia, 1113 Bulgaria Pavlina Dolashka, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 9 G. Bonchev St., Sofia, 1113 Bulgaria Bernhard Lieb, Institute of Zoology, University of Mainz, Mϋllerweg 6, Mainz, 55099 Germany Aleksander Dolashki, Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of Sciences, 9 G. Bonchev St., Sofia, 1113 Bulgaria Wolfgang Voelter, Interfacultary Institute of Biochemistry, University of Tϋbingen, Hoppe-Seyler-Strasse 4, D-72076 Tϋbingen, Germany Jozef Van Beeumen, Laboratory of Protein Biochemistry and Biomolecular Engineering, Ghent University, KL Ledeganckstraat 35, 9000 Ghent, Belgium Bart Devreese, Laboratory of Protein Biochemistry and Biomolecular Engineering, Ghent University, KL Ledeganckstraat 35, 9000 Ghent, Belgium Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 6

Description: Erratum to: N -Glycosylation profiling of recombinant mouse extracellular superoxide dismutase produced in Chinese hamster ovary cells Content Type Journal Article Category Erratum Pages 437-437 DOI 10.1007/s10719-011-9338-1 Authors Hiroaki Korekane, Department of Disease Glycomics (Seikagaku Corporation), The Institute of Scientific and Industrial Research, Osaka University, 8–1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan Atsuko Korekane, Laboratory of Biochemistry, School of Pharmacy, Hyogo University of Health Sciences, 1-3-6 Minatojima, Chuo-ku, Kobe, Hyogo 650-8530, Japan Yoshiki Yamaguchi, Systems Glycobiology Research Group, Chemical Biology Department, Advanced Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Masaki Kato, Systems Glycobiology Research Group, Chemical Biology Department, Advanced Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan Yasuhide Miyamoto, Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511, Japan Akio Matsumoto, Department of Pharmacology, Chiba University Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan Tomoko Hasegawa, Department of Disease Glycomics (Seikagaku Corporation), The Institute of Scientific and Industrial Research, Osaka University, 8–1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan Keiichiro Suzuki, Department of Biochemistry, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan Naoyuki Taniguchi, Department of Disease Glycomics (Seikagaku Corporation), The Institute of Scientific and Industrial Research, Osaka University, 8–1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan Tomomi Ookawara, Laboratory of Biochemistry, School of Pharmacy, Hyogo University of Health Sciences, 1-3-6 Minatojima, Chuo-ku, Kobe, Hyogo 650-8530, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 6

Description:    Heparin (HP) inhibits the growth of several cell types in vitro including bovine pulmonary artery (BPA) smooth muscle cells (SMCs). In initial studies we discovered that an O -hexanoylated low-molecular-weight (LMW) HP derivative having acyl groups with 6-carbon chain length was more potent inhibitor of BPA-SMCs than the starting HP. We prepared several O -acylated LMWHP derivatives having 4-, 6-, 8-, 10-, 12-, and 18- carbon acyl chain lengths to determine the optimal acyl chain length for maximum anti-proliferative properties of BPA-SMCs. The starting LMWHP was prepared from unfractionated HP by sodium periodate treatment followed by sodium borohydride reduction. The tri- n -butylammonium salt of this LMWHP was O -acylated with butanoic, hexanoic, octanoic, decanoic, dodecanoic, and stearyl anhydrides separately to give respective O -acylated LMWHP derivatives. Gradient polyacrylamide gel electrophoresis (PAGE) was used to examine the average molecular weights of those O -acylated LMWHP derivatives. NMR analysis indicated the presence of one O -acyl group per disaccharide residue. Measurement of the inhibition of BPA-SMCS as a function of O -acyl chain length shows two optima, at a carbon chain length of 6 ( O -hexanoylated LMWHP) and at a carbon chain length 12-18 ( O -dodecanoyl and O -stearyl LMWHPs). A solution competition SPR study was performed to test the ability of different O -acylated LMWHP derivatives to inhibit fibroblast growth factor (FGF) 1 and FGF2 binding to surface-immobilized heparin. All the LMWHP derivatives bound to FGF1 and FGF2 but each exhibited slightly different binding affinity. Content Type Journal Article Pages 419-426 DOI 10.1007/s10719-011-9341-6 Authors Hari G. Garg, Department of Medicine, Pulmonary/Critical Care Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA Hicham Mrabat, Department of Medicine, Pulmonary/Critical Care Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA Lunyin Yu, Department of Medicine, Pulmonary/Critical Care Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA Charles A. Hales, Department of Medicine, Pulmonary/Critical Care Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA Boyangzi Li, Departments of Chemistry and Chemical Biology, Biology, Chemical and Biological Engineering, Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA Casey N. Moore, Departments of Chemistry and Chemical Biology, Biology, Chemical and Biological Engineering, Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA Fuming Zhang, Departments of Chemistry and Chemical Biology, Biology, Chemical and Biological Engineering, Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA Robert J. Linhardt, Departments of Chemistry and Chemical Biology, Biology, Chemical and Biological Engineering, Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 6

Description:    The tether employed to covalently attach β-mannan disaccharide glycoconjugates influences the specificity of rabbit antibodies that protect against Candida albicans . Two glycoconjugates containing (1 → 2)-β-mannan disaccharides linked to chicken serum albumin (CSA) either via a structurally uniform or via a stereodiversified spacer were prepared and evaluated in immunization trials in mice and rabbits. Immunization with conjugate vaccine possessing a structurally diversified linker induced higher IgG titers against Candida albicans cell wall phosphomannan than a conjugate with a structurally uniform linker. These results suggest that affinity maturation and the specific antibody response can be shifted towards recognition of the desired hapten by employing a linker with diversified configuration. Content Type Journal Article Pages 149-164 DOI 10.1007/s10719-011-9331-8 Authors Tomasz Lipinski, Alberta Ingenuity Centre for Carbohydrate Science, Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada Thanh Luu, Alberta Ingenuity Centre for Carbohydrate Science, Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada Pavel I. Kitov, Alberta Ingenuity Centre for Carbohydrate Science, Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada Adam Szpacenko, Alberta Ingenuity Centre for Carbohydrate Science, Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada David R. Bundle, Alberta Ingenuity Centre for Carbohydrate Science, Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 3-4

Description: Obituary: Jean Montreuil (1920–2010) Content Type Journal Article Pages 111-112 DOI 10.1007/s10719-011-9336-3 Authors Jean Claude Michakski, CNRS, UMR-8576, Structural and Functional Glycobiology, Université de Lille 1, UGSF, Bat C9, 59655 Villeneuve d’Ascq, France Hans Vliegenthart, Bijvoet Center, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 3-4

Description:    We investigated the conservation and antibody accessibility of inner core epitopes of Moraxella catarrhalis lipopolysaccharide (LPS) in order to assess their potential as vaccine candidates. Two LPS mutants, a single mutant designated lgt2 and a double mutant termed lgt2 / lgt4 , elaborating truncated inner core structures were generated in order to preclude expression of host-like outer core structures and to create an inner core structure that was shared by all three serotypes A, B and C of M. catarrhalis . Murine monoclonal antibodies (mAbs), designated MC2-1 and MC2-10 were obtained by immunising mice with the lgt2 mutant of M. catarrhalis serotype A strain. We showed that mAb MC2-1 can bind to the core LPS of wild-type (wt) serotype A, B and C organisms and concluded that mAb MC2-1 defines an immunogenic inner core epitope of M. catarrhalis LPS. We were unsuccessful in obtaining mAbs to the lgt2 / lgt4 mutant. MAb MC2-10 only recognised the lgt2 mutant and the wt serotype A strain, and exhibited a strong requirement for the terminal N -acetyl-glucosamine residue of the lgt2 mutant core oligosaccharide, suggesting that this residue was immunodominant. Subsequently, we showed that both mAbs MC2-1 and MC2-10 could facilitate bactericidal killing of the lgt2 mutant, however neither mAb could facilitate bactericidal killing of the wt serotype A strain. We then confirmed and extended the candidacy of the inner core LPS by demonstrating that it is possible to elicit functional antibodies against M. catarrhalis wt strains following immunisation of rabbits with glycoconjugates elaborating the conserved inner core LPS antigen. The present study describes three conjugation strategies that either uses amidases produced by Dictyostelium discoideum , targeting the amino functionality created by the amidase activity as the attachment point on the LPS molecule, or a strong base treatment to remove all fatty acids from the LPS, thus creating amino functionalities in the lipid A region to conjugate via maleimide-thiol linker strategies targeting the carboxyl residues of the carrier protein and the free amino functionalities of the derived lipid A region of the carbohydrate resulted in a high loading of carbohydrates per carrier protein from these carbohydrate preparations. Immunisation derived antisera from rabbits recognised fully extended M. catarrhalis LPS and whole cells. Moreover, bactericidal activity was demonstrated to both the immunising carbohydrate antigen and importantly to wt cells, thus further supporting the consideration of inner core LPS as a potential vaccine antigen to combat disease caused by M. catarrhalis . Content Type Journal Article Pages 165-182 DOI 10.1007/s10719-011-9332-7 Authors Andrew D. Cox, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Frank St. Michael, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Chantelle M. Cairns, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Suzanne Lacelle, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Amy Lea Filion, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Dhamodharan Neelamegan, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Cory Q. Wenzel, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Heather Horan, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada James C. Richards, Institute for Biological Sciences, National Research Council, 100, Sussex Drive, Ottawa, ON K1A 0R6, Canada Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 3-4

Description:    Neuroblastoma is the most common extracranial solid tumor in children and tumor ganglioside composition has been linked to its biological and clinical behavior. We recently found that high expression of complex gangliosides that are products of the enzyme GM1a/GD1b synthase predicts a more favorable outcome in human neuroblastoma, and others have shown that complex gangliosides such as GD1a inhibit metastasis of murine tumors. To determine how a switch from structurally simple to structurally complex ganglioside expression affects neuroblastoma cell behavior, we engineered IMR32 human neuroblastoma cells, which contain almost exclusively (89%) the simple gangliosides (SG) GM2, GD2, GM3, and GD3, to overexpress the complex gangliosides (CG) GM1, GD1a, GD1b and GT1b, by stable retroviral-mediated transduction of the cDNA encoding GM1a/GD1b synthase. This strikingly altered cellular ganglioside composition without affecting total ganglioside content: There was a 23-fold increase in the ratio of complex to simple gangliosides in GM1a/GD1b synthase-transduced cells (IMR32-CG) vs . wild type (IMR32) or vector-transfected (IMR32-V) cells with essentially no expression of the clinical neuroblastoma marker, GD2, confirming effectiveness of this molecular switch from simple to complex ganglioside synthesis. Probing for consequences of the switch, we found that among functional properties of IMR32-CG cells, cell migration was inhibited and Rho/Rac1 activities were altered, while proliferation kinetics and cell differentiation were unaffected. These findings further implicate cellular ganglioside composition in determining cell migration characteristics of tumor cells. This IMR32 model system should be useful in delineating the impact of ganglioside composition on tumor cell function. Content Type Journal Article Pages 137-147 DOI 10.1007/s10719-011-9330-9 Authors Lixian Dong, Center for Cancer and Immunology Research, Children’s National Medical Center, 111 Michigan Avenue, NW, Washington, DC 20010, USA Yihui Liu, Center for Cancer and Immunology Research, Children’s National Medical Center, 111 Michigan Avenue, NW, Washington, DC 20010, USA Anamaris M. Colberg-Poley, Center for Cancer and Immunology Research, Children’s National Medical Center, 111 Michigan Avenue, NW, Washington, DC 20010, USA Karen Kaucic, Center for Cancer and Immunology Research, Children’s National Medical Center, 111 Michigan Avenue, NW, Washington, DC 20010, USA Stephan Ladisch, Center for Cancer and Immunology Research, Children’s National Medical Center, 111 Michigan Avenue, NW, Washington, DC 20010, USA Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 3-4

Description:    We investigated the inhibitory activity of glycosaminoglycans (GAGs) in terms of growth, adhesion, and VacA vacuolation of Helicobacter pylori . Intact acharan sulfate (AS, MW:114 kDa) potently inhibited H. pylori adhesion to Kato III cells with IC 50 value of 1.4 mg/mL, while other GAGs did not show any inhibitory activity except for heparin which is a well-known inhibitor of H. pylori adhesion. To investigate whether low molecular weight acharan sulfate (LMWAS) can inhibit H. pylori adhesion, we performed chemical depolymerization of AS by radical reactions to obtain LMWAS. Its physicochemical properties were characterized by high-performance size exclusion chromatography (HPSEC), agarose gel electrophoresis, disaccharide compositional analysis after digestion with heparinase II, and 1 H-NMR spectroscopy. The most potent molecular size of LMWAS was 3 kDa with IC 50 value of 32 μg/mL, which is 44-fold more potent than intact AS. These results suggest that AS as well as other GAGs can be chemically depolymerized by free radicals and LMWAS compared to intact AS can be applied as a pharmaceutical candidate in order to inhibit H. pylori adhesion to Kato III cells. Content Type Journal Article Pages 411-418 DOI 10.1007/s10719-011-9340-7 Authors Joon-Soo Sim, Department of Agricultural Bio-Resources, National Academy of Agricultural Science, RDA, Suwon, 441-707 South Korea Bum-Soo Hahn, Department of Agricultural Bio-Resources, National Academy of Agricultural Science, RDA, Suwon, 441-707 South Korea A-Rang Im, College of Pharmacy, Natural Products Research Institute, Seoul National University, 599 Gwanangno, Gwanak-gu, Seoul, 151–742 South Korea Youmie Park, College of Pharmacy, Inje University, Gimhae, Gyeongnam 621–749, South Korea Ji-Eun Shin, College of Pharmacy, Kyung Hee University, Seoul, 130–701 South Korea Eun-Ah Bae, College of Pharmacy, Kyung Hee University, Seoul, 130–701 South Korea Dong-Hyun Kim, College of Pharmacy, Kyung Hee University, Seoul, 130–701 South Korea Yeong Shik Kim, College of Pharmacy, Natural Products Research Institute, Seoul National University, 599 Gwanangno, Gwanak-gu, Seoul, 151–742 South Korea Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 6

Description:    Ovomucin is a bioactive egg white glycoprotein responsible for the gel properties of fresh egg white and is believed to be involved in egg white thinning, a natural process that occurs during storage. Ovomucin is composed of two subunits: a carbohydrate-rich β-ovomucin with molecular weight of 400–610 KDa and a carbohydrate-poor α-ovomucin with molecular mass of 254 KDa. In addition to limited information on O-linked glycans of ovomucin, there is no study on either the N-glycan structures or the N-glycosylation sites. The purpose of the present study was to characterize the N-glycosylation of ovomucin from fresh eggs using nano LC ESI-MS, MS/MS and MALDI MS. Our results showed the presence of N-linked glycans on both glycoproteins. We found 18 potential N-glycosylation sites in α-ovomucin. 15 sites were glycosylated, one site was found in both glycosylated and non-glycosylated forms and two potential glycosylation sites were found unoccupied. The N-glycans of α-ovomucin found on the glycosylation sites are complex-type structures with bisecting N -acetylglucosamine. MALDI MS of the N-glycans released from α-ovomucin by PNGase F revealed that the most abundant glycan structure is a bisected type of composition GlcNAc 6 Man 3 . Two N-glycosylated sites were found in β-ovomucin. Content Type Journal Article Pages 113-123 DOI 10.1007/s10719-011-9328-3 Authors Marina Offengenden, Department of Agricultural, Food and Nutritional Science, University of Alberta, 4–10 Ag/For Centre, Edmonton, Alberta T6G 2P5, Canada Messele A. Fentabil, Department of Agricultural, Food and Nutritional Science, University of Alberta, 4–10 Ag/For Centre, Edmonton, Alberta T6G 2P5, Canada Jianping Wu, Department of Agricultural, Food and Nutritional Science, University of Alberta, 4–10 Ag/For Centre, Edmonton, Alberta T6G 2P5, Canada Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 3-4

Description:    Sulfatide is a major component of glycosphingolipids in lipoproteins. Recently, we reported that a low serum level of sulfatide in hemodialysis patients might be related to the high incidence of cardiovascular diseases. However, the serum kinetics of sulfatide in kidney disease patients and the function of endogenous serum sulfatide are still unclear. To obtain novel knowledge concerning these issues, we investigated the serum kinetics of sulfatide in 5 adult kidney transplant recipients. We also analyzed the correlated factors influencing the serum sulfatide level, using multiple regression analysis. Kidney transplantation caused a dramatic increase of serum sulfatide without an alteration of its composition in all recipients in a time-dependent manner; however, the recovery speed was slower than that of the improvement of kidney function and the serum sulfatide reached a nearly normal level after 1 year. Multiple regression analysis showed that the significant correlated factor influencing the serum sulfatide level was log duration (time parameter) throughout the observation period, and the correlated factors detected in the stable phase were the decrease of serum concentration of malondialdehyde (an oxidative stress marker) as well as the elevation of platelet count. The current study results demonstrated the gradual but reliable recovery of the serum sulfatide level in kidney transplant recipients for the first time, suggesting a close correlation between serum sulfatide and kidney function. The recovery of serum sulfatide might derive from the attenuation of systemic oxidative stress. The normal level of serum sulfatide in kidney transplant recipients might affect platelet function, and contribute to the reduction of cardiovascular disease incidence. Content Type Journal Article Pages 125-135 DOI 10.1007/s10719-011-9329-2 Authors Lixuan Wang, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Yuji Kamijo, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Akihiro Matsumoto, Department of Hepatology Internal Medicine, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Takero Nakajima, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Makoto Higuchi, Department of Nephrology Internal Medicine, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Reiji Kannagi, Research Complex for the Medicine Frontiers, Aichi Medical University, 21 Karimata, Yazako, Nagakute, Aichi 480-1195, Japan Mamoru Kyogashima, Division of Molecular Pathology, Aichi Cancer Center Research Institute, 1-1 Kanokoden, Chikusa-ku, Nagoya 464-8681, Japan Toshifumi Aoyama, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Atsushi Hara, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano 390-8621, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Numbers 3-4

Description:    Inflammation of stomach mucosa has been postulated as initiator of gastric carcinogenesis and the presence of pro-inflammatory cytokines can regulate specific genes involved in this process. The cellular expression pattern of glycosyltransferases and Lewis antigens detected in the normal mucosa changed during the neoplassic transformation. The aim of this work was to determine the regulation of specific fucosyltransferases and sialyltransferases by IL-1β and IL-6 pro-inflammatory cytokines in MKN45 gastric cancer cells. IL-1β induced significant increases in the mRNA levels of FUT1, FUT2 and FUT4, and decreases of FUT3 and FUT5. In IL-6 treatments, enhanced FUT1 and lower FUT3 and FUT5 mRNA expression were detected. No substantial changes were observed in the levels of ST3GalIII and ST3GalIV. The activation of FUT1, FUT2 and FUT4 by IL-1β is through the NF-κB pathway and the down-regulation of FUT3 and FUT5 by IL-6 is through the gp130/STAT-3 pathway, since they are inhibited specifically by panepoxydone and AG490, respectively. The levels of Lewis antigens after IL-1β or IL-6 stimulation decreased for sialyl-Lewis x, and no significant differences were found in the rest of the Lewis antigens analyzed, as it was also observed in subcutaneous mice tumors from MKN45 cells treated with IL-1β or IL-6. In addition, in 61 human intestinal-type gastric tumors, sialyl-Lewis x was highly detected in samples from patients that developed metastasis. These results indicate that the expression of the fucosyltransferases involved in the synthesis of Lewis antigens in gastric cancer cells can be specifically modulated by IL-1β and IL-6 inflammatory cytokines. Content Type Journal Article Pages 99-110 DOI 10.1007/s10719-011-9327-4 Authors Mercè Padró, Programa de Recerca en Càncer, IMIM-Hospital del Mar, 08003 Barcelona, Spain Raquel Mejías-Luque, Programa de Recerca en Càncer, IMIM-Hospital del Mar, 08003 Barcelona, Spain Lara Cobler, Programa de Recerca en Càncer, IMIM-Hospital del Mar, 08003 Barcelona, Spain Marta Garrido, Programa de Recerca en Càncer, IMIM-Hospital del Mar, 08003 Barcelona, Spain Marta Pérez-Garay, Unitat de Bioquimica i Biologia Molecular, Departament de Biologia, Universitat de Girona, 17071 Girona, Spain Sònia Puig, Programa de Recerca en Càncer, IMIM-Hospital del Mar, 08003 Barcelona, Spain Rosa Peracaula, Unitat de Bioquimica i Biologia Molecular, Departament de Biologia, Universitat de Girona, 17071 Girona, Spain Carme de Bolós, Programa de Recerca en Càncer, IMIM-Hospital del Mar, 08003 Barcelona, Spain Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 2

Description:    Free ceramides and glycosphingolipids (GSLs) are important components of the membrane microdomain and play significant roles in cell survival. Recent studies have revealed that both fatty acids and long-chain bases (LCBs) are more diverse than expected, in terms of i) alkyl chain length, ii) hydroxylation and iii) the presence or absence of double bonds. Electrospray ionization mass spectrometry and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) have been well utilized to characterize sphingolipids with high throughput, but reports to date have not fully characterized various types of ceramide species such as hydroxyl fatty acids and/or trihydroxy-LCBs of both free ceramides and the constituent ceramides in neutral GSLs. We performed a systematic analysis of both ceramide species, including LCBs with nona-octadeca lengths using MALDI-TOF MS with high-energy collision-induced dissociation (CID) at 20 keV. Using both protonated and sodiated ions, this technique enabled us to propose general rules to discriminate between isomeric and isobaric ceramide species, unrelated to the presence or absence of sugar chains. In addition, this high-energy CID generated 3,5 A ions, indicating Hex1-4Hex linkage in the sugar chains. Using this method, we demonstrated distinct differences among ceramide species, including free ceramides, sphingomyelins, and neutral GSLs of glucosylceramides, galactosylceramides, lactosylceramides, globotriaosylceramides and Forssman glycolipids in the equine kidneys. Content Type Journal Article Pages 67-87 DOI 10.1007/s10719-011-9325-6 Authors Kouji Tanaka, Division of Molecular Pathology, Aichi Cancer Center Research Institute, Nagoya, Aichi 464-8681, Japan Masaki Yamada, Shimadzu Corporation, Kyoto, 604-8511 Japan Keiko Tamiya-Koizumi, Division of Molecular Pathology, Aichi Cancer Center Research Institute, Nagoya, Aichi 464-8681, Japan Reiji Kannagi, Division of Molecular Pathology, Aichi Cancer Center Research Institute, Nagoya, Aichi 464-8681, Japan Toshifumi Aoyama, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, Matsumoto, Nagano 390-8621, Japan Atsushi Hara, Department of Metabolic Regulation, Institute on Aging and Adaptation, Shinshu University Graduate School of Medicine, Matsumoto, Nagano 390-8621, Japan Mamoru Kyogashima, Division of Molecular Pathology, Aichi Cancer Center Research Institute, Nagoya, Aichi 464-8681, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 28 Journal Issue Volume 28, Number 2

Description:    During the biosynthesis of N-glycans in multicellular eukaryotes, glycans with the compositions Man 5 GlcNAc 2-3 are key intermediates. However, to reach this ‘decision point’, these N-glycans are first processed from Glc 3 Man 9 GlcNAc 2 through to Man 5 GlcNAc 2 by a number of glycosidases, whereby up to four α1-2-linked mannose residues are removed by class I mannosidases (glycohydrolase family 47). Whereas in the yeast Saccharomyces cerevisiae there are maximally three members of this protein family, in higher organisms there are multiple class I mannosidases residing in the endoplasmic reticulum and Golgi apparatus. The genome of the model nematode Caenorhabditis elegans encodes seven members of this protein family, whereby four are predicted to be classical processing mannosidases and three are related proteins with roles in quality control. In this study, cDNAs encoding the four predicted mannosidases were cloned and expressed in Pichia pastoris and the activity of these enzymes, designated MANS-1, MANS-2, MANS-3 and MANS-4, was verified. The first two can, dependent on the incubation time, remove three to four residues from Man 9 GlcNAc 2 , whereas the action of the other two results in the appearance of the B isomer of Man 8 GlcNAc 2 ; together the complementary activities of these enzymes result in processing to Man 5 GlcNAc 2 . With these data, another gap is closed in our understanding of the N-glycan biosynthesis pathway of the nematode worm. Content Type Journal Article Pages 1-7 DOI 10.1007/s10719-012-9378-1 Authors Iain B. H. Wilson, Department für Chemie, Universität für Bodenkultur, Muthgasse 18, 1190 Wien, Austria Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Mass spectrometry plays an increasingly important role in structural glycomics. This review provides an overview on currently used mass spectrometric approaches such as the characterization of glycans, the analysis of glycopeptides obtained by proteolytic cleavage of proteins and the analysis of glycosphingolipids. The given examples are demonstrating the application of mass spectrometry to study glycosylation changes associated with congenital disorders of glycosylation, lysosomal storage diseases, autoimmune diseases and cancer. Content Type Journal Article Pages 1-12 DOI 10.1007/s10719-012-9376-3 Authors Manfred Wuhrer, Department of Parasitology, Biomolecular Mass Spectrometry Unit, Leiden University Medical Center, Albinusdreef 2, 2333ZA Leiden, The Netherlands Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080

Description:    Intercellular adhesion molecule-5 (ICAM-5, telencephalin) is a dendritically polarized type I membrane glycoprotein, and promotes dendritic filopodia formation. Although we have determined the N-glycan structures of ICAM-5 in a previous report, their function is unknown. Here, we produced fifteen ICAM-5 gene constructs, in which each potential N-glycosylation site was mutated, to elucidate the function of the N-glycans of ICAM-5, and observed the effects of transfection of them on a neuronal cell line, Neuro-2a (N2a). Only the N54Q mutant, which is the mutant for the most N-terminal glycosylation site, failed to induce filopodia-like protrusions in N2a cells. Immunofluorescence staining and cell surface biotinylation revealed that N54Q ICAM-5 was confined to the ER and also could not be expressed on the cell surface. This is further supported by the biochemical evidence that almost all N-glycans of N54Q ICAM-5 were digested by Endo glycosidase H and peptide:N-glycanase, indicating that almost all of them retain high-mannose-type structures in ER. In additon, it also failed to form disulfide bonds or functional protein complexes. The stable transformants of N54Q ICAM-5 showed retarded cell growth, but it was interesting that there was no apparent ER stress, because the mutant was sequentially degraded via ER associated degradation pathway by comparing the susceptibilities of the responses to various inhibitors of this pathway in wild-type and N54Q ICAM-5 transfectants. Taken together, the Asn 54 -linked glycan is necessary for normal trafficking and function of ICAM-5, but is unassociated with ER-associated degradation of it. Content Type Journal Article Pages 47-55 DOI 10.1007/s10719-011-9363-0 Authors Tomohiro Ohgomori, Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan Tomohisa Nanao, Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan Akinori Morita, Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan Masahiko Ikekita, Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan Journal Glycoconjugate Journal Online ISSN 1573-4986 Print ISSN 0282-0080 Journal Volume Volume 29 Journal Issue Volume 29, Number 1