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Arne O Melleby, Mari E Strand, Andreas Romaine, Kate M Herum, Biljana Skrbic, Christen P Dahl, Ivar Sjaastad, Arnt E Fiane, Jorge Filmus, Geir Christensen, Ida G Lunde
Pressure overload is a frequent cause of heart failure. Heart failure affects millions of patients worldwide and is a major cause of morbidity and mortality. Cell surface proteoglycans are emerging as molecular players in cardiac remodeling, and increased knowledge about their regulation and function is needed for improved understanding of cardiac pathogenesis. Here we investigated glypicans (GPC1-6), a family of evolutionary conserved heparan sulfate proteoglycans anchored to the extracellular leaflet of the cell membrane, in experimental and clinical heart failure, and explored the function of glypican-6 in cardiac cells in vitro...
2016: PloS One
Pavel Damborský, Katarzyna M Koczula, Andrea Gallotta, Jaroslav Katrlík
Lateral flow assays (LFAs) enable the simple and rapid detection and quantification of analytes and is popular for point-of-care (PoC), point-of-use and outdoor testing applications. LFAs typically depend on antibody or nucleic acid based recognition. We present the innovative concept of a LFA using lectins in the role of the biorecognition element. Lectins are a special kind of glycan-binding protein and the lectin-based LFA herein described was developed for the determination of the glycosylation of free prostate specific antigen (PSA)...
October 21, 2016: Analyst
Emily L Clark, Madhu Emmadi, Katharine L Krupp, Ananda R Podilapu, Jennifer D Helble, Suvarn S Kulkarni, Danielle H Dube
Bacterial glycans contain rare, exclusively bacterial monosaccharides that are frequently linked to pathogenesis and essentially absent from human cells. Therefore, bacterial glycans are intriguing molecular targets. However, systematic discovery of bacterial glycoproteins is hampered by the presence of rare deoxy amino sugars, which are refractory to traditional glycan-binding reagents. Thus, the development of chemical tools that label bacterial glycans is a crucial step toward discovering and targeting these biomolecules...
October 21, 2016: ACS Chemical Biology
Y Sun, Y X Wang, J Zhang, X W Yu, S S Ge, H Wang, J Dong, X H Guo, W Wang
Objective: To analyze and compare the gender specific structure profiles of immunoglobulin G(IgG) N-glycans and provide evidence to understand the distribution of the structure of IgG N-glycans in general population. Methods: A cluster sampling was conducted to randomly select 669 participants from Beijing Xuanwu Hospital during January-June 2012. Venous blood sample (5 ml) was collected from each participant with vacuum negative pressure tube containing EDTA. Plasma IgG N-glycome was determined using ultra-performance liquid chromatography (UPLC) on Waters BEH Glycan chromatography column and mass spectrometry after the separation, purification and labeling...
October 10, 2016: Zhonghua Liu Xing Bing Xue za Zhi, Zhonghua Liuxingbingxue Zazhi
Tatiana Pochechueva, Alexander Chinarev, Andreas Schoetzau, André Fedier, Nicolai V Bovin, Neville F Hacker, Francis Jacob, Viola Heinzelmann-Schwarz
Altered levels of naturally occurring anti-glycan antibodies (AGA) circulating in human blood plasma are found in different pathologies including cancer. Here the levels of AGA directed against 22 negatively charged (sialylated and sulfated) glycans were assessed in high-grade serous ovarian cancer (HGSOC, n = 22) patients and benign controls (n = 31) using our previously developed suspension glycan array (SGA). Specifically, the ability of AGA to differentiate between controls and HGSOC, the most common and aggressive type of ovarian cancer with a poor outcome was determined...
2016: PloS One
Christopher J Day, Rebecca M King, Lucy K Shewell, Greg Tram, Tahria Najnin, Lauren E Hartley-Tassell, Jennifer C Wilson, Aaron D Fleetwood, Igor B Zhulin, Victoria Korolik
A rare chemotaxis receptor, Tlp11, has been previously identified in invasive strains of Campylobacter jejuni, the most prevalent cause of bacterial gastroenteritis worldwide. Here we use glycan and small-molecule arrays, as well as surface plasmon resonance, to show that Tlp11 specifically interacts with galactose. Tlp11 is required for the chemotactic response of C. jejuni to galactose, as shown using wild type, allelic inactivation and addition mutants. The inactivated mutant displays reduced virulence in vivo, in a model of chicken colonization...
October 20, 2016: Nature Communications
Man Kyu Shim, Hong Yeol Yoon, Ju Hee Ryu, Heebeom Koo, Sangmin Lee, Jae Hyung Park, Jong-Ho Kim, Seulki Lee, Martin G Pomper, Ick Chan Kwon, Kwangmeyung Kim
Recently, metabolic glycoengineering with bioorthogonal click reactions has focused on improving the tumor targeting efficiency of nanoparticles as delivery vehicles for anticancer drugs or imaging agents. It is the key technique for developing tumor-specific metabolic precursors that can generate unnatural glycans on the tumor-cell surface. A cathepsin B-specific cleavable substrate (KGRR) conjugated with triacetylated N-azidoacetyl-d-mannosamine (RR-S-Ac3 ManNAz) was developed to enable tumor cells to generate unnatural glycans that contain azide groups...
October 20, 2016: Angewandte Chemie
Ju Yeon Lee, Hyun Kyoung Lee, Gun Wook Park, Heeyoun Hwang, Hoi Keun Jeong, Ki Na Yun, Eun Sun Ji, Kwang Hoe Kim, Jun Seok Kim, Jong Won Kim, Sung Ho Yun, Chi-Won Choi, Seung Il Kim, Jong-Sun Lim, Seul-Ki Jeong, Young-Ki Paik, Soo-Youn Lee, Jisook Park, Su Yeon Kim, Young-Jin Choi, Yong-In Kim, Jawon Seo, Je-Yoel Cho, Myoung Jin Oh, Nari Seo, Hyun Joo An, Jin Young Kim, Jong Shin Yoo
Glycoprotein conformations are complex and heterogeneous. Currently, site-specific characterization of glycopeptides is a challenge. Herein, we sought to establish an efficient method of N-glycoprotein characterization using mass spectrometry. Using alpha-1-acid glycoprotein (AGP) as a model N-glycoprotein, its tryptic N-glycopeptides were identified, and the reproducibility of the data was examined in seven laboratories running different liquid chromatography-mass tandem spectrometry (LC-MS/MS) platforms. We used three test samples and one blind sample to evaluate instrument performance with entire sample preparation workflow...
October 19, 2016: Journal of Proteome Research
Nathan J Hare, Ling Y Lee, Ian Loke, Warwick J Britton, Bernadette M Saunders, Morten Thaysen-Andersen
Tuberculosis (TB) remains a prevalent and lethal infectious disease. The glycobiology associated with Mycobacterium tuberculosis infection of front-line alveolar macrophages is still unresolved. Herein, we investigated the regulation of protein N-glycosylation in human macrophages and their secreted microparticles (MPs) used for intercellular communication upon M. tb infection. LC-MS/MS-based proteomics and glycomics were performed to monitor the regulation of glycosylation enzymes and receptors and the N-glycome in in vitro-differentiated macrophages and in isolated MPs upon M...
October 19, 2016: Journal of Proteome Research
Yoshiyuki Goto, Satoshi Uematsu, Hiroshi Kiyono
Intestinal epithelial cells apically express glycans, especially α1,2-fucosyl linkages, which work as a biological interface for the host-microbe interaction. Emerging studies have shown that epithelial α1,2-fucosylation is regulated by microbes and by group 3 innate lymphoid cells (ILC3s). Dysregulation of the gene (FUT2) encoding fucosyltransferase 2, an enzyme governing epithelial α1,2-fucosylation, is associated with various human disorders, including infection and chronic inflammatory diseases. This suggests a critical role for an interaction between microbes, epithelial cells and ILC3s mediated via glycan residues...
October 19, 2016: Nature Immunology
Behjatolah Monzavi-Karbassi, Fariba Jousheghany, Thomas Kieber-Emmons
Development of cancer vaccines targeting tumor-associated antigens (TAAs) is an alternative approach to chemotherapy with sustained anti-tumor effects. The success of active immunotherapy has been hampered by tumor-induced immune suppressors. Regulatory T cells (Tregs) are a population of immune suppressors with a proven role in regulating anti-tumor immune responses. Removing or subduing Tregs activity leads to more robust anti-tumor immune responses. Here, we used a cell-based vaccination strategy in the 4T1 murine mammary model to examine whether bulk removal of certain TAAs, using their glycan profile, can affect the immunogenicity of the vaccine...
October 19, 2016: Immunological Investigations
Alexandre Guillot, Manuel Dauchez, Nicolas Belloy, Jessica Jonquet, Laurent Duca, Beatrice Romier, Pascal Maurice, Laurent Debelle, Laurent Martiny, Vincent Durlach, Stephanie Baud, Sebastien Blaise
Sialic acids (SA) are monosaccharides that can be located at the terminal position of glycan chains on a wide range of proteins. The post-translational modifications, such as N-glycan chains, are fundamental to protein functions. Indeed, the hydrolysis of SA by specific enzymes such as neuraminidases can lead to drastic modifications of protein behavior. However, the relationship between desialylation of N-glycan chains and possible alterations of receptor function remains unexplored. Thus, the aim of the present study is to establish the impact of SA removal from N-glycan chains on their conformational behavior...
October 19, 2016: Scientific Reports
Laura Bouché, Maria Panico, Paul Hitchen, Daniel Binet, Federico Sastre, Alexandra Faulds-Pain, Esmeralda Valiente, Evgeny Vinogradov, Annie Aubry, Kelly Fulton, Susan Twine, Susan M Logan, Brendan W Wren, Anne Dell, Howard R Morris
Glycosylation of flagellins is a well recognized property of many bacterial species. In this study we describe the structural characterization of novel flagellar glycans from a number of hypervirulent strains of C. difficile. We used mass spectrometry (nano LC- MS and MS/MS analysis) to identify a number of putative glycopeptides which carried a variety of glycoform substitutions each of which was linked through an initial HexNAc residue to Ser or Thr. Detailed analysis of a LLDGSSTEIR glycopeptide released by tryptic digestion, which carried two variant structures, revealed that the glycopeptide contained, in addition to carbohydrate moieties, a novel structural entity...
October 7, 2016: Journal of Biological Chemistry
Atul Kumar, Petra Sýkorová, Gabriel Demo, Pavel Dobeš, Pavel Hyršl, Michaela Wimmerová
Photorhabdus luminescens is known for its symbiosis with the entomopathogenic nematode Heterorhabditis bacteriophora and its pathogenicity towards insect larvae. A hypothetical protein from P. luminescens was identified, purified from the native source and characterized as an L-fucose-binding lectin, named PLL. Glycan array and biochemical characterization data revealed PLL to be specific towards L-fucose and the disaccharide glycan 3,6-O-Me2-Glcβ1-4(2,3-O-Me2)Rhaα-O-(p-C6H4)-OCH2CH2NH2. PLL was discovered to be a homo-tetramer with an inter-subunit disulphide bridge...
October 7, 2016: Journal of Biological Chemistry
Shuxin Shang, Lu Sun, Wei Li, Xue Qin, Shu Zhang, Yinkun Liu
Fucosylated glycans play a critical role in cell biology processes during the occurrence and development of tumor, such as cell signal transduction, cellular differentiation, ligand-receptor interactions, and metastasis formation. Fucosylated haptoglobin (Hp) was reported to be increased in hepatocellular carcinoma (HCC). In this study, we aimed to develop a convenient and sensitive AAL@Magbeads-based Hp ELISA assay to rapidly detect fucosylated Hp in 270 serum samples from HCC patients and normal controls...
September 2016: Discovery Medicine
Dayoung Park, Narine Arabyan, Cynthia C Williams, Ting Song, Anupam Mitra, Bart C Weimer, Emanual Maverakis, Carlito B Lebrilla
Although gut host-pathogen interactions are glycan-mediated processes, few details are known about the participating structures. Here we employ high-resolution mass spectrometric profiling to comprehensively identify and quantitatively measure the exact modifications of native intestinal epithelial cell surface N-glycans induced by S. Typhimurium infection. Sixty minutes post-infection, select sialylated structures showed decreases in terms of total number and abundances. To assess the effect of cell surface mannosylation, we selectively rerouted glycan expression on the host using the alpha-mannosidase inhibitor, kifunensine, toward overexpression of high mannose...
October 17, 2016: Molecular & Cellular Proteomics: MCP
Sandra Cascio, Olivera J Finn
Altered glycosylation of mucin 1 (MUC1) on tumor cells compared to normal epithelial cells was previously identified as an important antigenic modification recognized by the immune system in the process of tumor immunosurveillance. This tumor form of MUC1 is considered a viable target for cancer immunotherapy. The importance of altered MUC1 glycosylation extends also to its role as a promoter of chronic inflammatory conditions that lead to malignant transformation and cancer progression. We review here what is known about the role of specific cancer-associated glycans on MUC1 in protein-protein interactions and intracellular signaling in cancer cells and in their adhesion to each other and the tumor stroma...
October 13, 2016: Biomolecules
Keith B Boyle, Teresa L M Thurston, Felix Randow
Defense of the mammalian cell cytosol against Salmonella invasion is reliant upon capture of the infiltrating bacteria by macroautophagy (hereafter autophagy), a process controlled by the kinase TBK1. In our recent study we showed that recruitment of TBK1 activity to Salmonella stabilizes the key autophagy regulator WIPI2 on those bacteria, a novel and essential function for TBK1 in the control of the early steps of antibacterial autophagy. Substantial redundancy exists in the precise recruitment mechanism for TBK1 because engagement with any of several Salmonella-associated 'eat-me' signals, including host-derived glycans, and K48- and K63-linked ubiquitin chains, suffices to recruit TBK1 functionality...
October 18, 2016: Autophagy
Zhigang Wu, Yunpeng Liu, Cheng Ma, Lei Li, Jing Bai, Lauren Byrd-Leotis, Yi Lasanajak, Yuxi Guo, Liuqing Wen, He Zhu, Jing Song, Yanhong Li, David A Steinhauer, David F Smith, Baohua Zhao, Xi Chen, Wanyi Guan, Peng George Wang
Glycans play diverse roles in a wide range of biological processes. Research on glycan-binding events is essential for learning their biological and pathological functions. However, the functions of terminal and internal glycan epitopes exhibited during binding with glycan-binding proteins (GBPs) and/or viruses need to be further identified. Therefore, a focused library of 36 biantennary asparagine (Asn)-linked glycans with some presenting tandem glycan epitopes was synthesized via a combined Core Isolation/Enzymatic Extension (CIEE) and one-pot multienzyme (OPME) synthetic strategy...
October 18, 2016: Organic & Biomolecular Chemistry
Carmen Jiménez-Castells, Jorick Vanbeselaere, Sonja Kohlhuber, Bärbel Ruttkowski, Anja Joachim, Katharina Paschinger
BACKGROUND: The porcine nodule worm Oesophagostomum dentatum is a strongylid class V nematode rather closely related to the model organism Caenorhabditis elegans. However, in contrast to the non-parasitic C. elegans, the parasitic O. dentatum is an obligate sexual organism, which makes both a gender and developmental glycomic comparison possible. METHODS: Different enzymatic and chemical methods were used to release N-glycans from male and female O. dentatum as well as from L3 and L4 larvae...
October 15, 2016: Biochimica et Biophysica Acta
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