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N-glycosylation

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https://www.readbyqxmd.com/read/28531887/expression-and-characterization-of-membrane-type-4-matrix-metalloproteinase-mt4-mmp-and-its-different-forms-in-melanoma
#1
Bettina Hieronimus, Julian Pfohl, Christian Busch, Lutz Graeve
BACKGROUND/AIMS: Membrane-type matrix metalloproteinases (MT-MMPs) are expressed on the cell surface and hydrolyze extracellular matrix components and signaling molecules by which they influence cancer cell migration and metastasis. Two of the six known MT-MMPs are anchored to the plasma membrane via a GPI anchor, one of which is MT4-MMP. Only little is known about MT4-MMP expression, synthesis, regulation and degradation. METHODS: We analyzed several human cancer cell lines as well as tissue homogenates using Western blotting and quantitative PCR for the expression of MT4-MMP...
May 25, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/28529241/downregulation-of-transcription-factor-sp1-suppresses-malignant-properties-of-a549-human-lung-cancer-cell-line-with-decreased-%C3%AE-4-galactosylation-of-highly-branched-n-glycans
#2
Kodai Muramoto, Riho Tange, Takayuki Ishii, Kana Miyauchi, Takeshi Sato
Dramatic changes in the glycan structures of cell surface proteins have been observed upon malignant transformation of cells as induced by the altered expression levels of glycosyltransferases. Such changes are closely associated with the malignant properties of cancer cells. Transcription factor Sp1 regulates the gene expression of various molecules including glycosyltransferases. Herein, we investigated whether or not Sp1-downregulation affects to N-glycosylation of glycoproteins and malignant properties of A549 human lung carcinoma cell line...
May 20, 2017: Biological & Pharmaceutical Bulletin
https://www.readbyqxmd.com/read/28522347/functional-analysis-of-tcctlp-5c2-a-chymotrypsin-like-serine-protease-needed-for-molting-in-tribolium-castaneum
#3
Daniel Albaum, Gunnar Broehan, Subbaratnam Muthukrishnan, Hans Merzendorfer
In a previous study, we have characterized a gene family encoding chymotrypsin-like proteases from the red flour beetle, Tribolium castaneum (TcCTLPs). We identified 14 TcCTLP genes that were predominantly expressed in the midgut, where they presumably function in digestion. Two genes (TcCTLP-6C and TcCTLP-5C2), however, additionally showed considerable expression in the carcass, and RNAi studies demonstrated that they are required for molting (Broehan et al., 2010; Insect Biochem. Mol. Biol 40, 274-83). Thus, the enzyme has distinct functions in different physiological environments...
May 15, 2017: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/28517004/exploitation-of-new-structurally-diverse-d-glucuronamide-containing-n-glycosyl-compounds-synthesis-and-anticancer-potential
#4
Nuno M Xavier, Alexandre Porcheron, Daniela Batista, Radek Jorda, Eva Řezníčková, Vladimír Kryštof, M Conceição Oliveira
The synthesis and anticancer evaluation of novel N-glycosyl derivatives containing N-substituted glucuronamide moieties, as nucleoside analogs or as prospective mimetics of glycosyl phosphates or of nucleotides, is reported. These compounds comprise N-anomerically-linked nucleobases or motifs that are surrogates of a phosphate group, such as sulfonamide or phosphoramidate moieties. 1-Sulfonamido glucuronamides containing N-benzyl, N-propargyl or N-dodecyl carboxamide units were synthesized through glycosylation of methanesulfonamide with tetra-O-acetyl glucuronamides...
May 18, 2017: Organic & Biomolecular Chemistry
https://www.readbyqxmd.com/read/28516782/exploring-site-specific-n-glycosylation-of-hek293-and-plant-produced-human-iga-isotypes
#5
Kathrin Göritzer, Daniel Maresch, Friedrich Altmann, Christian Obinger, Richard Strasser
The full potential of recombinant Immunoglobulin A as therapeutic antibody is not fully explored, owing to the fact that structure-function relationships of these extensively glycosylated proteins are not well understood. Here, monomeric IgA1, IgA2m(1) and IgA2m(2) variants of the anti-HER2 antibody (IgG1) trastuzumab were expressed in glyco-engineered Nicotiana benthamiana plants and in human HEK293-6E cells. All three IgA isotypes were purified and subjected to biophysical and biochemical characterization...
May 18, 2017: Journal of Proteome Research
https://www.readbyqxmd.com/read/28512020/molecular-epidemiology-of-porcine-reproductive-and-respiratory-syndrome-virus-in-central-china-since-2014-the-prevalence-of-nadc30-like-prrsvs
#6
Lin-Jian Wang, Weitao Xie, Xin-Xin Chen, Songlin Qiao, Mengmeng Zhao, Yu Gu, Bao-Lei Zhao, Gaiping Zhang
Porcine reproductive and respiratory syndrome (PRRS), characterized by respiratory disorders in piglets and reproductive failure in sows, is still the great threat of swine industry. Recently, Emergence of the novel NADC30-like PRRS viruses (PRRSVs) has caused widespread outbreaks of PRRS. To investigate the epidemic characteristics of PRRSVs in Central China since 2014, 6372 clinical serum samples were tested by ELISA, 250 tissue samples were tested by RT-PCR, and among these, 30 ORF5 and 17 Nsp2 genes sequences were analyzed...
May 13, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28510654/asn-linked-oligosaccharide-chain-of-a-crenarchaeon-pyrobaculum-calidifontis-is-reminiscent-of-the-eukaryotic-high-mannose-type-glycan
#7
Daisuke Fujinami, Yuya Taguchi, Daisuke Kohda
Pyrobaculum calidifontis is a hyperthermophilic archaeon that belongs to the phylum Crenarchaeota. In contrast to the phylum Euryarchaeota, only the N-glycan structure of the genus Sulfolobus is known in Crenarchaeota. Here, we enriched glycoproteins from cultured P. calidifontis cells, by ConA lectin chromatography. The MASCOT search identified proteins with at least one potential N-glycosylation site. The MS/MS analysis of twelve small tryptic glycopeptides confirmed the canonical N-glycosylation consensus in P...
May 16, 2017: Glycobiology
https://www.readbyqxmd.com/read/28501204/quantification-of-n-glycosylation-site-occupancy-status-based-on-labeling-label-free-strategies-with-lc-ms-ms
#8
REVIEW
Shu Zhang, Wei Li, Haojie Lu, Yinkun Liu
Protein N-glycosylation plays important roles in physiological and pathological processes. Characterizing the site-specific N-glycosylation including N-glycan macroheterogeneity (glycosylation site occupancy) and microheterogeneity (site-specific glycan structure) is important for understanding of glycoprotein biosynthesis and function. N-Glycan macroheterogeneity is a physiological property of glycoprotein and the technical obstacles have restricted research into the regulation and functions of this heterogeneity...
August 1, 2017: Talanta
https://www.readbyqxmd.com/read/28493133/engineer-medium-and-feed-for-modulating-n-glycosylation-of-recombinant-protein-production-in-cho-cell-culture
#9
Yuzhou Fan, Helene Faustrup Kildegaard, Mikael Rørdam Andersen
Chinese hamster ovary (CHO) cells have become the primary expression system for the production of complex recombinant proteins due to their long-term success in industrial scale production and generating appropriate protein N-glycans similar to that of humans. Control and optimization of protein N-glycosylation is crucial, as the structure of N-glycans can largely influence both biological and physicochemical properties of recombinant proteins. Protein N-glycosylation in CHO cell culture can be controlled and tuned by engineering medium, feed, culture process, as well as genetic elements of the cell...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28489325/reducing-macro-and-microheterogeneity-of-n-glycans-enabled-solving-the-crystal-structure-of-the-lectin-and-egf-like-domains-of-human-l-selectin-at-1-9-%C3%A3-resolution
#10
Stefanie Wedepohl, Jens Dernedde, Ardeschir Vahedi-Faridi, Rudolf Tauber, Wolfram Saenger, Haydar Bulut
L-selectin is a cell adhesion receptor located on the surface of most leukocytes and contains a total of 7 N-glycosylation sites. To obtain the crystal structure of human L-selectin, we expressed a shortened version comprising the C-type lectin and EGF-like domains of L-selectin (termed LE hereafter) and systematically analysed mutations of the three glycosylation sites at Asn22, Asn66, and Asn139 in order to reduce macroheterogeneity. After we further removed microheterogeneity, we obtained crystals which diffracted X-rays up to 1...
May 10, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28487369/n-glycosylation-affects-the-stability-and-barrier-function-of-the-muc16-mucin
#11
Takazumi Taniguchi, Ashley M Woodward, Paula Magnelli, Nicole M McColgan, Sylvain Lehoux, Sarah Melissa P Jacobo, Jérôme Mauris, Pablo Argüeso
Transmembrane mucins are highly O-glycosylated glycoproteins that coat the apical glycocalyx on mucosal surfaces and represent the first line of cellular defense against infection and injury. Relatively low levels of N-glycans are found on transmembrane mucins, and their structure and function remains poorly characterized. We previously reported that carbohydrate-dependent interactions of transmembrane mucins with galectin-3 contribute to maintain the epithelial barrier at the ocular surface. Now, using MALDI-TOF mass spectrometry, we report that transmembrane mucin N-glycans in differentiated human corneal epithelial cells contain primarily complex-type structures with N-acetyllactosamine, a preferred galectin ligand...
May 9, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28486782/role-of-n-glycosylation-in-egfr-ectodomain-ligand-binding
#12
Maryam Azimzadeh Irani, Srinivasaraghavan Kannan, Chandra Verma
The Epidermal Growth Factor Receptor (EGFR) is a tyrosine kinase protein, overexpressed in several cancers. The extracellular domain of EGFR is known to be heavily glycosylated. Growth factor (mostly Epidermal Growth Factor or EGF) binding activates EGFR. This occurs by inducing the transition from the autoinhibited tethered conformation to an extended conformation of the monomeric form of EGFR and by stabilizing the flexible pre-formed dimer. Activated EGFR adopts a back-to-back dimeric conformation after binding of another homologous receptor to its extracellular domain as the dimeric partner...
May 9, 2017: Proteins
https://www.readbyqxmd.com/read/28486580/enterocyte-glycosylation-is-responsive-to-changes-in-extracellular-conditions-implications-for-membrane-functions
#13
Dayoung Park, Gege Xu, Mariana Barboza, Ishita M Shah, Maurice Wong, Helen Raybould, David A Mills, Carlito B Lebrilla
Epithelial cells in the lining of the intestines play critical roles in maintaining homeostasis while challenged by dynamic and sudden changes in luminal contents. Given the high density of glycosylation that encompasses their extracellular surface, environmental changes may lead to extensive reorganization of membrane-associated glycans. However, neither the molecular details nor the consequences of conditional glycan changes are well understood. Here we assessed the sensitivity of Caco-2 and HT-29 membrane N-glycosylation to variations in (i) dietary elements; (ii) microbial fermentation products; and (ii) cell culture parameters relevant to intestinal epithelial cell growth and survival...
May 9, 2017: Glycobiology
https://www.readbyqxmd.com/read/28486471/model-based-analysis-of-n-glycosylation-in-chinese-hamster-ovary-cells
#14
Frederick J Krambeck, Sandra V Bennun, Mikael R Andersen, Michael J Betenbaugh
The Chinese hamster ovary (CHO) cell is the gold standard for manufacturing of glycosylated recombinant proteins for production of biotherapeutics. The similarity of its glycosylation patterns to the human versions enable the products of this cell line favorable pharmacokinetic properties and lower likelihood of causing immunogenic responses. Because glycan structures are the product of the concerted action of intracellular enzymes, it is difficult to predict a priori how the effects of genetic manipulations alter glycan structures of cells and therapeutic properties...
2017: PloS One
https://www.readbyqxmd.com/read/28482115/arabidopsis-thaliana-fla4-functions-as-a-glycan-stabilized-soluble-factor-via-its-carboxy-proximal-fasciclin-1-domain
#15
Hui Xue, Christiane Veit, Lindy Abas, Theodora Tryfona, Daniel Maresch, Martiniano M Ricardi, Jose M Estevez, Richard Strasser, Georg J Seifert
Fasciclin like arabinogalactan proteins (FLAs) are involved in numerous important functions in plants but the relevance of their complex structure to physiological function and cellular fate is unresolved. Using a fully functional fluorescent version of Arabidopsis thaliana FLA4 we show that this protein is localized at the plasma membrane as well as in endosomes and soluble in the apoplast. FLA4 is likely to be GPI-anchored, is highly N-glycosylated and carries two O-glycan epitopes previously associated with AGPs...
May 8, 2017: Plant Journal: for Cell and Molecular Biology
https://www.readbyqxmd.com/read/28481222/hepatic-metal-ion-transporter-zip8-regulates-manganese-homeostasis-and-manganese-dependent-enzyme-activity
#16
Wen Lin, David R Vann, Paschalis-Thomas Doulias, Tao Wang, Gavin Landesberg, Xueli Li, Emanuela Ricciotti, Rosario Scalia, Miao He, Nicholas J Hand, Daniel J Rader
Genetic variants at the solute carrier family 39 member 8 (SLC39A8) gene locus are associated with the regulation of whole-blood manganese (Mn) and multiple physiological traits. SLC39A8 encodes ZIP8, a divalent metal ion transporter best known for zinc transport. Here, we hypothesized that ZIP8 regulates Mn homeostasis and Mn-dependent enzymes to influence metabolism. We generated Slc39a8-inducible global-knockout (ZIP8-iKO) and liver-specific-knockout (ZIP8-LSKO) mice and observed markedly decreased Mn levels in multiple organs and whole blood of both mouse models...
May 8, 2017: Journal of Clinical Investigation
https://www.readbyqxmd.com/read/28473830/the-length-of-n-glycans-of-recombinant-h5n1-hemagglutinin-influences-the-oligomerization-and-immunogenicity-of-vaccine-antigen
#17
Agnieszka Katarzyna Macioła, Maria Anna Pietrzak, Piotr Kosson, Mariusz Czarnocki-Cieciura, Krzysztof Śmietanka, Zenon Minta, Edyta Kopera
Hemagglutinin glycoprotein (HA) is a principle influenza vaccine antigen. Recombinant HA-based vaccines become a potential alternative for traditional approach. Complexity and variation of HA N-glycosylation are considered as the important factors for the vaccine design. The number and location of glycan moieties in the HA molecule are also crucial. Therefore, we decided to study the effect of N-glycosylation pattern on the H5 antigen structure and its ability to induce immunological response. We also decided to change neither the number nor the position of the HA glycosylation sites but only the glycan length...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28471403/pathogenesis-and-inhibition-of-flaviviruses-from-a-carbohydrate-perspective
#18
REVIEW
So Young Kim, Bing Li, Robert J Linhardt
Flaviviruses are enveloped, positive single stranded ribonucleic acid (RNA) viruses with various routes of transmission. While the type and severity of symptoms caused by pathogenic flaviviruses vary from hemorrhagic fever to fetal abnormalities, their general mechanism of host cell entry is similar. All pathogenic flaviviruses, such as dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, and Zika virus, bind to glycosaminglycans (GAGs) through the putative GAG binding sites within their envelope proteins to gain access to the surface of host cells...
May 4, 2017: Pharmaceuticals
https://www.readbyqxmd.com/read/28467637/n-glycosylation-of-the-%C3%AE-2-adrenergic-receptor-regulates-receptor-function-by-modulating-dimerization
#19
Xiaona Li, Mang Zhou, Wei Huang, Huaiyu Yang
N-glycosylation is a common post-translational modification of G protein-coupled receptors (GPCRs). However, it remains unknown how N-glycosylation affects GPCR signaling. β2 adrenergic receptor (β2 AR) has three N-glycosylation sites: Asn6, Asn15 at the N-terminus and Asn187 at the second extracellular loop (ECL2). Here we show that deletion of the N-glycan did not affect receptor expression and ligand binding. Deletion of the N-glycan at the N-terminus rather than Asn187 showed decreased effects on isoproterenol-promoted G protein dependent signaling, β-arrestin2 recruitment and receptor internalization...
May 3, 2017: FEBS Journal
https://www.readbyqxmd.com/read/28463659/genetic-analysis-of-human-parainfluenza-virus-type-3-obtained-in-croatia-2011-2015
#20
Tanja Košutić-Gulija, Anamarija Slovic, Sunčanica Ljubin-Sternak, Gordana Mlinarić-Galinović, Dubravko Forčić
PURPOSE: This study investigated the HPIV3 circulating strains in Croatia and whether the other parts of HPIV3 genome (F gene and HN 582 nucleotides fragment) could be equally suitable for genetic and phylogenetic analysis. METHODOLOGY: Clinical materials were collected in period 2011-2015 from children suffering from respiratory illnesses. In positive HPIV3 samples viral genome was partially amplified and sequenced for HN and F genes. Obtained sequences were analysed by phylogenetic analysis and genetic characterization was performed...
April 2017: Journal of Medical Microbiology
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