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N-glycosylation

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https://www.readbyqxmd.com/read/28342150/expression-and-characterization-of-human-%C3%AE-1-4-galactosyltransferase-1-%C3%AE-4galt1-using-silkworm-baculovirus-expression-system
#1
Daisuke Morokuma, Jian Xu, Masato Hino, Hiroaki Mon, Jasmeen S Merzaban, Masateru Takahashi, Takahiro Kusakabe, Jae Man Lee
Baculovirus expression vector system (BEVS) is widely known as a mass-production tool to produce functional recombinant glycoproteins except that it may not be always suitable for medical practice due to the differences in the structure of N-linked glycans between insects and mammalian. Currently, various approaches have been reported to alter N-linked glycan structures of glycoproteins derived from insects into terminally sialylated complex-type N-glycans. In the light of those studies, we also proposed in vitro maturation of N-glycan with mass-produced and purified glycosyltransferases by silkworm-BEVS...
March 24, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28338695/unprecedented-highly-efficient-capture-of-glycopeptides-by-fe3o4-mg-mof-74-core-shell-nanoparticles
#2
Jie Li, Jiaxi Wang, Yun Ling, Zhenxia Chen, Mingxia Gao, Xiangmin Zhang, Yaming Zhou
Magnetic Fe3O4 nanospheres coated with Mg-MOF-74 have been facilely prepared via epitaxial growth. Owing to the intrinsic hydrophilic pore surface and size-exclusive properties of Mg-MOF-74, the Fe3O4@Mg-MOF-74 core-shell nanoparticles show effective and selective enrichment of 441 N-glycosylation sites of 418 glycopeptides from 125 glycoproteins in 1 μL of human serum.
March 24, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28336960/co-expression-of-bira-with-biotin-bait-achieves-in-vivo-biotinylation-of-overexpressed-stable-n-glycosylated-srage-in-transgenic-silkworms
#3
Miyuki Kumano-Kuramochi, Ken-Ichiro Tatematsu, Mayumi Ohnishi-Kameyama, Mari Maeda-Yamamoto, Toshiro Kobori, Hideki Sezutsu, Sachiko Machida
Here, we demonstrated the expression of the N-glycosylated extracellular ligand binding domain of receptor for advanced glycation end products (sRAGE) in middle silk glands (MSGs) of transgenic silkworms using the GAL4/UAS system. Over 1 mg of sRAGE was obtained from one transgenic silkworm. sRAGE purified from the silkworm exhibited good stability and maintained specific ligand-binding ability. In addition, N-glycan analysis of sRAGE revealed that N-glucan completely lacked potentially allergenic fucose...
March 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28336547/glycans-in-the-intestinal-peptide-transporter-pept1-contribute-to-function-and-protect-from-proteolysis
#4
Tamara Stelzl, Kerstin Elisabeth Geillinger-Kästle, Jürgen Stolz, Hannelore Daniel
Despite the fact that many membrane proteins carry extracellular glycans, little is known about whether the glycan chains also affect protein function. We recently demonstrated that the proton-coupled oligopeptide transporter 1 (PEPT1) in the intestine is glycosylated at six asparagine residues (N50, N406, N439, N510, N515, N532). Mutagenesis-induced disruption of the individual N-glycosylation site N50, which is highly conserved among mammals, was detected to significantly enhance the PEPT1 mediated inward transport of peptides...
March 23, 2017: American Journal of Physiology. Gastrointestinal and Liver Physiology
https://www.readbyqxmd.com/read/28334832/the-n-glycosylation-of-immunoglobulin-g-as-a-novel-biomarker-of-parkinson-s-disease
#5
Alyce C Russell, Mirna Šimurina, Monique T Garcia, Mislav Novokmet, Youxin Wang, Igor Rudan, Harry Campbell, Gordan Lauc, Meghan G Thomas, Wei Wang
The use of the emerging "omics" technologies for large scale population screening is promising in terms of predictive, preventive and personalized medicine. For Parkinson's disease, it is essential that an accurate diagnosis is obtained and disease progression can be monitored. Immunoglobulin G (IgG) has the ability to exert both anti-inflammatory and pro-inflammatory effects, and the N-glycosylation of the fragment crystallizable portion of IgG is involved in this process. This study aimed to determine whether the IgG glycome could be a candidate biomarker for Parkinson's disease...
March 17, 2017: Glycobiology
https://www.readbyqxmd.com/read/28332009/n-glycosylation-of-an-igg-antibody-secreted-by-nicotiana-tabacum-by-2-cells-can-be-modulated-through-co-expression-of-human-%C3%AE-1-4-galactosyltransferase
#6
Catherine Navarre, Nicolas Smargiasso, Laurent Duvivier, Joseph Nader, Johann Far, Edwin De Pauw, Marc Boutry
Nicotiana tabacum BY-2 suspension cells have several advantages that make them suitable for the production of full-size monoclonal antibodies which can be purified directly from the culture medium. Carbohydrate characterization of an antibody (Lo-BM2) expressed in N. tabacum BY-2 cells showed that the purified Lo-BM2 displays N-glycan homogeneity with a high proportion (>70%) of the complex GnGnXF glycoform. The stable co-expression of a human β-1,4-galactosyltransferase targeted to different Golgi sub-compartments altered Lo-BM2N-glycosylation and resulted in the production of an antibody that exhibited either hybrid structures containing a low abundance of the plant epitopes (α-1,3-fucose and β-1,2-xylose), or a large amount of galactose-extended N-glycan structures...
March 22, 2017: Transgenic Research
https://www.readbyqxmd.com/read/28331098/recombinant-modified-vaccinia-virus-ankara-generating-ebola-virus-like-particles
#7
Marc Schweneker, Andrea S Laimbacher, Gert Zimmer, Susanne Wagner, Elisabeth M Schraner, Michael Wolferstätter, Marieken Klingenberg, Ulrike Dirmeier, Robin Steigerwald, Henning Lauterbach, Hubertus Hochrein, Paul Chaplin, Mark Suter, Jürgen Hausmann
There are currently no approved therapeutics or vaccines to treat or protect against the severe hemorrhagic fever and death caused by Ebola virus (EBOV). Ebola virus-like particles (EBOV-VLPs) consisting of the matrix protein VP40, the glycoprotein (GP) and the nucleoprotein (NP) are highly immunogenic and protective in non-human primates against Ebola virus disease (EVD). We have constructed a modified vaccinia virus Ankara-Bavarian Nordic(®) (MVA-BN(®)) recombinant co-expressing VP40 and glycoprotein (GP) of EBOV Mayinga and the nucleoprotein (NP) of Taï Forest virus (TAFV) (MVA-BN-EBOV-VLP) to launch non-infectious EBOV-VLPs as a second vaccine modality in the MVA-BN-EBOV-VLP-vaccinated organism...
March 22, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28330764/lipid-sugar-carriers-at-the-extremes-the-phosphodolichols-archaea-use-in-n-glycosylation
#8
REVIEW
Jerry Eichler, Ziqiang Guan
N-glycosylation, a post-translational modification whereby glycans are covalently linked to select Asn residues of target proteins, occurs in all three domains of life. Across evolution, the N-linked glycans are initially assembled on phosphorylated cytoplasmically-oriented polyisoprenoids, with polyprenol (mainly C55 undecaprenol) fulfilling this role in Bacteria and dolichol assuming this function in Eukarya and Archaea. The eukaryal and archaeal versions of dolichol can, however, be distinguished on the basis of their length, degree of saturation and by other traits...
March 19, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28328971/mapping-the-n-linked-glycosites-of-rice-oryza-sativa-l-germinating-embryos
#9
Jiezheng Ying, Juan Zhao, Yuxuan Hou, Yifeng Wang, Jiehua Qiu, Zhiyong Li, Xiaohong Tong, Zhaomei Shi, Jun Zhu, Jian Zhang
Germination is a key event in the angiosperm life cycle. N-glycosylation of proteins is one of the most common post-translational modifications, and has been recognized to be an important regulator of the proteome of the germinating embryo. Here, we report the first N-linked glycosites mapping of rice embryos during germination by using a hydrophilic interaction chromatography (HILIC) glycopeptides enrichment strategy associated with high accuracy mass spectrometry identification. A total of 242 glycosites from 191 unique proteins was discovered...
2017: PloS One
https://www.readbyqxmd.com/read/28327546/missense-mutations-near-the-n-glycosylation-site-of-the-a2-domain-lead-to-various-intracellular-trafficking-defects-in-coagulation-factor-viii
#10
Wei Wei, Chunlei Zheng, Min Zhu, Xiaofan Zhu, Renchi Yang, Saurav Misra, Bin Zhang
Missense mutation is the most common mutation type in hemophilia. However, the majority of missense mutations remain uncharacterized. Here we characterize how hemophilia mutations near the unused N-glycosylation site of the A2 domain (N582) of FVIII affect protein conformation and intracellular trafficking. N582 is located in the middle of a short 310-helical turn (D580-S584), in which most amino acids have multiple hemophilia mutations. All 14 missense mutations found in this 310-helix reduced secretion levels of the A2 domain and full-length FVIII...
March 22, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28323990/galactose-supplementation-in-tmem165-cdg-patients-rescues-the-glycosylation-defects
#11
Willy Morelle, Sven Potelle, Peter Witters, Sunnie Wong, Leslie Climer, Vladimir Lupashin, Gert Matthijs, Therese Gadomski, Jaak Jaeken, David Cassiman, Eva Morava, François Foulquier
Context: TMEM165 deficiency is a severe multisystem disease that manifests with metabolic, endocrine and skeletal involvement. It leads to one type of congenital disorders of glycosylation (CDG), a rapidly growing group of inherited diseases where the glycosylation process is altered. Patients have decreased galactosylation by serum glycan analysis. There are over one hundred CDGs but only specific types are treatable. Objective: Galactose has been shown to be beneficial in other CDG types with abnormal galactosylation...
January 9, 2017: Journal of Clinical Endocrinology and Metabolism
https://www.readbyqxmd.com/read/28322444/identification-and-characterization-of-site-specific-n-glycosylation-in-the-potassium-channel-kv3-1b
#12
Paul Christian Vicente, Jin Young Kim, Jeong-Ju Ha, Min-Young Song, Hyun-Kyung Lee, Dong-Hyun Kim, Jin-Sung Choi, Kang-Sik Park
The potassium ion channel Kv3.1b is a member of a family of voltage-gated ion channels that are glycosylated in their mature form. In the present study, we demonstrate the impact of N-glycosylation at specific asparagine residues on the trafficking of the Kv3.1b protein. Large quantities of asparagine 229 (N229)-glycosylated Kv3.1b reached the plasma membrane, whereas N220-glycosylated and unglycosylated Kv3.1b were mainly retained in the endoplasmic reticulum (ER). These ER-retained Kv3.1b proteins were susceptible to degradation when co-expressed with calnexin, whereas Kv3...
March 21, 2017: Journal of Cellular Physiology
https://www.readbyqxmd.com/read/28321273/characterization-of-a-new-monoclonal-anti-glypican-3-antibody-specific-to-the-hepatocellular-carcinoma-cell-line-hepg2
#13
Preeyanat Vongchan, Robert J Linhardt
AIM: To characterize the antigen on HepG2 cell that is specifically recognized by a new monoclonal antibody raised against human liver heparan sulfate proteoglycan (HSPG), clone 1E4-1D9. METHODS: The antigen recognized by mAb 1E4-1D9 was immunoprecipitated and its amino acid sequence was analyzed LC/MS. The transmembrane domain, number of cysteine residues, and glycosylation sites were predicted from these entire sequences. Data from amino acid analysis was aligned with glypican-3 (https://www...
March 8, 2017: World Journal of Hepatology
https://www.readbyqxmd.com/read/28318221/crystal-structure-of-a-homogeneous-igg-fc-glycoform-with-the-n-glycan-designed-to-maximize-the-antibody-dependent-cellular-cytotoxicity
#14
Chia-Lin Chen, Jen-Chi Hsu, Chin-Wei Lin, Chia-Hung Wang, Ming-Hung Tsai, Chung-Yi Wu, Chi-Huey Wong, Che Ma
N-glycosylation on IgG modulates Fc conformation and effector functions. An IgG-Fc contains a human sialo-complex type (hSCT) glycan of biantennary structure with two α2,6-sialylations and without core-fucosylation is an optimized glycoform developed to enhance the antibody dependent cellular cytotoxicity (ADCC). hSCT modification not only enhances the binding affinity to Fc receptors in the presence of antigen, but also in some cases provides gain-of-function effector activity. We used enzymatic glyco-engineering to prepare an IgG-Fc with homogeneous hSCT attached to each CH2 domain, and solved its crystal structure...
March 20, 2017: ACS Chemical Biology
https://www.readbyqxmd.com/read/28315854/mutation-of-n-linked-glycosylation-in-epcam-affected-cell-adhesion-in-breast-cancer-cells
#15
Xue Liu, Jiujiao Gao, Yan Sun, Dandan Zhang, Tingjiao Liu, Qiu Yan, Xuesong Yang
EpCAM expression is elevated in breast cancer tissue, and correlates with the cancer metastasis and cell adhesion. Although EpCAM glycosylation is supposed to be associated with its function, the contribution of N-glycosylation to its function remains unclear. Here we analyzed cell adhesion ability of EpCAM in breast cancer cells. The results showed that EpCAM expression was associated with cell adhesion and N-glycosylation mutation of EpCAM decreased adhesion capacity. N-glycosylation mutation of EpCAM was correlated with lower levels of integrin β1 and fibronectin...
March 18, 2017: Biological Chemistry
https://www.readbyqxmd.com/read/28303575/isoform-localization-of-dectin-1-regulates-the-signaling-quality-of-anti-fungal-immunity
#16
Mike Fischer, Jörg P Müller, Bärbel Spies-Weisshart, Christine Gräfe, Oliver Kurzai, Kerstin Hünniger, Andreas Hochhaus, Sebastian Scholl, Ulf Schnetzke
Dectin-1 is recognized as a major receptor for fungal ß-glucans and contributes to anti-fungal immunity. Human monocyte populations express Dectin-1 isoforms A and B, which differ by the presence of a stalk region and its N-linked glycosylation site. Here, we analyzed the expression of both isoforms in human monocyte-derived cells. The cellular localization on cell lines stably expressing either Dectin-1 isoform A or B was studied by flow cytometry and confocal laser scanning microscopy. Intracellular protein signaling and cytokine production was analyzed by immunoblotting and cytometric bead arrays, respectively...
March 16, 2017: European Journal of Immunology
https://www.readbyqxmd.com/read/28300864/whole-exome-sequencing-in-the-framingham-heart-study-identifies-rare-variation-in-hyal2-that-influences-platelet-aggregation
#17
John D Eicher, Ming-Huei Chen, Achilleas N Pitsillides, Honghuang Lin, Narayanan Veeraraghavan, Jennifer A Brody, Ginger A Metcalf, Donna M Muzny, Richard A Gibbs, Diane M Becker, Lewis C Becker, Nauder Faraday, Rasika A Mathias, Lisa R Yanek, Eric Boerwinkle, L Adrienne Cupples, Andrew D Johnson
Inhibition of platelet reactivity is a common therapeutic strategy in secondary prevention of cardiovascular disease. Genetic and environmental factors influence inter-individual variation in platelet reactivity. Identifying genes that contribute to platelet reactivity can reveal new biological mechanisms and possible therapeutic targets. Here, we examined rare coding variation to identify genes associated with platelet reactivity in a population-based cohort. To do so, we performed whole exome sequencing in the Framingham Heart Study and conducted single variant and gene-based association tests against platelet reactivity to collagen, adenosine diphosphate (ADP), and epinephrine agonists in up to 1,211 individuals...
March 16, 2017: Thrombosis and Haemostasis
https://www.readbyqxmd.com/read/28299773/trafficking-and-other-regulatory-mechanisms-for-organic-anion-transporting-polypeptides-oatps-and-organic-anion-transporters-oats-that-modulate-cellular-drug-and-xenobiotic-influx-and-that-are-dysregulated-in-disease
#18
REVIEW
Michael Murray, Fanfan Zhou
Organic anion transporters (OATs) and organic anion-transporting polypeptides (OATPs), encoded by a number of SLC22A and SLCO genes, mediate the absorption and distribution of drugs and other xenobiotics. The regulation of OATs and OATPs is complex, comprising both transcriptional and post-translational mechanisms. Plasma membrane expression is required for cellular substrate influx by OATs/OATPs. Thus, interest in post-translational regulatory processes, including membrane targeting, endocytosis, recycling and degradation of transporter proteins, is increasing because these are critical for plasma membrane expression...
March 15, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28299519/insights-into-the-effects-of-glycosylation-and-the-monosaccharide-binding-activity-of-the-plant-lectin-cratabl
#19
Laercio Pol-Fachin
CrataBL is a glycoprotein isolated from Crataeva tapia bark, containing two N-glycosylation sites. It has been identified to present lectin activity with some specificity for binding glucose over galactose. However, to date, no information on the effects of glycosylation or CrataBL monosaccharide-binding sites and monosaccharide specificity has been obtained. Thus, molecular docking and molecular dynamics simulations were employed to characterize the glycosylated CrataBL conformation and dynamics in aqueous solutions, as well as the molecular basis for its binding specificity...
March 15, 2017: Glycoconjugate Journal
https://www.readbyqxmd.com/read/28298204/immunoglobulin-t-from-sea-bass-dicentrarchus-labrax-l-molecular-characterization-tissue-localization-and-expression-after-nodavirus-infection
#20
Francesco Buonocore, Valentina Stocchi, Noelia Nunez-Ortiz, Elisa Randelli, Marco Gerdol, Alberto Pallavicini, Angelo Facchiano, Chiara Bernini, Laura Guerra, Giuseppe Scapigliati, Simona Picchietti
BACKGROUND: Immunoglobulins (Igs) are fundamental components of the adaptive immune system of vertebrates, with the IgT/IgZ isotype specific of Teleosts. In this paper we describe the identification of an IgT heavy chain from the European sea bass (Dicentrarchus labrax L.), its molecular characterization and tissue mRNA localization by in situ hybridization. RESULTS: Sea bass IgT consists of 552 aa (Accession Number KM410929) and it contains a putative 19 amino acids long signal peptide and one potential N-glycosylation site...
March 15, 2017: BMC Molecular Biology
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