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N-glycosylation

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https://www.readbyqxmd.com/read/28809486/assembling-glycan-charged-dolichol-phosphates-chemoenzymatic-synthesis-of-a-haloferax-volcanii-n-glycosylation-pathway-intermediate
#1
Yifat Elharar, Ananda R Podilapu, Ziqiang Guan, Suvarn S Kulkarni, Jerry Eichler
N-glycosylation, the covalent attachment of glycans to select protein target Asn residues, is a post-translational modification performed by all three domains of life. In the halophilic archaea Haloferax volcanii, where understanding of this universal protein-processing event is relatively well-advanced, genes encoding the components of the Agl (archaeal glycosylation) pathway responsible for the assembly and attachment of an N-linked pentasaccharide have been identified. As elsewhere, the N-linked glycan is assembled on phosphodolichol carriers before transfer to target Asn residues...
August 15, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28807751/a-novel-pmca3-mutation-in-an-ataxic-patient-with-hypomorphic-phosphomannomutase-2-pmm2-heterozygote-mutations-biochemical-characterization-of-the-pump-defect
#2
Mattia Vicario, Tito Calì, Domenico Cieri, Francesca Vallese, Raissa Bortolotto, Raffaele Lopreiato, Francesco Zonta, Marta Nardella, Alessia Micalizzi, Dirk J Lefeber, Enza Maria Valente, Enrico Bertini, Giuseppe Zanotti, Ginevra Zanni, Marisa Brini, Ernesto Carafoli
The neuron-restricted isoform 3 of the plasma membrane Ca(2+) ATPase plays a major role in the regulation of Ca(2+) homeostasis in the brain, where the precise control of Ca(2+) signaling is a necessity. Several function-affecting genetic mutations in the PMCA3 pump associated to X-linked congenital cerebellar ataxias have indeed been described. Interestingly, the presence of co-occurring mutations in additional genes suggest their synergistic action in generating the neurological phenotype as digenic modulators of the role of PMCA3 in the pathologies...
August 11, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28805451/genetic-characterisation-and-analysis-of-infectious-bronchitis-virus-isolated-from-brazilian-flocks-between-2010-and-2015
#3
Claudia Carranza, Claudete S Astolfi-Ferreira, Silvana H Santander Parra, Luis F N Nuñez, Zoltan Penzes, Jorge L Chacón, Luiz Sesti, Ruy D Chacón, Antonio J Piantino Ferreira
Infectious bronchitis virus (IBV) variants in Brazil were isolated during 2010-2015 for epidemiological and molecular analysis to characterise the different variants and perform a bioinformatic analysis to compare with sequences of variants collected over the previous 40 years. Of the 453 samples examined, 61.4% were positive for IBV and 75.9% of these were considered to have the BR-I genotype and were detected in birds of all ages distributed in all five Brazilian regions. The ratio of non-synonymous substitutions per non-synonymous site (dN) to synonymous substitutions per synonymous site (dS), i...
August 14, 2017: British Poultry Science
https://www.readbyqxmd.com/read/28803068/improved-therapeutic-efficacy-of-mammalian-expressed-recombinant-interferon-gamma-against-ovarian-cancer-cells
#4
Ali Razaghi, Carina Villacrés, Vincent Jung, Narges Mashkour, Michael Butler, Leigh Owens, Kirsten Heimann
Human interferon gamma (hIFNγ) affects tumour cells and modulates immune responses, showing promise as an anti-cancer biotherapeutic. This study investigated the effect of glycosylation and expression system of recombinant hIFNγ in ovarian carcinoma cell lines, PEO1 and SKOV3. The efficacy of E. coli- and mammalian-expressed hIFNγ (hIFNγ-CHO and HEK293, glycosylated/de-glycosylated) on cytostasis, cell death (MTT, and Guava-ViaCount(®) flow-cytometry) and apoptotic signalling (Western blot of Cdk2, histone H3, procaspase-3, FADD, cleaved PARP, and caspase-3) was examined...
August 9, 2017: Experimental Cell Research
https://www.readbyqxmd.com/read/28797655/n-glycan-profiles-in-h9n-2-avian-influenza-viruses-from-chicken-eggs-and-human-embryonic-lung-fibroblast-cells
#5
Wentian Chen, Yaogang Zhong, Rui Su, Huicai Qi, Weina Deng, Yu Sun, Tianran Ma, Xilong Wang, Hanjie Yu, Xiurong Wang, Zheng Li
N-glycosylation can affect the host specificity, virulence and infectivity of influenza A viruses (IAVs). In this study, the distribution and evolution of N-glycosylation sites in the hemagglutinin (HA) and neuraminidase (NA) of H9N2 virus were explored using phylogenetic analysis. Then, one strain of the H9N2 subtypes was proliferated in the embryonated chicken eggs (ECE) and human embryonic lung fibroblast cells (MRC-5) system. The proliferated viral N-glycan profiles were analyzed by a glycomic method that combined the lectin microarray and MALDI-TOF/TOF-MS...
August 7, 2017: Journal of Virological Methods
https://www.readbyqxmd.com/read/28794918/sensitive-and-structure-informative-n-glycosylation-analysis-by-maldi-ms-ionization-fragmentation-and-derivatization
#6
Takashi Nishikaze
Mass spectrometry (MS) has become an indispensable tool for analyzing post translational modifications of proteins, including N-glycosylated molecules. Because most glycosylation sites carry a multitude of glycans, referred to as "glycoforms," the purpose of an N-glycosylation analysis is glycoform profiling and glycosylation site mapping. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has unique characteristics that are suited for the sensitive analysis of N-glycosylated products...
2017: Mass Spectrometry
https://www.readbyqxmd.com/read/28780317/enzymatic-synthesis-of-ribo-and-2-deoxyribonucleosides-from-glycofuranosyl-phosphates-an-approach-to-facilitate-isotopic-labeling
#7
Wenhui Zhang, Toby Turney, Ivana Surjancev, Anthony S Serianni
Milligram quantities of α-D-ribofuranosyl 1-phosphate (sodium salt) (αR1P) were prepared by the phosphorolysis of inosine, catalyzed by purine nucleoside phosphorylase (PNPase). The αR1P was isolated by chromatography in >95% purity and characterized by (1)H and (13)C NMR spectroscopy. Aqueous solutions of αR1P were stable at pH 6.4 and 4 °C for several months. The isolated αR1P was N-glycosylated with different nitrogen bases (adenine, guanine and uracil) using PNPase or uridine phosphorylase (UPase) to give the corresponding ribonucleosides in high yield based on the glycosyl phosphate...
July 20, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/28777499/alg13-cdg-in-a-male-with-seizures-normal-cognitive-development-and-normal-transferrin-isoelectric-focusing
#8
Therese E Gadomski, Melody Bolton, Majid Alfadhel, Chris Dvorak, Olalekan A Ogunsakin, Stephen L Nelson, Eva Morava
ALG13-CDG has been recently discovered as a disorder of severe developmental, intellectual and speech disability, microcephaly, visual abnormalities, seizures, hepatomegaly, coagulation abnormalities, and abnormal serumtransferrin isoelectric focusing in serum. A male with seizures, delayed motor, and speech development, but normal cognition carried a hemizygous, predicted pathogenic ALG13 variant (p.E463G). N-glycosylation studies in plasma were normal. ICAM-1 expression was decreased in patient fibroblasts, supporting the variant's pathogenicity...
August 4, 2017: American Journal of Medical Genetics. Part A
https://www.readbyqxmd.com/read/28770921/insights-into-the-giardia-intestinalis-enolase-and-human-plasminogen-interaction
#9
R Aguayo-Ortiz, P Meza-Cervantez, R Castillo, A Hernández-Campos, L Dominguez, L Yépez-Mulia
Giardia intestinalis is an intestinal parasite that causes diarrhea in humans and animals worldwide. The enolase of G. intestinalis (GiENO) participates in its glycolysis pathway and is abundantly expressed in the parasite cytosol; however, its localization on the surface of trophozoites and cysts has been demonstrated. Enolases from bacteria and parasites can have different functions and are considered moonlighting proteins, for example, as a cell surface plasminogen receptor. In relation to GiENO, no studies have been performed about its possible participation as a plasminogen receptor...
August 3, 2017: Molecular BioSystems
https://www.readbyqxmd.com/read/28758349/appearance-of-us-like-porcine-epidemic-diarrhoea-virus-pedv-strains-before-us-outbreaks-and-genetic-heterogeneity-of-pedvs-collected-in-northern-vietnam-during-2012-2015
#10
N V Diep, M Sueyoshi, U Izzati, N Fuke, A P P Teh, N T Lan, R Yamaguchi
Porcine epidemic diarrhoea virus (PEDV) is the aetiologic agent of porcine epidemic diarrhoea (PED), a highly contagious enteric disease that is threatening the swine industry globally. Since PED was first reported in Southern Vietnam in 2009, the disease has spread throughout the country and caused substantial economic losses. To identify PEDVs responsible for the recent outbreaks, the full-length spike (S) gene of 25 field PEDV strains collected from seven northern provinces of Vietnam was sequenced and analysed...
July 30, 2017: Transboundary and Emerging Diseases
https://www.readbyqxmd.com/read/28755501/promoter-choice-impacts-the-efficiency-of-plant-glyco-engineering
#11
Somanath Kallolimath, Clemens Gruber, Herta Steinkellner, Alexandra Castilho
Glyco-modulation of therapeutic proteins produced in plants has shown great success. Plant-based expression platforms for tailored human-like N-glycosylation are based on the overexpression of foreign genes. However, drawbacks such as protein miss targeting, interference with endogenous glycosyltransferases or with plant development hamper the widespread use of the technology. Here we describe a technique that facilitates the generation of recombinant proteins with targeted N-glycosylation at high homogeneity...
July 29, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28753226/golgi-%C3%AE-1-4-fucosyltransferase-of-arabidopsis-thaliana-partially-localizes-at-the-nuclear-envelope
#12
Stephan Rips, Manuel Frank, Annegret Elting, Jan Niklas Offenborn, Antje von Schaewen
We analyzed plant-derived α1,4-Fucosyltransferase (FucTc) homologs by reporter fusions and focused on representatives of the Brassicaceae and Solanaceae. Arabidopsis thaliana AtFucTc-GFP or tomato LeFucTc-GFP restored Lewis-a formation in a fuctc mutant, confirming functionality in the trans-Golgi. AtFucTc-GFP partly accumulated at the nuclear envelope (NE) not observed for other homologs or truncated AtFucTc lacking the N-terminus or catalytic domain. Analysis of At/LeFucTc-GFP swap constructs with exchanged cytosolic, transmembrane and stalk (CTS), or only the CT regions, revealed that sorting information resides in the membrane anchor...
July 28, 2017: Traffic
https://www.readbyqxmd.com/read/28752594/serum-n-glycans-outperform-ca19-9-in-diagnosis-of-extrahepatic-cholangiocarcinoma
#13
Mengmeng Wang, Meng Fang, Jianhui Zhu, Huijuan Feng, Elisa Warner, Changhong Yi, Jun Ji, Xing Gu, Chunfang Gao
Extensive efforts have been devoted to improve the diagnosis of extrahepatic cholangiocarcinoma (ECCA) due to its silent clinical character and lack of effective diagnostic biomarkers. Specific alterations in N-glycosylation of glycoproteins are considered a key component in cancer progression, which can serve as a distinct molecular signature for cancer detection. This study aims to find potential serum N-glycan markers for ECCA. In total, 255 serum samples from patients with ECCA (n = 106), benign bile tract disease (BBD, n = 60) and healthy control (HC, n = 89) were recruited...
July 28, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28752167/a-novel-enzymatic-method-for-synthesis-of-glycopeptides-carrying-natural-eukaryotic-n-glycans
#14
Yangyang Xu, Zhigang Wu, Peiru Zhang, He Zhu, Hailiang Zhu, Qitao Song, Li Wang, Faxing Wang, Peng George Wang, Jiansong Cheng
A novel enzymatic approach was developed for facile production of glycopeptides carrying natural eukaryotic N-glycans. In this approach, peptides can be GlcNAcylated at one or two natural N-glycosylation sites via two-step enzymatic reactions catalyzed by an evolved N-glycosyltransferase (ApNGT(Q469A)) and a glucosamine N-acetyltransferase (GlmA), respectively. The resulting GlcNAc-peptides were further modified by an endo-β-N-acetylglucosaminidase M mutant (EndoM(N175Q)) to generate glycopeptides. In three steps of enzymatic catalysis, glycopeptides carrying complex-type N-glycans can be efficiently synthesized...
July 28, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28749929/chemoenzymatic-synthesis-of-glycoengineered-igg-antibodies-and-glycosite-specific-antibody-drug-conjugates
#15
Feng Tang, Lai-Xi Wang, Wei Huang
Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering...
August 2017: Nature Protocols
https://www.readbyqxmd.com/read/28746350/differential-effects-of-n-linked-glycosylation-of-vstm5-at-multiple-sites-on-surface-expression-and-filopodia-formation
#16
A-Ram Lee, Sulgi Kim, Kwang Woo Ko, Chul-Seung Park
V-set and transmembrane domain-containing protein 5 (Vstm5), a newly characterized small membrane glycoprotein, can induce membrane protrusions in various cells. Vstm5 can modulate both the position and complexity of central neurons by altering their membrane morphology and dynamics. In this study, we investigated the significance of glycosylation in the expression and function of Vstm5. Four N-linked glycosylation sites (Asn43, Asn87, Asn101, and Asn108) are predicted to be located in the extracellular N-terminus of mouse Vstm5...
2017: PloS One
https://www.readbyqxmd.com/read/28742884/reduced-expression-of-e-cadherin-and-increased-sialylation-level-in-clear-cell-renal-cell-carcinoma
#17
Małgorzata Borzym-Kluczyk, Iwona Radziejewska, Marzanna Cechowska-Pasko, Barbara Darewicz
Cancer cells are characterized by an aberrant increase in protein N-glycosylation and by disruption of E-cadherin-mediated adherens junctions. However, the relationship between alterations in N-glycosylation process and loss of E-cadherin adhesion in cancer remains unclear. The mechanisms of altered expression of adhesive glycoproteins in cancer cells have not been fully elucidated. Thus, the aim of this study was to examine the expression of E-cadherin and sialyl Lewis(a)/(x), NeuAcα2-3Gal, NeuAcα2-6Gal/GalNAc structures in the normal renal tissue and intermediate and cancerous tissues from patients with clear cell RCC...
July 24, 2017: Acta Biochimica Polonica
https://www.readbyqxmd.com/read/28738888/genetic-characterization-of-11-porcine-reproductive-and-respiratory-syndrome-virus-isolates-in-south-china-from-2014-to-2015
#18
Linyang Yu, Pandeng Zhao, Jianguo Dong, Yanling Liu, Leyi Zhang, Pengshuai Liang, Lei Wang, Changxu Song
BACKGROUND: Porcine reproductive and respiratory syndrome (PRRS) has leaded to an enormous loss per year to the swine industry, its etiology porcine reproductive and respiratory syndrome virus (PRRSV) is a highly mutated virus in pigs. To fully understand the genetic characteristics of PRRSV genome in South China, this study collected the lung samples infected with PRRSV in Guangdong and Hainan province from 2014 to 2015 and tried to isolate the PRRSV. Finally, the complete genomes of isolated strains were sequenced and analyzed...
July 24, 2017: Virology Journal
https://www.readbyqxmd.com/read/28737213/identification-of-methylated-gnti-dependent-n-glycans-in-botryococcus-brauni
#19
Stefan Schulze, Eugen Urzica, Maarten J M F Reijnders, Henri van de Geest, Sven Warris, Linda V Bakker, Christian Fufezan, Vitor A P Martins Dos Santos, Peter J Schaap, Sander A Peters, Michael Hippler
In contrast to mammals and vascular plants, microalgae show a high diversity in the N-glycan structures of complex N-glycoproteins. Although homologues for β1,2-N-acetylglucosaminyltransferase I (GnTI), a key enzyme in the formation of complex N-glycans, have been identified in several algal species, GnTI-dependent N-glycans have not been detected so far. We have performed an N-glycoproteomic analysis of the hydrocarbon oils accumulating green microalgae Botryococcus braunii. Thereby, the analysis of intact N-glycopeptides allowed the determination of N-glycan compositions...
September 2017: New Phytologist
https://www.readbyqxmd.com/read/28735024/n-linked-glycosylation-of-the-p24-family-protein-p24%C3%AE-5-modulates-retrograde-golgi-to-er-transport-of-k-hdel-ligands-in-arabidopsis
#20
Noelia Pastor-Cantizano, María Jesús García-Murria, Cesar Bernat-Silvestre, María Jesús Marcote, Ismael Mingarro, Fernando Aniento
The K/HDEL receptor ERD2 mediates the transport of soluble endoplasmic reticulum (ER)-resident proteins containing a C-terminal K/HDEL signal from the Golgi apparatus back to the ER via COPI (COat Protein I)-coated vesicles. Sorting of ERD2 within COPI vesicles is facilitated by p24 proteins. In Arabidopsis, p24δ5 has been shown to interact directly with ERD2 via its luminal GOLD (GOLgi Dynamics) domain and with COPI proteins via its cytoplasmic C-terminal tail at the acidic pH of the Golgi apparatus. Several members of the p24 family in mammals and yeast have been shown to be glycosylated, but whether Arabidopsis p24 proteins are glycosylated and the role of the sugar moiety in p24 function remain unclear...
August 7, 2017: Molecular Plant
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