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https://www.readbyqxmd.com/read/28453257/one-filter-one-sample-and-the-n-and-o-glyco-proteo-me-towards-a-system-to-study-disorders-of-protein-glycosylation
#1
Kirsty Skeene, Matthew Walker, Graham Clarke, Ed Bergstrom, Paul Genever, Daniel Ungar, Jane Thomas-Oates
A method has been developed for release/isolation of O-glycans from glycoproteins in whole cell lysates, for mass spectrometric analysis. Cells are lysed in SDS, which is then exchanged for urea and ammonium bicarbonate in a centrifugal filter, before treating with NH4OH to release O-glycans. Following centrifugation, O-glycans are recovered in the filtrate. Sonication achieves O-glycan release in 1 h. Combining the established protocol for filter-aided N-glycan separation, here optimised for enhanced PNGase F efficiency, with the developed O-glycan release method allows analysis of both N- and O-glycans from one sample, in the same filter unit, from 0...
April 28, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28449072/functions-of-galectins-as-self-non-self-recognition-and-effector-factors
#2
Gerardo R Vasta, Chiguang Feng, Nuria González-Montalbán, Justin Mancini, Lishi Yang, Kelsey Abernathy, Graeme Frost, Cheyenne Palm
Carbohydrate structures on the cell surface encode complex information that through specific recognition by carbohydrate-binding proteins (lectins), modulates interactions between cells, cells and the extracellular matrix, or mediates recognition of potential microbial pathogens. Galectins are a family of ß-galactoside-binding lectins, which are evolutionary conserved and have been identified in most organisms, from fungi to invertebrates and vertebrates, including mammals. Since their discovery in the 1970s, their biological roles, initially understood as limited to recognition of endogenous carbohydrate ligands in embryogenesis and development, have expanded in recent years by the discovery of their roles in tissue repair and regulation of immune homeostasis...
April 25, 2017: Pathogens and Disease
https://www.readbyqxmd.com/read/28448610/characterization-and-expression-analysis-of-galnts-in-developing-strongylocentrotus-purpuratus-embryos
#3
Amber L Famiglietti, Zheng Wei, Thomas M Beres, Adina L Milac, Duy T Tran, Divya Patel, Robert C Angerer, Lynne M Angerer, Lawrence A Tabak
Mucin-type O-glycosylation is a ubiquitous posttranslational modification in which N-Acetylgalactosamine (GalNAc) is added to the hydroxyl group of select serine or threonine residues of a protein by the family of UDP-GalNAc:Polypeptide N-Acetylgalactosaminyltransferases (GalNAc-Ts; EC 2.4.1.41). Previous studies demonstrate that O-glycosylation plays essential roles in protein function, cell-cell interactions, cell polarity and differentiation in developing mouse and Drosophila embryos. Although this type of protein modification is highly conserved among higher eukaryotes, little is known about this family of enzymes in echinoderms, basal deuterostome relatives of the chordates...
2017: PloS One
https://www.readbyqxmd.com/read/28446609/glycosylation-of-the-core-of-the-hiv-1-envelope-subunit-protein-gp120-is-not-required-for-native-trimer-formation-or-viral-infectivity
#4
Ujjwal Rathore, Piyali Saha, Sannula Kesavardhana, Aditya Arun Kumar, Rohini Datta, Sivasankar Devanarayanan, Raksha Das, John R Mascola, Raghavan Varadarajan
The gp120 subunit of HIV-1 envelope (Env) protein is heavily glycosylated at approximately 25 glycosylation sites, of which ~7-8 are located in the V1/V2 and V3 variable loops and the others in the remaining core gp120 region. Glycans partially shield Env from recognition by the host immune system and also are believed to be indispensable for proper folding of gp120 and for viral infectivity. Previous attempts to alter glycosylation sites in Env typically involved mutating the glycosylated asparagine residues to structurally similar glutamines or to alanines...
April 26, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28445724/quantification-of-the-impact-of-the-hiv-1-glycan-shield-on-antibody-elicitation
#5
Tongqing Zhou, Nicole A Doria-Rose, Cheng Cheng, Guillaume B E Stewart-Jones, Gwo-Yu Chuang, Michael Chambers, Aliaksandr Druz, Hui Geng, Krisha McKee, Young Do Kwon, Sijy O'Dell, Mallika Sastry, Stephen D Schmidt, Kai Xu, Lei Chen, Rita E Chen, Mark K Louder, Marie Pancera, Timothy G Wanninger, Baoshan Zhang, Anqi Zheng, S Katie Farney, Kathryn E Foulds, Ivelin S Georgiev, M Gordon Joyce, Thomas Lemmin, Sandeep Narpala, Reda Rawi, Cinque Soto, John-Paul Todd, Chen-Hsiang Shen, Yaroslav Tsybovsky, Yongping Yang, Peng Zhao, Barton F Haynes, Leonidas Stamatatos, Michael Tiemeyer, Lance Wells, Diana G Scorpio, Lawrence Shapiro, Adrian B McDermott, John R Mascola, Peter D Kwong
While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character)...
April 25, 2017: Cell Reports
https://www.readbyqxmd.com/read/28445718/the-hiv-1-glycan-shield-strategically-placed-kinks-in-the-armor-improve-antigen-design
#6
Christina Beatrice Karsten, Galit Alter
Dense glycosylation on the HIV-1 envelope glycoprotein hampers the induction of broadly neutralizing antibodies against HIV-1. Zhou et al. remove key glycans to unmask sites of vulnerability and enable the induction of neutralizing antibodies.
April 25, 2017: Cell Reports
https://www.readbyqxmd.com/read/28444931/study-of-structure-dependent-chromatographic-behavior-of-glycopeptides-using-reversed-phase-nanolc
#7
Petr Kozlik, Radoslav Goldman, Miloslav Sanda
Analysis of glycosylation is challenging due to micro- and macro-heterogeneity of the protein attachment. A combination of liquid chromatography with tandem mass spectrometry is one of the most powerful tools for glycopeptide analysis. In this work, we show the effect of various monosaccharide units on the retention time of glycopeptides. Retention behavior of several glycoforms of six peptides obtained from tryptic digest of haptoglobin, hemopexin and sex hormone binding globulin was studied on a reversed phase chromatographic column...
April 26, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28444921/biochemical-characterization-and-analyses-of-polysialic-acid-associated-carrier-proteins-and-genes-in-piglets-during-neonatal-development
#8
Bing Wang, Yue Chen, He Ren, Nai Zhang, Frederic Arthur Troy Ii
Polysialic acid plays a key role in cancer metastasis and neurodevelopment. Our aim was to determine the developmental gene expression profiles for the two polysialyltransferases ST 8Sia II and ST8Sia IV, NCAM, SynCAM1, Neuropilin-2 (NRP2) and their polysialylated cognate glycans in different regions of the piglet brain during postnatal development. Our findings show that: (1) The cellular levels of mRNA coding for ST8Sia II and ST8Sia IV, NCAM, SynCAM1, NRP2 and polySia are age-dependent and cell type specific during neonatal brain development; (2) There was a lack of correlation between abundance level of mRNA coding for ST8Sia II and ST8Sia IV and the abundance level of the post-translation expression of polySia in all nine brain regions; (3) Expression levels of polySia did not correlate with the levels of the carrier proteins, NCAM-140, SynCAM1 and NRP2 in nine brain regions; (4) The cellular abundance of ST8Sia II and ST8Sia IV in nine subregions of piglet brain is regulated at the level of translation/posttranslation, and not at the level of transcription...
April 26, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28444691/maldi-ms-profiling-of-serum-o-and-n-glycosylation-in-cog5-cdg
#9
Angelo Palmigiano, Rosaria Ornella Bua, Rita Barone, Daisy Rymen, Luc Régal, Nicolas Deconinck, Carlo Dionisi-Vici, Cheuk-Wing Fung, Domenico Garozzo, Jaak Jaeken, Luisa Sturiale
Congenital disorders of glycosylation (CDG) are due to defective glycosylation of glycoconjugates. COG-CDG are genetic diseases due to defects of the conserved oligomeric Golgi (COG) complex subunits 1-8 causing N- and O-glycan processing abnormalities. In COG-CDG, IEF separation of undersialylated glycoforms of serum transferrin and apolipoprotein C-III (apoC-III) allows to detect N- and O-glycosylation defects respectively. COG5-CDG (COG5 subunit deficiency) is a multisystem disease with dysmorphic features, intellectual disability of variable degree, seizures, acquired microcephaly, sensory defects and autistic behavior...
April 25, 2017: Journal of Mass Spectrometry: JMS
https://www.readbyqxmd.com/read/28443888/sugar-peptidic-bond-interactions-spectroscopic-characterization-of-a-model-system
#10
Ander Camiruaga, Imanol Usabiaga, Aran Insausti, Iker León, José A Fernández
Sugars are small carbohydrates which play numerous roles in living organisms such as storage of energy or as structural components. Modifications of specific sites within the glycan chain can modulate a carbohydrate's overall biological function as it happens with nucleic acids and proteins. Hence, identifying discrete carbohydrate modifications and understanding their biological effects is essential. A study of such processes requires of a deep knowledge of the interaction mechanism at the molecular level...
April 26, 2017: Physical Chemistry Chemical Physics: PCCP
https://www.readbyqxmd.com/read/28443077/molecular-dynamics-simulation-of-the-influenza-a-h3n2-hemagglutinin-trimer-reveals-the-structural-basis-for-adaptive-evolution-of-the-recent-epidemic-clade-3c-2a
#11
Masaru Yokoyama, Seiichiro Fujisaki, Masayuki Shirakura, Shinji Watanabe, Takato Odagiri, Kimito Ito, Hironori Sato
Influenza A(H3N2) has been a major cause of seasonal influenza in humans since 1968, and has evolved by antigenic drift under the constantly changing human herd immunity. Increasing evidence suggests that the antigenic change occasionally occurred concomitant with the alterations of the N-glycosylation site profile and hemagglutination activity of the virion surface protein hemagglutinin (HA). However, the structural basis of these changes remains largely unclear. To address this issue, we performed molecular dynamics simulations of the glycosylated HA trimers of the A(H3N2), which has a novel pattern of Asn-X-Ser/Thr sequons unique in the new A(H3N2) epidemic clade 3C...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28442549/neuraminidases-3-and-4-regulate-neuronal-function-by-catabolizing-brain-gangliosides
#12
Xuefang Pan, Camila De Britto Pará De Aragão, Juan P Velasco-Martin, David A Priestman, Harry Y Wu, Kohta Takahashi, Kazunori Yamaguchi, Luisella Sturiale, Domenico Garozzo, Frances M Platt, Nathalie Lamarche-Vane, Carlos R Morales, Taeko Miyagi, Alexey V Pshezhetsky
Gangliosides (sialylated glycolipids) play an essential role in the CNS by regulating recognition and signaling in neurons. Metabolic blocks in processing and catabolism of gangliosides result in the development of severe neurologic disorders, including gangliosidoses manifesting with neurodegeneration and neuroinflammation. We demonstrate that 2 mammalian enzymes, neuraminidases 3 and 4, play important roles in catabolic processing of brain gangliosides by cleaving terminal sialic acid residues in their glycan chains...
April 25, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28441734/chloroviruses-have-a-sweet-tooth
#13
REVIEW
James L Van Etten, Irina Agarkova, David D Dunigan, Michela Tonetti, Christina De Castro, Garry A Duncan
Chloroviruses are large double-stranded DNA (dsDNA) viruses that infect certain isolates of chlorella-like green algae. They contain up to approximately 400 protein-encoding genes and 16 transfer RNA (tRNA) genes. This review summarizes the unexpected finding that many of the chlorovirus genes encode proteins involved in manipulating carbohydrates. These include enzymes involved in making extracellular polysaccharides, such as hyaluronan and chitin, enzymes that make nucleotide sugars, such as GDP-L-fucose and GDP-D-rhamnose and enzymes involved in the synthesis of glycans attached to the virus major capsid proteins...
April 22, 2017: Viruses
https://www.readbyqxmd.com/read/28440416/fuca1-is-induced-by-wild-type-p53-and-expressed-at-different-levels-in-thyroid-cancers-depending-on-p53-status
#14
Nobuo Tsuchida, Masa-Aki Ikeda, Υoshizumu Ιshino, Michele Grieco, Giancarlo Vecchio
Fucose residues of cell surface glycans, which play important roles in growth, invasion and metastasis, are added by fucosyltransferases (FUTs) and removed by α-L-fucosidases (FUCAs). By the differential display method, we isolated a 3' non-coding region of α-L-fucosidase-1 (FUCA1) (a gene coding for the lysosomal fucosidase-1 enzyme) as a wild-type p53-inducible gene: 18S and 20S FUCA1 mRNA species were induced in Saos-2 cells transfected with a temperature-sensitive p53 mutant at the permissive temperature...
April 20, 2017: International Journal of Oncology
https://www.readbyqxmd.com/read/28438092/characterization-by-lectin-histochemistry-of-two-subpopulations-of-parietal-cells-in-the-rat-gastric-glands
#15
Laura Gómez-Santos, Edurne Alonso, Lucio Díaz-Flores, Juan F Madrid, Francisco J Sáez
Parietal cells undergo a differentiation process while they move from the isthmus toward the pits and the base region of the gastric gland. The aim of this work was to analyze the rat gastric glands by lectin histochemistry to show the glycans expressed by upper (young) and lower (old) parietal cells. We used lectins recognizing the most frequent sugar moieties in mammals. Each lectin was assayed alone and in combination with several deglycosylation pretreatments: (1) β-elimination, which removes O-linked oligosaccharides; (2) incubation with Peptide-N-glycosidase F, to remove N-linked glycans; (3) acid hydrolysis, which removes terminal sialic acid moieties; (4) methylation-saponification, to remove sulfate groups from sugar residues; and (5) glucose oxidase, a technique carried out with the lectin concanavalin A to convert glucose into gluconic acid...
May 2017: Journal of Histochemistry and Cytochemistry: Official Journal of the Histochemistry Society
https://www.readbyqxmd.com/read/28437766/cyanobacterial-lectins-characteristics-and-their-role-as-antiviral-agents
#16
REVIEW
Ram Sarup Singh, Amandeep Kaur Walia, Jasvirinder Singh Khattar, Davinder Pal Singh, John F Kennedy
Lectins are ubiquitous proteins/glycoproteins of non-immune origin that bind reversibly to carbohydrates in non-covalent and highly specific manner. These lectin-glycan interactions could be exploited for establishment of novel therapeutics, targeting the adherence stage of viruses and thus helpful in eliminating wide spread viral infections. Here the review focuses on the haemagglutination activity, carbohydrate specificity and characteristics of cyanobacterial lectins. Cyanobacterial lectins exhibiting high specificity towards mannose or complex glycans have potential role as anti-viral agents...
April 21, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28437635/three-cysteine-residues-of-slc52a1-a-receptor-for-the-porcine-endogenous-retrovirus-a-perv-a-play-a-critical-role-in-cell-surface-expression-and-infectivity
#17
Winston Colon-Moran, Takele Argaw, Carolyn A Wilson
Porcine endogenous retrovirus-A (PERV-A), a gammaretrovirus, infects human cells in vitro, thus raising the potential risk of cross-species transmission in xenotransplantation. Two members of the solute carrier family 52 (SLC52A1 and SLC52A2) are PERV-A receptors. Site-directed mutagenesis of the cDNA encoding SLC52A1 identified that only one of two putative glycosylation signals is occupied by glycans. In addition, we showed that glycosylation of SLC52A1 is not necessary for PERV-A receptor function. We also identified that at a minimum, three cysteine residues are sufficient for SLC52A1 cell surface expression...
April 20, 2017: Virology
https://www.readbyqxmd.com/read/28436597/glycan-analysis-by-ion-mobility-mass-spectrometry
#18
Johanna Hofmann, Kevin Pagel
Carbohydrates form one of the major classes of biological macromolecules in living organisms. To investigate their properties and function, an in-depth knowledge of their underlying structure is essential. However, the inherent structural complexity of glycans represents a major challenge. Carbohydrates are often branched and exhibit diverse regio- and stereochemistry. This in turn leads to a vast number of possible isomers, which are difficult to distinguish using established analytical tools. In the last decade, ion mobility-mass spectrometry, a technique that separates ions based on their mass, charge, size and shape, emerged as a powerful alternative for isomer distinction...
April 24, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28436457/fasciola-hepatica-glycoconjugates-immuneregulate-dendritic-cells-through-the-dendritic-cell-specific-intercellular-adhesion-molecule-3-grabbing-non-integrin-inducing-t-cell-anergy
#19
Ernesto Rodríguez, Hakan Kalay, Verónica Noya, Natalie Brossard, Cecilia Giacomini, Yvette van Kooyk, Juan J García-Vallejo, Teresa Freire
Dendritic cell-specific ICAM-3 grabbing non-integrin (DC-SIGN) expressed on a variety of DCs, is a C-type lectin receptor that recognizes glycans on a diverse range of pathogens, including parasites. The interaction of DC-SIGN with pathogens triggers specific signaling events that modulate DC-maturation and activity and regulate T-cell activation by DCs. In this work we evaluate whether F. hepatica glycans can immune modulate DCs via DC-SIGN. We demonstrate that DC-SIGN interacts with F. hepatica glycoconjugates through mannose and fucose residues...
April 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28436427/broad-and-potent-cross-clade-neutralizing-antibodies-with-multiple-specificities-in-the-plasma-of-hiv-1-subtype-c-infected-individuals
#20
Narayanaiah Cheedarla, K Lucia Precilla, Hemalatha Babu, K K Vidya Vijayan, Manickam Ashokkumar, Padmapriyadarsini Chandrasekaran, Nandagopal Kailasam, Jagadish Chandrabose Sundaramurthi, Soumya Swaminathan, Viswanath Buddolla, S Kalyanaraman Vaniambadi, V D Ramanathan, Luke Elizabeth Hanna
Broadly Cross clade Neutralizing (BCN) antibodies are recognized as potential therapeutic tools and leads for the design of a vaccine that can protect human beings against various clades of Human Immunodeficiency Virus (HIV). In the present study, we screened plasma of 88 HIV-1 infected ART naïve individuals for their neutralization potential using a standard panel of 18 pseudoviruses belonging to different subtypes and different levels of neutralization. We identified 12 samples with good breadth of neutralization (neutralized >90% of the viruses)...
April 24, 2017: Scientific Reports
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