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https://www.readbyqxmd.com/read/28436027/production-and-characterization-of-active-recombinant-human-factor-ii-with-consistent-sialylation
#1
Jeong H Lee, Jason Reier, Kelley M Heffner, Christopher Barton, David Spencer, Albert E Schmelzer, Raghavan Venkat
Coagulation factor II (prothrombin; FII) is the pre-proteolyzed precursor to thrombin in the coagulation cascade. It has ten sites of gamma-carboxylation, which are required for its bioactivity, and is N-glycosylated at three of four putative sites. Production of recombinant human FII (rhFII) using a platform fed-batch process designed for monoclonal antibody production resulted in low levels of gamma-carboxylation and sialylation. There have not been any prior reports of successful process development and clinical manufacture of rhFII with optimal, consistent gamma-carboxylation and sialylation...
April 24, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28431467/influence-of-glycosylation-on-interfacial-properties-of-recombinant-mucins-adsorption-surface-forces-and-friction
#2
Junxue An, Chunsheng Jin, Andra Dédinaité, Jan Holgersson, Niclas G Karlsson, Per Martin Claesson
Interfacial properties of two brush-with-anchor mucins, C-P55 and C-PSLex, have been investigated at the aqueous solution/poly(methylmethacrylate) (PMMA) interface. Both are recombinant mucin-type fusion proteins, produced by fusing the glycosylated mucin part of P-selectin glycoprotein ligand-1 (PSLG-1) to the Fc part of a mouse immunoglobulin in two different cells. They are mainly expressed as dimers upon production. Analysis of the O-glycans shows that the C-PSLex mucin has the longer and more branched side chains, but C-P55 has slightly higher sialic acid content...
April 21, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28431252/engineered-regulatory-systems-modulate-gene-expression-of-human-commensals-in-the-gut
#3
Bentley Lim, Michael Zimmermann, Natasha A Barry, Andrew L Goodman
The gut microbiota is implicated in numerous aspects of health and disease, but dissecting these connections is challenging because genetic tools for gut anaerobes are limited. Inducible promoters are particularly valuable tools because these platforms allow real-time analysis of the contribution of microbiome gene products to community assembly, host physiology, and disease. We developed a panel of tunable expression platforms for the prominent genus Bacteroides in which gene expression is controlled by a synthetic inducer...
April 20, 2017: Cell
https://www.readbyqxmd.com/read/28427293/beneficial-effects-of-tyrosol-on-altered-glycoprotein-components-in-streptozotocin-induced-diabetic-rats
#4
Ramasamy Chandramohan, Settu Saravanan, Leelavinothan Pari
CONTEXT: Olive oil is the major source of tyrosol which is a natural phenolic antioxidant. Olive oil constitutes a major component of the Mediterranean diet that is linked to a reduced incidence of chronic diseases. OBJECTIVE: This study evaluates the effects of tyrosol on altered glycoprotein components in streptozotocin-induced diabetic rats. MATERIALS AND METHODS: Diabetes mellitus was induced in male Wistar rats by streptozotocin (40 mg/kg body weight)...
December 2017: Pharmaceutical Biology
https://www.readbyqxmd.com/read/28426220/fluorescent-neuraminidase-assay-based-on-supramolecular-dye-capture-after-enzymatic-cleavage
#5
Wenqi Liu, César F A Gómez-Durán, Bradley D Smith
A conceptually new type of enzymatic cleavage assay is reported that utilizes in-situ supramolecular capture of the fluorescent product. A squaraine-derived substrate with large blocking groups at each end of its structure cannot be threaded by a tetralactam macrocycle until the blocking groups are removed by enzyme cleavage. A prototype design responds to viral neuraminidase, an indicator of influenza infection, and also measures susceptibility of the sample to neuraminidase inhibitor drugs. The substrate structure incorporates three key features; (a) a bis(4-amino-3-hydroxyphenyl)squaraine core with bright deep-red fluorescence and excellent photostability, (b) an N-methyl group at each end of the squaraine core that ensures fast macrocycle threading kinetics, and (c) sialic acid blocking groups that prevent macrocycle threading until they are removed by viral neuraminidase...
April 20, 2017: Journal of the American Chemical Society
https://www.readbyqxmd.com/read/28425925/evaluation-of-the-immunomodulatory-properties-of-streptococcus-suis-and-group-b-streptococcus-capsular-polysaccharides-on-the-humoral-response
#6
Cynthia Calzas, Morgan Taillardet, Insaf Salem Fourati, David Roy, Marcelo Gottschalk, Hugo Soudeyns, Thierry Defrance, Mariela Segura
Streptococcus suis and group B Streptococcus (GBS) are encapsulated streptococci causing septicemia and meningitis. Antibodies (Abs) against capsular polysaccharides (CPSs) have a crucial protective role, but the structure/composition of the CPS, including the presence of sialic acid, may interfere with the generation of anti-CPS Ab responses. We investigated the features of the CPS-specific Ab response directed against S. suis serotypes 2 and 14 and GBS serotypes III and V after infection or immunization with purified native or desialylated CPSs in mice...
April 20, 2017: Pathogens
https://www.readbyqxmd.com/read/28424685/analysis-of-linear-antibody-epitopes-on-factor-h-and-cfhr1-using-sera-of-patients-with-autoimmune-atypical-hemolytic-uremic-syndrome
#7
Eszter Trojnár, Mihály Józsi, Katalin Uray, Dorottya Csuka, Ágnes Szilágyi, Danko Milosevic, Vesna D Stojanović, Brankica Spasojević, Krisztina Rusai, Thomas Müller, Klaus Arbeiter, Kata Kelen, Attila J Szabó, György S Reusz, Satu Hyvärinen, T Sakari Jokiranta, Zoltán Prohászka
INTRODUCTION: In autoimmune atypical hemolytic uremic syndrome (aHUS), the complement regulator factor H (FH) is blocked by FH autoantibodies, while 90% of the patients carry a homozygous deletion of its homolog complement FH-related protein 1 (CFHR1). The functional consequence of FH-blockade is widely established; however, the molecular basis of autoantibody binding and the role of CFHR1 deficiency in disease pathogenesis are still unknown. We performed epitope mapping of FH to provide structural insight in the autoantibody recruitment on FH and potentially CFHR1...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28424265/effects-of-altered-sialic-acid-biosynthesis-on-n-linked-glycan-branching-and-cell-surface-interactions
#8
Nam D Pham, Poh-Choo Pang, Soumya Krishnamurthy, Amberlyn M Wands, Paola Grassi, Anne Dell, Stuart M Haslam, Jennifer J Kohler
GNE myopathy is a rare muscle disorder associated with aging and is related to sporadic inclusion body myositis (sIBM), the most common acquired muscle disease of aging. While the cause of sIBM is unknown, GNE myopathy is associated with mutations in UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). GNE harbors two enzymatic activities required for biosynthesis of sialic acid in mammalian cells. Mutations to both GNE domains are linked to GNE myopathy. However, correlation between mutation-associated reductions in sialic acid production and disease severity is imperfect...
April 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28423240/biochemical-cellular-physiological-and-pathological-consequences-of-human-loss-of-n-glycolylneuraminic-acid
#9
Jonathan Okerblom, Ajit Varki
About 2-3 million years ago, Alu-mediated deletion of a critical exon in the CMAH gene became fixed in the hominin lineage ancestral to humans, possibly through a stepwise process of selection by pathogen targeting of the CMAH product (the sialic acid Neu5Gc), followed by reproductive isolation via female anti-Neu5Gc antibodies. Loss of CMAH has occurred independently in some other lineages, but is functionally intact in Old World primates, including our closest relatives, the chimpanzees. While the biophysical and biochemical ramifications of losing tens of millions of Neu5Gc hydroxyl groups at most cell surfaces remains poorly understood, there are multi-scale effects functionally relevant to both sides of the host-pathogen interface...
April 19, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28420168/comparative-study-of-blood-based-biomarkers-%C3%AE-2-3-sialic-acid-psa-and-phi-for-high-risk-prostate-cancer-detection
#10
Montserrat Ferrer-Batallé, Esther Llop, Manel Ramírez, Rosa Núria Aleixandre, Marc Saez, Josep Comet, Rafael de Llorens, Rosa Peracaula
Prostate Specific Antigen (PSA) is the most commonly used serum marker for prostate cancer (PCa), although it is not specific and sensitive enough to allow the differential diagnosis of the more aggressive tumors. For that, new diagnostic methods are being developed, such as PCA-3, PSA isoforms that have resulted in the 4K score or the Prostate Health Index (PHI), and PSA glycoforms. In the present study, we have compared the PHI with our recently developed PSA glycoform assay, based on the determination of the α2,3-sialic acid percentage of serum PSA (% α2,3-SA), in a cohort of 79 patients, which include 50 PCa of different grades and 29 benign prostate hyperplasia (BPH) patients...
April 17, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28418182/cyclometalated-iridium-iii-bipyridine-phenylboronic-acid-complexes-as-luminescent-probes-for-sialic-acids-and-bioimaging-reagents
#11
Hua-Wei Liu, Wendell Ho-Tin Law, Lawrence Cho-Cheung Lee, Jonathan Chun-Wai Lau, Kenneth Kam-Wing Lo
Sialic acids play important roles in mammalian development, cell-cell attachment, and signaling. Since cancer cells utilize their overexpressed sialylated antigens to propagate metastases, the development of probes for sialic acids is of high importance. Herein, we report three luminescent cyclometalated iridium(III) bipyridine complexes bearing a phenylboronic acid (PBA) moiety. Spectrophotometric titrations revealed that the PBA complexes displayed higher binding affinity to the most common sialic acid N-acetylneuraminic acid (Neu5Ac) compared with simple sugars that are commonly found on glycoproteins...
April 18, 2017: Chemistry, An Asian Journal
https://www.readbyqxmd.com/read/28416510/erythrocyte-sialoglycoproteins-engage-siglec-9-on-neutrophils-to-suppress-activation
#12
Anel Lizcano, Ismael Secundino, Simon Döhrmann, Ross Corriden, Cristina Rohena, Sandra Diaz, Pradipta Ghosh, Lingquan Deng, Victor Nizet, Ajit Varki
Healthy blood neutrophils are functionally quiescent in the bloodstream, have a short lifespan, and exit the circulation to carry out innate immune functions, or undergo rapid apoptosis and macrophage-mediated clearance to mitigate host tissue damage. Limitation of unnecessary intravascular neutrophil activation is also important to prevent serious inflammatory pathologies. Since neutrophils become easily activated after purification, we carried out ex vivo comparisons with neutrophils maintained in whole blood...
April 17, 2017: Blood
https://www.readbyqxmd.com/read/28413949/the-intrinsic-relationship-between-structure-and-function-of-the-sialyltransferase-st8sia-family-members
#13
Ri-Bo Huang, D Cheng, Bo Lu, S M Liao, Frederic A Troy Ii, Guo-Ping Zhou
As a subset of glycosyltransferases, the family of sialyltransferases catalyze transfer of sialic acid (Sia) residues to terminal non-reducing positions on oligosaccharide chains of glycoproteins and glycolipids, utilizing CMP-Neu5Ac as the activated sugar nucleotide donor. In the four known sialyltransferase families (ST3Gal, ST6Gal, ST6GalNAc and ST8Sia), the ST8Sia family catalyzes synthesis of si,8-linked sialic/polysialic acid (polySia) chains according to their acceptor specificity. We have determined 3D structural models of the ST8Sia family members, designated ST8Sia I(1), II(2), IV(4), V(5), and VI(6) using the Phyre2 server...
April 14, 2017: Current Topics in Medicinal Chemistry
https://www.readbyqxmd.com/read/28411453/inhibition-of-influenza-virus-infection-by-multivalent-pentacyclic-triterpene-functionalized-per-o-methylated-cyclodextrin-conjugates
#14
Zhenyu Tian, Longlong Si, Kun Meng, Xiaoshu Zhou, Yongmin Zhang, Demin Zhou, Sulong Xiao
Multivalent ligands that exhibit high binding affinity to influenza hemagglutinin (HA) trimer can block the interaction of HA with its sialic acid receptor. In this study, a series of multivalent pentacyclic triterpene-functionalized per-O-methylated cyclodextrin (CD) derivatives were designed and synthesized using 1, 3-dipolar cycloaddition click reaction. A cell-based assay showed that three compounds (25, 28 and 31) exhibited strong inhibitory activity against influenza A/WSN/33 (H1N1) virus. Compound 28 showed the most potent anti-influenza activity with IC50 of 4...
April 2, 2017: European Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28409865/retention-of-glycopeptides-analyzed-using-hydrophilic-interaction-chromatography-is-influenced-by-charge-and-carbon-chain-length-of-ion-pairing-reagent-for-mobile-phase
#15
Kenichiro Furuki, Toshimasa Toyo'oka
Characterization of the glycans of glycoproteins is essential for the development and production of biologics. Numerous methods are available for analyzing the glycans of glycoproteins directly and labeled glycans. Nevertheless, glycopeptides are difficult to resolve because of their exceptional complexity and the microheterogeneity of glycans. These properties represent technical challenges to efforts to insure the accurate characterization of biopharmaceuticals to comply with regulatory requirements. Therefore, we investigated the retention behavior of peptides and glycopeptides in hydrophilic interaction chromatography (HILIC)-mode HPLC in the presence of ion-pairing reagents...
April 13, 2017: Biomedical Chromatography: BMC
https://www.readbyqxmd.com/read/28408901/serine-rich-repeat-adhesins-contribute-to-streptococcus-gordonii-induced-maturation-of-human-dendritic-cells
#16
Eun Byeol Ko, Sun Kyung Kim, Ho Seong Seo, Cheol-Heui Yun, Seung Hyun Han
Dendritic cells (DCs) play a pivotal role in the induction of immunity by recognition, capture, process, and presentation of antigens from infectious microbes. Streptococcus gordonii is able to cause life-threatening systemic diseases such as infective endocarditis. Serine-rich repeat (SRR) glycoproteins of S. gordonii are sialic acid-binding adhesins mediating the bacterial adherence to the host and the development of infective endocarditis. Thus, the SRR adhesins are potentially involved in the bacterial adherence to DCs and the maturation and activation of DCs required for the induction of immunity to S...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28400591/pathogenicity-and-transmission-of-a-swine-influenza-a-h6n6-virus
#17
Hailiang Sun, Bryan S Kaplan, Minhui Guan, Guihong Zhang, Jianqiang Ye, Li-Ping Long, Sherry Blackmon, Chun-Kai Yang, Meng-Jung Chiang, Hang Xie, Nan Zhao, Jim Cooley, David F Smith, Ming Liao, Carol Cardona, Lei Li, George Peng Wang, Richard Webby, Xiu-Feng Wan
Subtype H6 influenza A viruses (IAVs) are commonly detected in wild birds and domestic poultry and can infect humans. In 2010, a H6N6 virus emerged in southern China, and since then, it has caused sporadic infections among swine. We show that this virus binds to α2,6-linked and α2,3-linked sialic acids. Mutations at residues 222 (alanine to valine) and 228 (glycine to serine) of the virus hemagglutinin (HA) affected its receptor-binding properties. Experiments showed that the virus has limited transmissibility between ferrets through direct contact or through inhalation of infectious aerosolized droplets...
April 12, 2017: Emerging Microbes & Infections
https://www.readbyqxmd.com/read/28397880/glycoengineering-of-pertuzumab-and-its-impact-on-the-pharmacokinetic-pharmacodynamic-properties
#18
Cheng Luo, Song Chen, Na Xu, Chi Wang, Wen Bo Sai, Wei Zhao, Ying Chun Li, Xiao Jing Hu, Hong Tian, Xiang Dong Gao, Wen Bing Yao
Pertuzumab is an antihuman HER2 antibody developed for HER2 positive breast cancer. Glycosylation profiles are always the important issue for antibody based therapy. Previous findings have suggested the impact of glycosylation profiles on the function of antibodies, like pharmacodynamics, antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). However, the roles of fucose and sialic acid in the function of therapeutic antibodies still need further investigation, especially the role of sialic acid in nonfucosylated antibodies...
April 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28395125/probing-the-cmp-sialic-acid-donor-specificity-of-two-human-%C3%AE-d-galactoside-sialyltransferases-st3gal-i-and-st6gal-i-selectively-acting-on-o-and-n-glycosylproteins
#19
Maxence Noel, Pierre-André Gilormini, Virginie Cogez, Nao Yamakawa, Dorothée Vicogne, Cédric Lion, Christophe Biot, Yann Guerardel, Anne Harduin-Lepers
Sialylation of glycoproteins and glycolipids catalyzed by sialyltransferases are enzymatic reactions taking place in the Golgi of mammalian cells whereby sialic acid residues are added at the non-reducing ends of oligosaccharides. Because sialylated glycans play critical roles in a number of human physio-pathological processes, the past two decades have witnessed the development of modified sialic acid derivatives for a better understanding of sialic acid biology and development of new therapeutic targets. However, nothing is known about how individual mammalian sialyltransferases tolerate and behave towards these unnatural CMP-sialic acid donors...
April 10, 2017: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/28393524/changes-in-caprine-milk-oligosaccharides-at-different-lactation-stages-analyzed-by-high-performance-liquid-chromatography-coupled-to-mass-spectrometry
#20
Andrea Martín-Ortiz, Daniela Barile, Jaime Salcedo, F Javier Moreno, Alfonso Clemente, Ana I Ruiz-Matute, María L Sanz
Changes of the abundance of caprine milk oligosaccharides (CMO) at different lactation stages have been evaluated by hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-Q MS) and nanoflow liquid chromatography-quadrupole-time-of-flight mass spectrometry (nano-LC-Chip-QTOF MS). Eight major oligosaccharides (OS) were quantified at different lactation stages by HILIC-Q MS, while the use of nano-LC-Chip-QToF MS allowed expanding the study to forty-nine different OS by monitoring neutral non- and fucosylated species, as well as acidic species containing not only N-acetyl-neuraminic acid or N-glycolyl-neuraminic acid residues but also the combination of both sialic acids...
April 20, 2017: Journal of Agricultural and Food Chemistry
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