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N-glycosylation

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https://www.readbyqxmd.com/read/28932884/facile-and-easily-popularized-synthesis-of-l-cysteine-functionalized-magnetic-nanoparticles-based-on-one-step-functionalization-for-highly-efficient-enrichment-of-glycopeptides
#1
Xiaoyan Feng, Chunhui Deng, Mingxia Gao, Xiangmin Zhang
Protein glycosylation is one of the most important post-translational modifications. Also, efficient enrichment and separation of glycopeptides from complex samples are crucial for the thorough analysis of glycosylation. Developing novel hydrophilic materials with facile and easily popularized synthesis is an urgent need in large-scale glycoproteomics research. Herein, for the first time, a one-step functionalization strategy based on metal-organic coordination was proposed and Fe3O4 nanoparticles were directly functionalized with zwitterionic hydrophilic L-cysteine (L-Cys), greatly simplifying the synthetic procedure...
September 20, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28932688/alg9-cdg-new-clinical-case-and-review-of-the-literature
#2
Kellie Davis, Duncan Webster, Chris Smith, Sheryl Jackson, David Sinasac, Lorne Seargeant, Xing-Chang Wei, Patrick Ferreira, Julian Midgley, Yolanda Foster, Xueli Li, Miao He, Walla Al-Hertani
Congenital disorders of glycosylation (CDG) are a group of metabolic diseases resulting from defects in glycan synthesis or processing. The number of subgroups and their phenotypic spectrums continue to expand with most related to deficiencies of N-glycosylation. ALG9-CDG (previously CDG-IL) is the result of a mutation in ALG9. This gene encodes the enzyme alpha-1,2-mannosyltransferase. To date, a total of 10 patients from 6 different families have been reported with one of four ALG9 mutations. Seven of these patients had a similar phenotype with failure to thrive, dysmorphic features, seizures, hepatic and/or renal cysts; the other three patients died in utero from a lethal skeletal dysplasia...
December 2017: Molecular Genetics and Metabolism Reports
https://www.readbyqxmd.com/read/28931878/a-method-to-identify-trace-sulfated-igg-n-glycans-as-biomarkers-for-rheumatoid-arthritis
#3
Jing-Rong Wang, Wei-Na Gao, Rudolf Grimm, Shibo Jiang, Yong Liang, Hua Ye, Zhan-Guo Li, Lee-Fong Yau, Hao Huang, Ju Liu, Min Jiang, Qiong Meng, Tian-Tian Tong, Hai-Hui Huang, Stephanie Lee, Xing Zeng, Liang Liu, Zhi-Hong Jiang
N-linked glycans on immunoglobulin G (IgG) have been associated with pathogenesis of diseases and the therapeutic functions of antibody-based drugs; however, low-abundance species are difficult to detect. Here we show a glycomic approach to detect these species on human IgGs using a specialized microfluidic chip. We discover 20 sulfated and 4 acetylated N-glycans on IgGs. Using multiple reaction monitoring method, we precisely quantify these previously undetected low-abundance, trace and even ultra-trace N-glycans...
September 20, 2017: Nature Communications
https://www.readbyqxmd.com/read/28929175/tusc3-functional-duality-of-a-cancer-gene
#4
REVIEW
Kateřina Vašíčková, Peter Horak, Petr Vaňhara
Two decades ago, following a systematic screening of LOH regions on chromosome 8p22, TUSC3 has been identified as a candidate tumor suppressor gene in ovarian, prostate and pancreatic cancers. Since then, a growing body of evidence documented its clinical importance in various other types of cancers, and first initial insights into its molecular function and phenotypic effects have been gained, though the precise role of TUSC3 in different cancers remains unclear. As a part of the oligosaccharyltransferase complex, TUSC3 localizes to the endoplasmic reticulum and functions in final steps of N-glycosylation of proteins, while its loss evokes the unfolded protein response...
September 19, 2017: Cellular and Molecular Life Sciences: CMLS
https://www.readbyqxmd.com/read/28929132/interplay-between-protein-glycosylation-pathways-in-alzheimer-s-disease
#5
Moran Frenkel-Pinter, Merav Daniel Shmueli, Chen Raz, Michaela Yanku, Shai Zilberzwige, Ehud Gazit, Daniel Segal
Deviations from the normal nucleoplasmic protein O-GlcNAcylation, as well as from normal protein sialylation and N-glycosylation in the secretory pathway, have been reported in Alzheimer's disease (AD). However, the interplay between the cytoplasmic protein O-GlcNAcylation and the secretory N-/O-glycosylation in AD has not been described. We present a comprehensive analysis of the N-, O-, and O-GlcNAc-glycomes in AD-affected brain regions as well as in AD patient serum. We detected marked differences in levels of glycan involved in both protein O-GlcNAcylation and N-/O-glycosylation between patients and healthy individuals and revealed brain region-specific glycosylation-related pathology in patients...
September 2017: Science Advances
https://www.readbyqxmd.com/read/28928222/single-subunit-oligosaccharyltransferases-of-trypanosoma-brucei-display-different-and-predictable-peptide-acceptor-specificities
#6
Anders A J Jinnelov, Liaqat Ali, Michele Tinti, Michael A J Ferguson
Trypanosoma brucei causes African trypanosomiasis and contains three full-length oligosaccharyltransferase (OST) genes; two of which, TbSTT3A and TbSTT3B, are expressed in the bloodstream form of the parasite. These OSTs have different peptide acceptor and lipid-linked oligosaccharide donor specificities and trypanosomes do not follow many of the canonical rules developed for other eukaryotic N-glycosylation pathways, raising questions as to the basic architecture and detailed function of trypanosome OSTs. Here, we show by blue-native gel electrophoresis and stable isotope labelling in cell culture proteomics that the TbSTT3A and TbSTT3B proteins associate with each other in large complexes that contain no other detectable protein subunits...
September 19, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28928140/selenoprotein-t-is-a-novel-ost-subunit-that-regulates-upr-signaling-and-hormone-secretion
#7
Abdallah Hamieh, Dorthe Cartier, Houssni Abid, André Calas, Carole Burel, Christine Bucharles, Cedric Jehan, Luca Grumolato, Marc Landry, Patrice Lerouge, Youssef Anouar, Isabelle Lihrmann
Selenoprotein T (SelT) is a recently characterized thioredoxin-like protein whose expression is very high during development, but is confined to endocrine tissues in adulthood where its function is unknown. We report here that SelT is required for adaptation to the stressful conditions of high hormone level production in endocrine cells. Using immunofluorescence and TEM immunogold approaches, we find that SelT is expressed at the endoplasmic reticulum membrane in all hormone-producing pituitary cell types. SelT knockdown in corticotrope cells promotes unfolded protein response (UPR) and ER stress and lowers endoplasmic reticulum-associated protein degradation (ERAD) and hormone production...
September 19, 2017: EMBO Reports
https://www.readbyqxmd.com/read/28923586/genetic-code-asymmetry-supports-diversity-through-experimentation-with-posttranslational-modifications
#8
REVIEW
James W Dennis
Protein N-glycosylation has been identified in all three domains of life presumably conserved for its early role in glycoprotein folding. However, the N-glycans added to proteins in the secretory pathway of multicellular organisms are remodeling in the Golgi, increasing structural diversity exponentially and adding new layers of functionality in immunity, metabolism and other systems. The branching and elongation of N-glycan chains found on cell surface receptors generates a gradation of affinities for carbohydrate-binding proteins, the galectin, selectin and siglec families...
September 15, 2017: Current Opinion in Chemical Biology
https://www.readbyqxmd.com/read/28922188/new-insights-into-the-role-of-glycosylation-in-lipoprotein-metabolism
#9
Marjolein A W van den Boogert, Daniel J Rader, Adriaan G Holleboom
PURPOSE OF REVIEW: Human genetics has provided new insights into the role of protein glycosylation in regulating lipoprotein metabolism. Here we review these new developments and discuss the biological insights they provide. RECENT FINDINGS: Case descriptions of patients with congenital defects in N-glycosylation (CDG-I) frequently describe a distinct hypocholesterolemia in these rare multisystem clinical syndromes. Two novel CDGs with disturbed Golgi homeostasis and trafficking defects result in mixed glycosylation disorders, hepatic steatosis and hypercholesterolemia...
September 15, 2017: Current Opinion in Lipidology
https://www.readbyqxmd.com/read/28920454/investigation-of-o-glycosylation-heterogeneity-of-recombinant-coagulation-factor-ix-using-lc-ms-ms
#10
Youngsuk Seo, Gyeong Mi Park, Myung Jin Oh, Hyun Joo An
AIM: Recombinant coagulation factor IX (rFIX) has extraordinarily multiple post-translational modifications including N-glycosylation and O-glycosylation which have a drastic effect on biological functions and in vivo recovery. Unlike N-glycosylation extensively characterized, there are a few studies on O-glycosylation due to its intrinsic complexity. In-depth O-glycosylation analysis is necessary to better understand and assess pharmacological activity of rFIX. RESULTS: We determined unusual O-glycosylations including O-fucosylation and O-glucosylation which were located at Serine 53 and 61, respectively in EGF domain...
September 18, 2017: Bioanalysis
https://www.readbyqxmd.com/read/28916753/automated-n-glycosylation-sequencing-of-biopharmaceuticals-by-capillary-electrophoresis
#11
Marton Szigeti, Andras Guttman
Comprehensive analysis of the N-linked carbohydrates of glycoproteins is gaining high recent interest in both the biopharmaceutical and biomedical fields. In addition to high resolution glycosylation profiling, sugar residue and linkage specific enzymes are also routinely used for exoglycosidase digestion based carbohydrate sequencing. This latter one, albeit introduced decades ago, still mostly practiced by following tedious and time consuming manual processes. In this paper we introduce an automated carbohydrate sequencing approach using the appropriate exoglycosidase enzymes in conjunction with the utilization of some of the features of a capillary electrophoresis (CE) instrument to speed up the process...
September 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28906588/surface-glycan-pattern-of-canine-equine-and-ovine-bone-marrow-derived-mesenchymal-stem-cells
#12
Salvatore Desantis, Gianluca Accogli, Antonio Crovace, Edda G Francioso, Alberto Maria Crovace
The use of bone marrow-derived mesenchymal stem cells (MSCs) for clinical and experimental studies is increasing, but full characterization of MSCs in veterinary species is hindered by the variability in species-specific cell surface marker expression and antibody cross reactivity. Recent studies demonstrated that the glycans in the glycocalyx of MSCs are promising candidates as cell biomarkers. In the present study, we analyzed the glycocalyx of canine MSCs (cMSCs), ovine MSCs (oMSCs), and equine MSCs (eMSCs) using a cell microarray procedure in which MSCs were spotted on microarray slides and incubated with a panel of 14 biotinylated lectins and Cy3-conjugated streptavidin...
September 14, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
https://www.readbyqxmd.com/read/28900186/improvement-of-the-glycoproteomic-toolbox-with-the-discovery-of-a-unique-c-terminal-cleavage-specificity-of-flavastacin-for-n-glycosylated-asparagine
#13
Alexander Pralow, Marcus Hoffmann, Terry Nguyen-Khuong, Erdmann Rapp, Udo Reichl
To determine all potential N-glycosylation sites of a glycoprotein, one central aspect of every bottom-up N-glycoproteomic strategy is to generate suitable N-glycopeptides that can be detected and analyzed by mass spectrometry. Specific proteases, such as trypsin, bear the potential of generating N-glycopeptides that either carry more than one N-glycosylation site or are too long to be readily analyzed by mass spectrometry- both due to the lack of tryptic cleavage sites near the N-glycosylation site. Here, we present a newly identified cleavage specificity of flavastacin, a protease from Flavobacterium menigosepticum, which - up to now - was only reported to cleave peptide bonds N-terminal to aspartic acid residues...
September 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28898745/glycosylation-is-important-for-fcxth1-activity-as-judged-by-its-structural-and-biochemical-characterization
#14
Ángela Méndez-Yañez, Dina Beltrán, Constanza Campano-Romero, Sebastián Molinett, Raúl Herrera, María Alejandra Moya-León, Luis Morales-Quintana
Xyloglucan endotransglycosylase/hydrolases (XTH) may have endotransglycosylase (XET) and/or hydrolase (XEH) activities. Previous studies suggest that XTHs might play a key role in ripening of Fragaria chiloensis fruit as FcXTH1 transcripts increase as fruit softens. FcXTH1 protein sequence contains a conserved N-glycosylation site adjacent to catalytic residues. The FcXTH1 structure was built through comparative modeling methodology, the structure displays a β-jellyroll-type folding with a curvature generated by eight antiparallel β-sheets that holds the catalytic motif that is oriented towards the central cavity of the protein...
September 9, 2017: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/28895363/-characterization-of-n-glycosylation-in-an-anti-egfr-monoclonal-antibody-produced-by-different-expression-systems
#15
Chong Wang, Huaizu Guo
The use of mammalian expression systems results in a remarkable heterogeneity of mAb products, generally due to post-translational modifications, and glycosylation is a critical post-translation modification because it has a profound impact on the safety and efficacy of mAbs. The present study was designed to explore the impact of a different expression system on mAb N-glycosylation. The detailed structures of individual glycans between anti-EGFR monoclonal antibodies produced by different expression systems were successfully characterized at the level of free oligosaccharides using liquid chromatography electrospray ionization quadrupole time-of-fight mass spectrometry (LC-ESI-QTof MS)...
June 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28892050/carcinoembryonic-antigen-related-cell-adhesion-molecule-6-ceacam6-promotes-egf-receptor-signaling-of-oral-squamous-cell-carcinoma-metastasis-via-the-complex-n-glycosylation
#16
W-F Chiang, T-M Cheng, C-C Chang, S-H Pan, C A Changou, T-H Chang, K-H Lee, S-Y Wu, Y-F Chen, K-H Chuang, D-B Shieh, Y-L Chen, C-C Tu, W-L Tsui, M-H Wu
Aberrant protein glycosylation could be a distinct surface-marker of cancer cells that influences cancer progression and metastasis because glycosylation can regulate membrane protein folding which alters receptor activation and changes epitope exposure for antibody (Ab) recognition. Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), a glycophosphoinositol-anchored protein, is a heavily glycosylated tumor antigen. However, the clinical significance and biological effect of CEACAM6 glycosylation has not been addressed in cancers...
September 11, 2017: Oncogene
https://www.readbyqxmd.com/read/28891400/glycosylated-cell-surface-markers-for-the-isolation-of-human-cardiac-progenitors
#17
Asja T Moerkamp, Hau Wan Leung, Noortje A M Bax, Stephanie Holst, Kirsten Lodder, Thijs Berends, Calinda K E Dingenouts, Andre Boon Hwa Choo, Anke M Smits, Marie-Jose Goumans
The aim of stem cell therapy after cardiac injury is to replace damaged cardiac tissue. Human cardiac progenitor cells (CPCs) represent an interesting cell population for clinical strategies to treat cardiac disease and human CPC-specific antibodies would aid in the clinical implementation of cardiac progenitor based cell therapy. However, the field of CPC biology suffers from the lack of human CPC-specific markers. Therefore, we raised a panel of monoclonal antibodies (mAb) against CPCs Of this panel of antibodies, we show that mAb C1096 recognizes a progenitor-like population in the fetal and adult human heart and partially co-localize with reported CPC populations in vitro...
September 11, 2017: Stem Cells and Development
https://www.readbyqxmd.com/read/28890404/structural-and-functional-characterization-of-a-highly-secreted-%C3%AE-l-arabinofuranosidase-gh62-from-aspergillus-nidulans-grown-on-sugarcane-bagasse
#18
Fabiano Jares Contesini, Marcelo Vizoná Liberato, Marcelo Ventura Rubio, Felipe Calzado, Mariane Paludetti Zubieta, Diego Mauricio Riaño Pachón, Fabio Marcio Squina, Fabricio Bracht, Munir S Skaf, André Ricardo Damásio
Carbohydrate-Active Enzymes are key enzymes for biomass-to-bioproducts conversion. α-L-Arabinofuranosidases that belong to the Glycoside Hydrolase family 62 (GH62) have important applications in biofuel production from plant biomass by hydrolyzing arabinoxylans, found in both the primary and secondary cell walls of plants. In this work, we identified a GH62 α-L-arabinofuranosidase (AnAbf62Awt) that was highly secreted when Aspergillus nidulans was cultivated on sugarcane bagasse. The gene AN7908 was cloned and transformed in A...
September 7, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28890400/deglycosylating-enzymes-acting-on-n-glycans-in-fungi-insights-from-a-genome-survey
#19
REVIEW
Georgios Tzelepis, Magnus Karlsson, Tadashi Suzuki
BACKGROUND: N-Glycosylation, one of the most prominent post-translational modifications of proteins, is found in all domains of life, i.e. eukaryotes, bacteria and archaea, and has been shown to play a crucial role in modulating the physicochemical/physiological properties of carrier proteins. Deglycosylating enzymes that act on N-glycans are widely used in analyzing the structures/functions of N-glycans. Fungi are known to produce various deglycosylating enzymes, some of which are fungi-specific...
September 7, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28887103/unusual-glycosylation-of-proteins-beyond-the-universal-sequon-and-other-amino-acids
#20
REVIEW
Devawati Dutta, Chhabinath Mandal, Chitra Mandal
BACKGROUND: Glycosylation of proteins is the most common, multifaceted co- and post-translational modification responsible for many biological processes and cellular functions. Significant alterations and aberrations of these processes are related to various pathological conditions, and often turn out to be disease biomarkers. Conventional N-glycosylation occurs through the recognition of the consensus sequon, asparagine (Asn)-X-serine (Ser)/threonine (Thr), where X is any amino acid except for proline, with N-acetylglucosamine (GlcNAc) as the first glycosidic linkage...
September 5, 2017: Biochimica et Biophysica Acta
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