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N-glycosylation

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https://www.readbyqxmd.com/read/29351237/bioengineering-of-rfviia-biopharmaceutical-and-structure-characterization-for-biosimilarity-assessment
#1
Othman Montacir, Houda Montacir, Murat Eravci, Andreas Springer, Stephan Hinderlich, Fereidoun Mahboudi, Amirhossein Saadati, Maria Kristina Parr
Eptacog alfa (NovoSeven®) is a vitamin K-dependent recombinant Factor VIIa produced by genetic engineering from baby hamster kidney (BHK) cells as a single peptide chain of 406 residues. After activation, it consists of a light chain (LC) of 152 amino and a heavy chain (HC) of 254 amino acids. Recombinant FVIIa undergoes many post-translational modifications (PTMs). The first ten glutamic acids of the N-terminal moiety are γ-carboxylated, Asn145 and Asn322 are N-glycosylated, and Ser52 and Ser60 are O-glycosylated...
January 19, 2018: Bioengineering
https://www.readbyqxmd.com/read/29346724/structural-and-biophysical-characterization-of-human-extl3-domain-organisation-glycosylation-and-solution-structure
#2
Wael Awad, Sven Kjellstrom, Gabriel Svensson Birkedal, Katrin Mani, Derek Thomas Logan
Heparan sulfate proteoglycans are proteins substituted with one or more heparan sulfate (HS) polysaccharides, found in abundance at cell surfaces. HS chains influence the activity of many biologically important molecules involved in cellular communication and signaling. The exostosin (EXT) proteins are glycosyltransferases in the Golgi apparatus that assemble HS chains on HSPGs. The EXTL3 enzyme mainly works as an initiator in HS biosynthesis. In this work, human lumenal N-glycosylated EXTL3 (EXTL3ΔN) was cloned, expressed in human embryonic kidney cells and purified...
January 18, 2018: Biochemistry
https://www.readbyqxmd.com/read/29345911/role-of-glycanation-and-convertase-maturation-of-the-soluble-glypican-3-in-inhibiting-proliferation-of-hepatocellular-carcinoma-cells
#3
Ahmad Saad, Benjamin Liet, Gilles Joucla, Xavier Santarelli, Justine Charpentier, Stéphane Claverol, Christophe F Grosset, Véronique Trézéguet
Glypican 3 (GPC3) is a complex heparan sulfate proteoglycan associated with the outer surface of the plasma membrane by a glycosyl-phosphatidylinositol anchor (GPI). It is also N-glycosylated and processed by a furine-like convertase. GPC3 has numerous biological functions. While undetectable in normal liver tissue, it is abnormally and highly overexpressed in hepatocellular carcinoma (HCC). Interestingly, proliferation of HCC cells such as HepG2 and HuH7 is inhibited when they express a soluble form of GPC3 after lentiviral transduction...
January 18, 2018: Biochemistry
https://www.readbyqxmd.com/read/29343697/effluent-and-serum-protein-n-glycosylation-is-associated-with-inflammation-and-peritoneal-membrane-transport-characteristics-in-peritoneal-dialysis-patients
#4
Evelina Ferrantelli, Karima Farhat, Agnes L Hipgrave Ederveen, Karli R Reiding, Robert H J Beelen, Frans J van Ittersum, Manfred Wuhrer, Viktoria Dotz
Mass spectrometric glycomics was used as an innovative approach to identify biomarkers in serum and dialysate samples from peritoneal dialysis (PD) patients. PD is a life-saving treatment worldwide applied in more than 100,000 patients suffering from chronic kidney disease. PD treatment uses the peritoneum as a natural membrane to exchange waste products from blood to a glucose-based solution. Daily exposure of the peritoneal membrane to these solutions may cause complications such as peritonitis, fibrosis and inflammation which, in the long term, lead to the failure of the treatment...
January 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29343654/intestinal-epithelial-cell-specific-deletion-of-%C3%AE-mannosidase-ii-ameliorates-experimental-colitis
#5
Koichiro Suzuki, Takahiro Yamada, Keiko Yamazaki, Masato Hirota, Narumi Ishihara, Mizuki Sakamoto, Daisuke Takahashi, Hideki Iijima, Koji Hase
Inflammatory bowel disease (IBD) is a refractory disease of the gastrointestinal tract that is believed to develop in genetically susceptible individuals. Glycosylation, a type of post-translational modification, is involved in the development of a wide range of diseases, including IBD, by modulating the function of various glycoproteins. To identify novel genes contributing to the development of IBD, we analyzed single nucleotide polymorphisms (SNPs) of glycosylation-related genes in IBD patients and identified MAN2A1, encoding alpha-mannosidase II (α-MII), as a candidate gene...
January 18, 2018: Cell Structure and Function
https://www.readbyqxmd.com/read/29330871/multi-site-n-glycan-mapping-study-2-uhplc
#6
Ákos Szekrényes, SungAe Suhr Park, Eoin Cosgrave, Aled Jones, Ted Haxo, Michael Kimzey, Shiva Pourkaveh, Zoltán Szabó, Zoran Sosic, Peng Feng, Preeti Sejwal, Kelsey Dent, David Michels, Gordon Freckleton, Jun Qian, Catherine Lancaster, Toni Duffy, Melissa Schwartz, Jiann-Kae Luo, Jonathan van Dyck, Pui-King Leung, Marcell Olajos, Ronald Kowle, Kai Gao, Wenbo Wang, Jo Wegstein, Samnang Tep, Apolka Domokos, Csaba Váradi, András Guttman
In the first part of this publication, the results from an international study evaluating the precision (i.e., repeatability and reproducibility) of N-glycosylation analysis using capillary electrophoresis of APTS-labeled N-glycans were presented. The corresponding results from ultra-high performance liquid chromatography (UHPLC) with fluorescence detection are presented here from 12 participating sites. All participants used the same lot of samples, reagents, and columns to perform the assays. Elution time, peak area and peak area percent values were determined for all peaks ≥0...
January 13, 2018: Electrophoresis
https://www.readbyqxmd.com/read/29329339/accessibility-and-contribution-to-glucan-masking-of-natural-and-genetically-tagged-versions-of-yeast-wall-protein-1-of-candida-albicans
#7
Bruce L Granger
Yeast wall protein 1 (Ywp1) is an abundant glycoprotein of the cell wall of the yeast form of Candida albicans, the most prevalent fungal pathogen of humans. Antibodies that bind to the polypeptide backbone of isolated Ywp1 show little binding to intact yeast cells, presumably because the Ywp1 epitopes are masked by the polysaccharides of the mannoproteins that form the outer layer of the cell wall. Rare cells do exhibit much greater anti-Ywp1 binding, however, and one of these was isolated and characterized...
2018: PloS One
https://www.readbyqxmd.com/read/29329315/role-of-n-glycosylation-in-activation-of-prommp-9-a-molecular-dynamics-simulations-study
#8
Sonu Kumar, Piotr Cieplak
Human matrix metalloproteinase proMMP-9 is secreted as latent zymogen, which requires two-steps proteolytic activation. The secreted proMMP-9 is glycosylated at two positions: Asn38 and Asn120 located in the prodomain and catalytic domain, respectively. It has been demonstrated that glycosylation at Asn120 is required for secretion of the enzyme, while the role of Asn38 glycosylation is not well understood, but is usually linked to the activation process. One hypothesis stated that the Asn38 glycosylation might protect against proteolytic activation...
2018: PloS One
https://www.readbyqxmd.com/read/29328676/stabilizing-leaf-and-branch-compost-cutinase-lcc-with-glycosylation-mechanism-and-effect-on-pet-hydrolysis
#9
Abhijit N Shirke, Christine White, Jacob A Englaender, Allison Sophie Zwarycz, Glenn L Butterfoss, Robert J Linhardt, Richard A Gross
Cutinases are polyester hydrolases that show a remarkable capability to hydrolyze polyethylene terephthalate (PET) to its monomeric units. This revelation has stimulated research aimed at developing sustainable and green cutinase-catalyzed PET recycling methods. Leaf and branch compost cutinase (LCC) is particularly suited towards these ends given its relatively high PET hydrolysis activity and thermostability. Any practical enzymatic PET recycling application will require that the protein have kinetic stability at or above the PET glass transition temperature (Tg, i...
January 12, 2018: Biochemistry
https://www.readbyqxmd.com/read/29323465/multiplexed-digital-mrna-expression-analysis-profiles-system-wide-changes-in-mrna-abundance-and-responsiveness-of-upr-specific-gene-expression-changes-during-batch-culture-of-recombinant-chinese-hamster-ovary-cells
#10
Rodrigo Maldonado-Agurto, Alan J Dickson
The unfolded protein response (UPR) signalling pathway is viewed as critical for setting the effectiveness of recombinant protein expression in CHO cells. In this study, we used Nanostring nCounter technology to study expression of a group of genes associated with cellular processes linked to UPR activation under ER stress and the changing environment of a batch culture. Time course induction of ER stress, using tunicamycin (TM), showed a group of genes such as Chop, Trb3, Sqstm1, Grp78 and Herpud1 responded rapidly to TM inhibition of N-glycosylation, whilst others such as Atf5, Odz4 and Birc5 exhibited a delayed response...
January 11, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29317657/blood-plasma-igg-n-glycome-biosignatures-associated-with-major-depressive-disorder-symptom-severity-and-the-antidepressant-response
#11
Dong Ik Park, Jerko Štambuk, Genadij Razdorov, Maja Pučić-Baković, Daniel Martins-de-Souza, Gordan Lauc, Christoph W Turck
While N-linked glycosylation has been extensively studied in the context of inflammatory and metabolic disorders, its relationship with major depressive disorder (MDD) and antidepressant treatment response has not been investigated. In our exploratory study, we analysed N-glycan profiles in blood plasma samples collected from MDD patients (n = 18) and found gender-dependent correlations with severity of depressive symptoms prior to initiating antidepressant treatment. In addition, several N-glycosylation traits showed gender-dependent associations with clinical antidepressant response...
January 9, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29315945/inhibitory-effect-of-a-reproductive-related-serpin-on-sperm-trypsin-like-activity-implicates-its-role-in-sperm-maturation-of-penaeus-monodon
#12
Charoonroj Chotwiwatthanakun, Wanida Santimanawong, Prasert Sobhon, Somjai Wongtripop, Rapeepun Vanichviriyakit
In a number of marine animals, sperm serine proteases are important for fertilization. Penaeus monodon sperm require trypsin-like activity for a complete acrosome reaction, which exclusively occurs in sperm residing in the female thelycum. In this study, a complete cDNA sequence of reproductive tract-related Serine protease inhibitor (rrPmserpin) was identified. The longest open reading frame was composed of 1,366 nucleotides encoding 402 amino acids with a predicted pI of 6.86 and molecular mass of 44.88 kDa...
January 5, 2018: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/29311258/molecular-mechanisms-of-cutis-laxa-and-distal-renal-tubular-acidosis-causing-mutations-in-v-atpase-a-subunits-atp6v0a2-and-atp6v0a4
#13
Sally Esmail, Norbert Kartner, Yeqi Yao, Joo Wan Kim, Reinhart A F Reithmeier, Morris F Manolson
The a subunit is the largest of 15 different subunits that make up the vacuolar H+-ATPase (V-ATPase) complex, where it functions in proton translocation. In mammals, this subunit has four paralogous isoforms, a1-a4, which may encode signals for targeting assembled V-ATPases to specific intracellular locations. Despite the functional importance of the a subunit, its structure remains controversial. By studying molecular mechanisms of human disease-causing missense mutations within a subunit isoforms, we may identify domains critical for V-ATPase targeting, activity and/or regulation...
January 8, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29306003/genetic-analysis-of-l1r-myristoylated-protein-of-capripoxviruses-reveals-structural-homogeneity-among-poxviruses
#14
Monu Karki, Amit Kumar, Gnanavel Venkatesan, Sargam Arya, A B Pandey
Sheeppox virus (SPPV) and goatpox virus (GTPV) are members of the genus Capripoxvirus (CaPV) of the family Poxviridae. CaPVs are responsible for important contagious diseases of small ruminants that are enzootic to the Indian sub-continent, Central and Northern Africa and the Middle East. In the present study, the sequence and phylogenetic analysis of the L1R gene of sixteen CaPV isolates (seven SPPV and nine GTPV) from India were performed along with 3D homology modeling of the L1R protein. L1R is a myristoylated protein responsible for virion assembly and being present on intracellular mature virion (IMV) surface, it is also a potent target for eliciting neutralizing antibodies...
January 3, 2018: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/29301962/structure-of-the-yeast-oligosaccharyltransferase-complex-gives-insight-into-eukaryotic-n-glycosylation
#15
Rebekka Wild, Julia Kowal, Jillianne Eyring, Elsy M Ngwa, Markus Aebi, Kaspar P Locher
Oligosaccharyltransferase (OST) is an essential membrane protein complex in the Endoplasmic Reticulum, where it transfers an oligosaccharide from a dolichol-pyrophosphate-activated donor to glycosylation sites of secretory proteins. We here describe the atomic structure of yeast OST determined by cryo-EM, revealing a conserved subunit arrangement. The active site of the catalytic STT3 subunit points away from the center of the complex, allowing unhindered access to substrates. The dolichol-pyrophosphate moiety binds to a lipid-exposed groove of STT3, while two non-catalytic subunits and an ordered N-glycan form a membrane-proximal pocket for the oligosaccharide...
January 4, 2018: Science
https://www.readbyqxmd.com/read/29301397/haloferax-volcanii-proteome-response-to-deletion-of-a-rhomboid-protease-gene
#16
Mariana I Costa, Micaela Cerletti, Roberto A Paggi, Christian Trötschel, Rosana De Castro, Ansgar Poetsch, Maria I Gimenez
Rhomboids are conserved intramembrane serine proteases involved in cell signaling processes. Their role in prokaryotes is scarcely known and remains to be investigated in Archaea. We previously constructed a rhomboid homolog deletion mutant (ΔrhoII) in Haloferax volcanii, which showed reduced motility, increased novobiocin sensitivity and an N- glycosylation defect. To address the impact of rhoII deletion on H. volcanii physiology, the proteomes of mutant and parental strains were compared by shotgun proteomics...
January 5, 2018: Journal of Proteome Research
https://www.readbyqxmd.com/read/29301365/how-sweet-are-our-gut-beneficial-bacteria-a-focus-on-protein-glycosylation-in-lactobacillus
#17
REVIEW
Dimitrios Latousakis, Nathalie Juge
Protein glycosylation is emerging as an important feature in bacteria. Protein glycosylation systems have been reported and studied in many pathogenic bacteria, revealing an important diversity of glycan structures and pathways within and between bacterial species. These systems play key roles in virulence and pathogenicity. More recently, a large number of bacterial proteins have been found to be glycosylated in gut commensal bacteria. We present an overview of bacterial protein glycosylation systems (O- and N-glycosylation) in bacteria, with a focus on glycoproteins from gut commensal bacteria, particularly Lactobacilli...
January 3, 2018: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29288232/n-glycoproteomic-characterization-of-mannosidase-and-xylotransferase-mutant-strains-of-chlamydomonas
#18
Stefan Schulze, Anne Oltmanns, Nick Machnik, Gai Liu, Nannan Xu, Niklas Jarmatz, Martin Scholz, Kazuhiko Sugimoto, Christian Fufezan, Kaiyao Huang, Michael Hippler
At present, only little is known about the enzymatic machinery required for N-glycosylation in Chlamydomonas reinhardtii, leading to the formation of N-glycans harboring xylose (Xyl) and methylated mannose. This machinery possesses new enzymatic features, as C. reinhardtii N-glycans are independent of β1,2-N-acetylglucosaminyltransferase I (GnTI). Here we have performed comparative N-glycoproteomic analyses of insertional mutants of mannosidase 1A (IMMan1A) and xylosyltransferase 1A (IMXylT1A). The disruption of man1A affected methylation of mannoses and the addition of terminal Xyl...
December 29, 2017: Plant Physiology
https://www.readbyqxmd.com/read/29284210/genomic-characterization-and-pathogenic-study-of-two-porcine-reproductive-and-respiratory-syndrome-viruses-with-different-virulence-in-fujian-china
#19
Xiukun Sui, Ting Xin, Xiaoyu Guo, Hong Jia, Ming Li, Xintao Gao, Jing Wu, Yitong Jiang, Willems L, Hongfei Zhu, Shaohua Hou
Two strains of PRRSVs were isolated in 2006 and 2016, respectively, and designated as FZ06A and FZ16A. Through the inoculation experiments, it was shown that FZ06A caused 100% morbidity and 60% mortality, while FZ16A caused 100% morbidity without death. By using genomic sequence and phylogenetic analyses, close relation of Chinese highly pathogenic PRRSV strain with FZ06A and FZ16A strains was achieved. Based on the achieved results, multiple genomic variations in Nsp2, a unique N-glycosylation site (N³³→K³³), and a K151 AA substitution for virulence in the GP5 of FZ16A were detected except of 30 amino acids (AA) deletion in the NSP2-coding region...
December 28, 2017: Journal of Veterinary Science
https://www.readbyqxmd.com/read/29282902/construction-of-green-fluorescence-protein-mutant-to-monitor-stt3b-dependent-n-glycosylation
#20
Toshihiko Kitajima, Wei Xue, Yi-Shi Liu, Chun-Di Wang, Si-Si Liu, Morihisa Fujita, Xiao-Dong Gao
Oligosaccharyltransferases (OSTs) mediate the en bloc transfer of N-glycan intermediates onto the asparagine residue in glycosylation sequons (N-X-S/T, X≠P). These enzymes are typically heteromeric complexes composed of several membrane-associated subunits, in which STT3 is highly conserved as a catalytic core. Metazoan organisms encode two STT3 genes (STT3A and STT3B) in their genome, resulting in the formation of at least two distinct OST isoforms consisting of shared subunits and complex specific subunits...
December 28, 2017: FEBS Journal
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