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N-glycosylation

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https://www.readbyqxmd.com/read/27917827/the-n14-anti-afamin-antibody-fab-a-rare-vl1-cdr-glycosylation-crystallographic-re-sequencing-molecular-plasticity-and-conservative-versus-enthusiastic-modelling
#1
Andreas Naschberger, Barbara G Fürnrohr, Tihana Lenac Rovis, Suzana Malic, Klaus Scheffzek, Hans Dieplinger, Bernhard Rupp
The monoclonal antibody N14 is used as a detection antibody in ELISA kits for the human glycoprotein afamin, a member of the albumin family, which has recently gained interest in the capture and stabilization of Wnt signalling proteins, and for its role in metabolic syndrome and papillary thyroid carcinoma. As a rare occurrence, the N14 Fab is N-glycosylated at Asn26L at the onset of the VL1 antigen-binding loop, with the α-1-6 core fucosylated complex glycan facing out of the L1 complementarity-determining region...
December 1, 2016: Acta Crystallographica. Section D, Structural Biology
https://www.readbyqxmd.com/read/27914120/effect-of-glycosylation-on-the-biocatalytic-properties-of-hydroxynitrile-lyase-from-the-passion-fruit-passiflora-edulis-a-comparison-of-natural-and-recombinant-enzymes
#2
Aem Nuylert, Yuko Ishida, Yasuhisa Asano
A hydroxynitrile lyase from the passion fruit, Passiflora edulis (PeHNL) was isolated from the leaves and showed high stability in biphasic co-organic solvent systems for cyanohydrin synthesis. Cyanohydrins are important building blocks for the production of fine chemicals and pharmaceuticals. Thus, to enhance production yields of PeHNL for industrial applications, we cloned and expressed recombinant PeHNL in E. coli BL21 (DE3) and P. pastoris GS115 cells without a signal peptide sequence. The aim of the present study is to determine the effect of N-glycosylation on enzyme stability and catalytic properties in microbial expression systems...
December 3, 2016: Chembiochem: a European Journal of Chemical Biology
https://www.readbyqxmd.com/read/27912985/immunogenicity-and-malaria-transmission-reducing-potency-of-pfs48-45-and-pfs25-encoded-by-dna-vaccines-administered-by-intramuscular-electroporation
#3
Dibyadyuti Datta, Geetha P Bansal, Dietlind L Gerloff, Barry Ellefsen, Drew Hannaman, Nirbhay Kumar
Pfs48/45 and Pfs25 are leading candidates for the development of Plasmodium falciparum transmission blocking vaccines (TBV). Expression of Pfs48/45 in the erythrocytic sexual stages and presentation to the immune system during infection in the human host also makes it ideal for natural boosting. However, it has been challenging to produce a fully folded, functionally active Pfs48/45, using various protein expression platforms. In this study, we demonstrate that full-length Pfs48/45 encoded by DNA plasmids is able to induce significant transmission reducing immune responses...
November 29, 2016: Vaccine
https://www.readbyqxmd.com/read/27911384/analysis-of-scap-n-glycosylation-and-trafficking-in-human-cells
#4
Chunming Cheng, Jeffrey Yunhua Guo, Feng Geng, Xiaoning Wu, Xiang Cheng, Qiyue Li, Deliang Guo
Elevated lipogenesis is a common characteristic of cancer and metabolic diseases. Sterol regulatory element-binding proteins (SREBPs), a family of membrane-bound transcription factors controlling the expression of genes important for the synthesis of cholesterol, fatty acids and phospholipids, are frequently upregulated in these diseases. In the process of SREBP nuclear translocation, SREBP-cleavage activating protein (SCAP) plays a central role in the trafficking of SREBP from the endoplasmic reticulum (ER) to the Golgi and in subsequent proteolysis activation...
November 8, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27908797/molecular-detection-and-characterization-of-respiratory-syncytial-virus-b-genotypes-circulating-in-pakistani-children
#5
Uzma Bashir, Nadia Nisar, Nayab Mahmood, Muhammad Masroor Alam, Hajra Sadia, Syed Sohail Zahoor Zaidi
Respiratory syncytial virus (RSV) is the major cause of acute lower respiratory tract infections in young children, but very little is known about its epidemiology and circulating genotypes in Pakistan. This study analyzed the epidemiological and molecular characteristics of RSV B genotypes in Pakistani children below 5years with acute respiratory tract infections (ARIs) during three consecutive winter seasons from 2010 to 2013. A total of 1941 samples were analyzed for RSV infection by real time PCR and 24% (472/1941) samples were found positive out of which 22...
November 28, 2016: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/27908295/dopamine-2-receptor-extracellular-n-terminus-regulates-receptor-surface-availability-and-is-the-target-of-human-pathogenic-antibodies-from-children-with-movement-and-psychiatric-disorders
#6
Nese Sinmaz, Fiona Tea, Deepti Pilli, Alicia Zou, Mazen Amatoury, Tina Nguyen, Vera Merheb, Sudarshini Ramanathan, Sandra T Cooper, Russell C Dale, Fabienne Brilot
Anti-Dopamine-2 receptor (D2R) antibodies have been recently identified in a subgroup of children with autoimmune movement and psychiatric disorders, however the epitope(s) and mechanism of pathogenicity remain unknown. Here we report a major biological role for D2R extracellular N-terminus as a regulator of receptor surface availability, and as a major epitope targeted and impaired in brain autoimmunity. In transfected human cells, purified anti-D2R antibody from patients specifically and significantly reduced human D2R surface levels...
December 1, 2016: Acta Neuropathologica Communications
https://www.readbyqxmd.com/read/27902333/plant-produced-anti-dengue-virus-monoclonal-antibodies-exhibit-reduced-antibody-dependent-enhancement-of-infection-activity
#7
Matthew Dent, Jonathan Hurtado, Amber M Paul, Haiyan Sun, Huafang Lai, Ming Yang, Adrian Esqueda, Fengwei Bai, Herta Steinkellner, Qiang Chen
The monoclonal antibody (mAb) E60 has the potential to be a desirable therapeutic molecule since it efficiently neutralizes all four serotypes of dengue virus (DENV). However, mammalian cell-produced E60 exhibits antibody-dependent enhancement of infection (ADE) activity, rendering it inefficacious in vivo and subjects treated animals more susceptible to develop more severe diseases during secondary infection. In this study, we evaluated a plant-based expression system for the production of therapeutically suitable E60...
October 20, 2016: Journal of General Virology
https://www.readbyqxmd.com/read/27896447/novel-extended-sequons-of-human-n-glycosylation-sites-improve-the-precision-of-qualitative-predictions-an-alignment-free-study-of-pattern-recognition-using-protdcal-protein-features
#8
Yasser B Ruiz-Blanco, Yovani Marrero-Ponce, Enrique García-Hernández, James Green
N-Glycosylation is a common post-translational modification that plays an important role in the proper folding and function of many proteins. This modification is largely dependent on the presence of a sequence motif called a "sequon" defined as Asn-Xxx-Ser/Thr. However, evidence has shown that the presence of such a "sequon" is insufficient to determine the occurrence of N-glycosylation with high precision. This study aims to elucidate patterns that can more accurately predict N-glycosylation sites in human proteins...
November 28, 2016: Amino Acids
https://www.readbyqxmd.com/read/27895582/introduction-of-an-n-glycosylation-site-into-udp-glucuronosyltransferase-2b3-alters-its-sensitivity-to-cytochrome-p450-3a1-dependent-modulation
#9
Tatsuro Nakamura, Naho Yamaguchi, Yuu Miyauchi, Tomoki Takeda, Yasushi Yamazoe, Kiyoshi Nagata, Peter I Mackenzie, Hideyuki Yamada, Yuji Ishii
Our previous studies have demonstrated functional protein-protein interactions between cytochrome P450 (CYP) 3A and UDP-glucuronosyltransferase (UGT). However, the role of carbohydrate chains of UGTs in the interaction with CYP is not well understood. To address this issue, we examined whether CYP3A1 modulates the function of UGT2B3 which lacks potential glycosylation sites. We also examined whether the introduction of N-glycosylation to UGT2B3 affects CYP3A-dependent modulation of UGT function. To introduce a potential glycosylation site into UGT2B3, Ser 316 of UGT2B3 was substituted with Asn by site-directed mutagenesis...
2016: Frontiers in Pharmacology
https://www.readbyqxmd.com/read/27887626/expression-and-processing-analyses-of-wild-type-and-p-r47h-trem2-variant-in-alzheimer-s-disease-brains
#10
Li Ma, Mariet Allen, Nobutaka Sakae, Nilufer Ertekin-Taner, Neill R Graff-Radford, Dennis W Dickson, Steven G Younkin, Daniel Sevlever
BACKGROUND: Genetic analyses showed that the triggering receptor expressed in myeloid cells 2 (TREM2) p.R47H variant increases the risk for Alzheimer's disease (AD). The question of whether the p.R47H mutation affects expression or function of the receptor remains unanswered. To address this question we quantified mRNA and analyzed protein profiles of WT and p.R47H TREM2 in human brains. METHODS: Quantitative real-time PCR (qPCR) was performed using 2 sets of primers one that detects all TREM2 mRNA isoforms and one specific for the alternative spliced isoform (TREM2alt) that encodes for the extracellular domain (soluble TREM2)...
November 25, 2016: Molecular Neurodegeneration
https://www.readbyqxmd.com/read/27869218/n-linked-glycosylation-at-asn152-on-cd147-affects-protein-folding-and-stability-promoting-tumour-metastasis-in-hepatocellular-carcinoma
#11
Jiang-Hua Li, Wan Huang, Peng Lin, Bo Wu, Zhi-Guang Fu, Hao-Miao Shen, Lin Jing, Zhen-Yu Liu, Yang Zhou, Yao Meng, Bao-Qing Xu, Zhi-Nan Chen, Jian-Li Jiang
Cluster of differentiation 147 (CD147), also known as extracellular matrix metalloproteinase inducer, is a transmembrane glycoprotein that mediates oncogenic processes partly through N-glycosylation modifications. N-glycosylation has been demonstrated to be instrumental for the regulation of CD147 function during malignant transformation. However, the role that site-specific glycosylation of CD147 plays in its defective function in hepatocellular carcinomacells needs to be determined. Here, we demonstrate that the modification of N-glycosylation at Asn152 on CD147 strongly promotes hepatocellular carcinoma (HCC) invasion and migration...
November 21, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27868074/cd63-promotes-hemocyte-mediated-phagocytosis-in-the-clam-paphia-undulata
#12
Mingjia Yu, Shanjun Yang, Hongxia Sun, Qiang Xia
As one of the surface membrane proteins of tetraspanin family, CD63 plays a crucial role in cellular trafficking and endocytosis, which also is associated with activation of a wide variety of immune cells. Here, the homolog of CD63 was characterized from one marine mollusk, Paphia undulata, which is designated as Pu-CD63. The complete cDNA of Pu-CD63 is 1,738 bp in length with an open reading frame (ORF) of 849 bp, encoding a 282 amino acid protein with four putative hydrophobic transmembrane helixes. Bioinformatic analysis revealed that Pu-CD63 contains one putative YXXØ consensus motif of "110-YVII-113" and one N-glycosylation site "155-NGT-157" within the large extracellular loop (LEL) region, supporting its conserved function in plasma membrane and endosomal/lysosomal trafficking...
2016: Journal of Immunology Research
https://www.readbyqxmd.com/read/27865927/structural-perturbations-induced-by-asn131-and-asn171-glycosylation-converge-within-the-efsam-core-and-enhance-stromal-interaction-molecule-1-mediated-store-operated-calcium-entry
#13
Yoo Jung Choi, Yue Zhao, Moshmi Bhattacharya, Peter B Stathopulos
A major intracellular calcium (Ca(2+)) uptake pathway in both excitable and non-excitable eukaryotic cells is store-operated Ca(2+) entry (SOCE). SOCE is the process by which endoplasmic reticulum (ER)-stored Ca(2+) depletion leads to activation of plasma membrane Ca(2+) channels to provide a sustained increase in cytosolic Ca(2+) levels that mediate a plethora of physiological processes ranging from the immune response to platelet aggregation. Stromal interaction molecule-1 (STIM1) is the principal regulator of SOCE and responds to changes in ER stored Ca(2+) through luminal sensing machinery composed of EF-hand and SAM domains (EFSAM)...
November 17, 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27864899/n-glycosylation-at-asn-402-stabilizes-n-cadherin-and-promotes-cell-cell-adhesion-of-glioma-cells
#14
Yaolin Xu, Ruiqi Chang, Fulin Xu, Yan Gao, Fuming Yang, Can Wang, Jin Xiao, Zuopeng Su, Yongyan Bi, Liying Wang, Xiliang Zha
Cadherin is crucial for cell-cell adhesion and N-glycosylation of N-cadherin has been implicated in the process of mammary, renal, and ovarian carcinogenesis. However, whether N-glycosylation of N-cadherin plays a role in glioma remains unknown. Previous studies had indicated that N-glycosylation could occur at three asparagine residues of N-cadherin. By generating and over-expressing N-glycosylation-deficient N-cadherin mutants in the human glioma cell lines SHG66 and U87, we found that mutation of N402 but not of the other potentially N-glycosylated residues destabilized N-cadherin and led to its ubiquitylation and subsequent proteasomal degradation...
November 19, 2016: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/27864768/hormone-receptor-glycosylation
#15
Ulrike Vavra, Christiane Veit, Richard Strasser
Glycosylation is essential for all trees of life. N-glycosylation is one of the most common covalent protein modifications and influences a large variety of cellular processes including protein folding, quality control and protein-receptor interactions. Despite recent progress in understanding of N-glycan biosynthesis, our knowledge of N-glycan function on individual plant proteins is still very limited. In this respect, plant hormone receptors are an interesting group of proteins as several of these proteins are present at distinct sites in the secretory pathway or at the plasma membrane and have numerous potential N-glycosylation sites...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27855667/molecular-evolution-of-type-2-porcine-reproductive-and-respiratory-syndrome-viruses-circulating-in-vietnam-from-2007-to-2015
#16
Hai Quynh Do, Dinh Thau Trinh, Thi Lan Nguyen, Thi Thu Hang Vu, Duc Duong Than, Thi Van Lo, Minjoo Yeom, Daesub Song, SeEun Choe, Dong-Jun An, Van Phan Le
BACKGROUND: Porcine respiratory and reproductive syndrome (PRRS) virus is one of the most economically significant pathogens in the Vietnamese swine industry. ORF5, which participates in many functional processes, including virion assembly, entry of the virus into the host cell, and viral adaptation to the host immune response, has been widely used in molecular evolution and phylogeny studies. Knowing of molecular evolution of PRRSV fields strains might contribute to PRRS control in Vietnam...
November 17, 2016: BMC Veterinary Research
https://www.readbyqxmd.com/read/27849032/the-disruption-of-gdp-fucose-de-novo-biosynthesis-suggests-the-presence-of-a-novel-fucose-containing-glycoconjugate-in-plasmodium-asexual-blood-stages
#17
Sílvia Sanz, Borja López-Gutiérrez, Giulia Bandini, Sebastian Damerow, Sabrina Absalon, Rhoel R Dinglasan, John Samuelson, Luis Izquierdo
Glycosylation is an important posttranslational protein modification in all eukaryotes. Besides glycosylphosphatidylinositol (GPI) anchors and N-glycosylation, O-fucosylation has been recently reported in key sporozoite proteins of the malaria parasite. Previous analyses showed the presence of GDP-fucose (GDP-Fuc), the precursor for all fucosylation reactions, in the blood stages of Plasmodium falciparum. The GDP-Fuc de novo pathway, which requires the action of GDP-mannose 4,6-dehydratase (GMD) and GDP-L-fucose synthase (FS), is conserved in the parasite genome, but the importance of fucose metabolism for the parasite is unknown...
November 16, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27834568/the-tgf-%C3%AE-inhibitory-activity-of-antibody-37e1b5-depends-on-its-h-cdr2-glycan
#18
Ping Tsui, Daniel R Higazi, Yanli Wu, Rebecca Dunmore, Emilie Solier, Toyin Kasali, Nicholas J Bond, Catherine Huntington, Alan Carruthers, John Hood, M Jack Borrok, Arnita Barnes, Keith Rickert, Sandrina Phipps, Lena Shirinian, Jie Zhu, Michael A Bowen, William Dall'Acqua, Lynne A Murray
Excessive transforming growth factor (TGF)-β is associated with pro-fibrotic responses in lung disease, yet it also plays essential roles in tissue homeostasis and autoimmunity. Therefore, selective inhibition of excessive and aberrant integrin-mediated TGF-β activation via targeting the alpha-v family of integrins is being pursued as a therapeutic strategy for chronic lung diseases, to mitigate any potential safety concerns with global TGF-β inhibition. In this work, we reveal a novel mechanism of inhibiting TGF-β activation utilized by an αvβ8 targeting antibody, 37E1B5...
November 11, 2016: MAbs
https://www.readbyqxmd.com/read/27832592/receptor-mediated-sorting-of-typhoid-toxin-during-its-export-from-salmonella-typhi-infected-cells
#19
Shu-Jung Chang, Jeongmin Song, Jorge E Galán
Typhoid toxin is an essential virulence factor of Salmonella Typhi, the cause of typhoid fever. Typhoid toxin is secreted into the lumen of Salmonella-containing vacuole (SCV), after which it is packaged into vesicle carrier intermediates and released extracellularly through incompletely understood mechanisms. Following export, the toxin targets cells by interacting with human-specific Neu5Ac-terminated glycan receptors. We show that typhoid toxin is sorted from the SCV into vesicle carrier intermediates via interactions of its B subunit, PltB, with specific lumenal sialylated glycan packaging receptors...
November 9, 2016: Cell Host & Microbe
https://www.readbyqxmd.com/read/27822701/aglh-a-thermophilic-udp-n-acetylglucosamine-1-phosphate-dolichyl-phosphate-glcnac-1-phosphotransferase-initiating-protein-n-glycosylation-pathway-in-sulfolobus-acidocaldarius-is-capable-of-complementing-the-eukaryal-alg7
#20
Benjamin H Meyer, Hosam Shams-Eldin, Sonja-Verena Albers
AglH, a predicted UDP-GlcNAc-1-phosphate:dolichyl phosphate GlcNAc-1-phosphotransferase, is initiating the protein N-glycosylation pathway in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius. AglH successfully replaced the endogenous GlcNAc-1-phosphotransferase activity of Alg7 in a conditional lethal Saccharomyces cerevisiae strain, in which the first step of the eukaryal protein N-glycosylation process was repressed. This study is one of the few examples of cross-domain complementation demonstrating a conserved polyprenyl phosphate transferase reaction within the eukaryal and archaeal domain like it was demonstrated for Methanococcus voltae (Shams-Eldin et al...
November 7, 2016: Extremophiles: Life Under Extreme Conditions
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