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yeast glycosylation

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https://www.readbyqxmd.com/read/28155168/expression-and-characteristics-of-an-endoglucanase-from-trichoderma-atroviride-taegii-in-saccharomyces-cerevisiae
#1
Xiao-Mei Huang, Qing-Qing Li, Xiu-Ling Chen, Jin-Xia Fan, Xiu-Hong Xu, Xu-Dong Sun, Dong-Yu Li, Hong-Xiao Zhao
Endoglucanase secreted by the fungus Trichoderma atroviride is a kind of cellulase. An endoglucanase gene egII was cloned from T. atroviride AS3.3013 and expressed in Saccharomyces cerevisiae INVScI. The open reading frame of the egII gene was composed of 1257 bp, encoding 418 amino acids with a molecular weight of 44.23 kDa plus a signal peptide of 21 amino acids. Based on sequence similarity, TaEGII belonged to the glycosyl hydrolase family 5. Expression of the egII gene in T. atroviride AS3.3013 can be induced by microcrystalline cellulose (MCC), bran, carboxymethyl cellulose (CMC), rice straw, and corn stalk but is inhibited by glucose...
February 2, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28154008/eng1-and-exg8-are-the-major-%C3%AE-glucanases-secreted-by-the-fungal-pathogen-histoplasma-capsulatum
#2
Andrew L Garfoot, Kacey L Dearing, Andrew D VanSchoiack, Vicki H Wysocki, Chad A Rappleye
Fungal cell walls contain β-glucan polysaccharides which stimulate immune responses when recognized by host immune cells. The fungal pathogen Histoplasma capsulatum minimizes detection of β-glucan by host cells through at least two mechanisms: concealment of β-glucans beneath α-glucans and enzymatic removal of any exposed β-glucan polysaccharides by the secreted glucanase Eng1. Histoplasma yeasts also secrete the putative glucanase Exg8, which may serve a similar role as Eng1 in removing exposed β-glucans from the yeast cell surface...
February 2, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28114750/acidic-and-uncharged-polar-residues-in-the-consensus-motifs-of-the-yeast-ca-2-transporter-gdt1p-are-required-for-calcium-transport
#3
Anne-Sophie Colinet, Louise Thines, Antoine Deschamps, Gaëlle Flémal, Didier Demaegd, Pierre Morsomme
The UPF0016 family is a recently identified group of poorly characterized membrane proteins whose function is conserved through evolution and that are defined by the presence of one or two copies of the E-ϕ-G-D-[KR]-[TS] consensus motif in their transmembrane domain. We previously showed that two members of this family, the human TMEM165 and the budding yeast Gdt1p, are functionally related and are likely to form a new group of Ca(2+) transporters. Mutations in TMEM165 have been demonstrated to cause a new type of rare human genetic diseases denominated as Congenital Disorders of Glycosylation...
January 23, 2017: Cellular Microbiology
https://www.readbyqxmd.com/read/28109583/detection-of-recombinant-human-lactoferrin-and-lysozyme-produced-in-a-bitransgenic-cow
#4
Germán G Kaiser, Nicolás C Mucci, Vega González, Lourdes Sánchez, José A Parrón, María D Pérez, Miguel Calvo, Juan F Aller, Federico A Hozbor, Adrián A Mutto
Lactoferrin and lysozyme are 2 glycoproteins with great antimicrobial activity, being part of the nonspecific defensive system of human milk, though their use in commercial products is difficult because human milk is a limited source. Therefore, many investigations have been carried out to produce those proteins in biological systems, such as bacteria, yeasts, or plants. Mammals seem to be more suitable as expression systems for human proteins, however, especially for those that are glycosylated. In the present study, we developed a bicistronic commercial vector containing a goat β-casein promoter and an internal ribosome entry site fragment between the human lactoferrin and human lysozyme genes to allow the introduction of both genes into bovine adult fibroblasts in a single transfection...
March 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/28089756/glycan-alteration-imparts-cellular-resistance-to-a-membrane-lytic-anticancer-peptide
#5
Ken Ishikawa, Scott H Medina, Joel P Schneider, Amar J S Klar
Although resistance toward small-molecule chemotherapeutics has been well studied, the potential of tumor cells to avoid destruction by membrane-lytic compounds remains unexplored. Anticancer peptides (ACPs) are a class of such agents that disrupt tumor cell membranes through rapid and non-stereospecific mechanisms, encouraging the perception that cellular resistance toward ACPs is unlikely to occur. We demonstrate that eukaryotic cells can, indeed, develop resistance to the model oncolytic peptide SVS-1, which preferentially disrupts the membranes of cancer cells...
January 9, 2017: Cell Chemical Biology
https://www.readbyqxmd.com/read/28087398/interaction-of-mal-de-r%C3%A3-o-cuarto-virus-fijivirus-genus-proteins-and-identification-of-putative-factors-determining-viroplasm-formation-and-decay
#6
Gabriela Llauger, Luis Alejandro de Haro, Victoria Alfonso, Mariana Del Vas
Mal de Río Cuarto virus (MRCV) is a member of the Fijivirus genus, within the Reoviridae family, that replicates and assembles in cytoplasmic inclusion bodies called viroplasms. In this study, we investigated interactions between ten MRCV proteins by yeast two-hybrid (Y2H) assays and identified interactions of non-structural proteins P6/P6, P9-2/P9-2 and P6/P9-1. P9-1 and P6 are the major and minor components of the viroplasms respectively, whereas P9-2 is an N-glycosylated membrane protein of unknown function...
January 10, 2017: Virus Research
https://www.readbyqxmd.com/read/28036287/n-glycosylation-converts-non-glycoproteins-into-mannose-receptor-ligands-and-reveals-antigen-specific-t-cell-responses-in-vivo
#7
Christoph Kreer, Janina M Kuepper, Matthias Zehner, Thomas Quast, Waldemar Kolanus, Beatrix Schumak, Sven Burgdorf
N-glycosylation is generally accepted to enhance the immunogenicity of antigens because of two main reasons. First, the attachment of glycans enables recognition by endocytic receptors like the mannose receptor (MR) and hence increased uptake by dendritic cells (DCs). Second, foreign glycans are postulated to be immunostimulatory and their recognition could induce DC activation. However, a direct comparison between the immunogenicity of N-glycosylated vs. de-glycosylated proteins in vivo and a direct effect of N-glycosylated antigens on the intrinsic capacity of DCs to activate T cells have not been assessed so far...
January 24, 2017: Oncotarget
https://www.readbyqxmd.com/read/28013188/a-capture-method-based-on-the-vc1-domain-reveals-new-binding-properties-of-the-human-receptor-for-advanced-glycation-end-products-rage
#8
Genny Degani, Alessandra A Altomare, Mara Colzani, Caterina Martino, Angelica Mazzolari, Guenter Fritz, Giulio Vistoli, Laura Popolo, Giancarlo Aldini
The Advanced Glycation and Lipoxidation End products (AGEs and ALEs) are a heterogeneous class of compounds derived from the non-enzymatic glycation or protein adduction by lipoxidation break-down products. The receptor for AGEs (RAGE) is involved in the progression of chronic diseases based on persistent inflammatory state and oxidative stress. RAGE is a pattern recognition receptor (PRR) and the inhibition of the interaction with its ligands or of the ligand accumulation have a potential therapeutic effect...
December 18, 2016: Redox Biology
https://www.readbyqxmd.com/read/28010776/rational-design-for-the-stability-improvement-of-armillariella-tabescens-%C3%AE-mannanase-man47-based-on-n-glycosylation-modification
#9
Weixiong Hu, Xiaoyun Liu, Yufeng Li, Daling Liu, Zhihe Kuang, Chuiwen Qian, Dongsheng Yao
β-Mannanase has been widely used in industries such as food and feed processing and thus has been a target enzyme for biotechnological development. In this study, we sought to improve the stability and protease resistance of a recombinant β-mannanase, MAN47 from Armillariella tabescens, through rationally designed N-glycosylation. Based on homology modeling, molecular docking, secondary structure analysis and glycosylation feasibility analysis, an enhanced aromatic sequon sequence was introduced into specific MAN47 loop regions to facilitate N-glycosylation...
February 2017: Enzyme and Microbial Technology
https://www.readbyqxmd.com/read/27998791/molecular-modelling-dynamics-simulation-and-characterization-of-antifungal-chitinase-from-sechium-edule
#10
Bipasha Bhattacharjee, Neeta Pathaw, Nikhil K Chrungoo, Atanu Bhattacharjee
Chitinases are varied sized proteins which have the ability to degrade chitin and are present in a huge range of organisms like fungi, yeasts, arthropods, humans etc. and have been getting increased attention due to their biocontrol properties. In silico analysis sheds light on the extensive properties of this plant protein. In this paper, a particular antifungal protein Chitinase sourced from Sechium edule from East Khasi Hills, Meghalaya was characterized using an array of bioinformatics tools. The modelled protein showed conserved domains characteristic to glycosyl hydrolase, family 18 superfamily...
March 30, 2017: Gene
https://www.readbyqxmd.com/read/27975137/expression-and-secretion-of-glycosylated-heparin-biosynthetic-enzymes-using-komagataella-pastoris
#11
Jacob A Englaender, Yuanyuan Zhu, Abhijit N Shirke, Lei Lin, Xinyue Liu, Fuming Zhang, Richard A Gross, Mattheos A G Koffas, Robert J Linhardt
Heparin, an anticoagulant drug, is biosynthesized in selected animal cells. The heparin biosynthetic enzymes mainly consist of sulfotransferases and all are integral transmembrane glycoproteins. These enzymes are generally produced in engineered Escherichia coli as without their transmembrane domains as non-glycosylated fusion proteins. In this study, we used the yeast, Komagataella pastoris, to prepare four sulfotransferases involved in heparin biosynthesis as glycoproteins. While the yields of these yeast-expressed enzymes were considerably lower than E...
December 14, 2016: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/27933904/global-analysis-of-secreted-proteins-and-glycoproteins-in-saccharomyces-cerevisiae
#12
Johanna M Smeekens, Haopeng Xiao, Ronghu Wu
Protein secretion is essential for numerous cellular activities, and secreted proteins in bodily fluids are a promising and noninvasive source of biomarkers for disease detection. Systematic analysis of secreted proteins and glycoproteins will provide insight into protein function and cellular activities. Yeast (Saccharomyces cerevisiae) is an excellent model system for eukaryotic cells, but global analysis of secreted proteins and glycoproteins in yeast is challenging due to the low abundances of secreted proteins and contamination from high-abundance intracellular proteins...
December 20, 2016: Journal of Proteome Research
https://www.readbyqxmd.com/read/27927990/rft1-protein-affects-glycosylphosphatidylinositol-gpi-anchor-glycosylation
#13
Petra Gottier, Amaia Gonzalez-Salgado, Anant K Menon, Yuk-Chien Liu, Alvaro Acosta-Serrano, Peter Bütikofer
The membrane protein RFT1 is essential for normal protein N-glycosylation, but its precise function is not known. RFT1 was originally proposed to translocate the glycolipid Man5GlcNAc2-PP-dolichol (needed to synthesize N-glycan precursors) across the endoplasmic reticulum membrane, but subsequent studies showed that it does not play a direct role in transport. In contrast to the situation in yeast, RFT1 is not essential for growth of the parasitic protozoan Trypanosoma brucei, enabling the study of its function in a null background...
January 20, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27921082/global-role-of-cyclic-amp-signaling-in-ph-dependent-responses-in-candida-albicans
#14
Jeffrey M Hollomon, Nora Grahl, Sven D Willger, Katja Koeppen, Deborah A Hogan
Candida albicans behaviors are affected by pH, an important environmental variable. Filamentous growth is a pH-responsive behavior, where alkaline conditions favor hyphal growth and acid conditions favor growth as yeast. We employed filamentous growth as a tool to study the impact of pH on the hyphal growth regulator Cyr1, and we report that downregulation of cyclic AMP (cAMP) signaling by acidic pH contributes to the inhibition of hyphal growth in minimal medium with GlcNAc. Ras1 and Cyr1 are generally required for efficient hyphal growth, and the effects of low pH on Ras1 proteolysis and GTP binding are consistent with diminished cAMP output...
November 2016: MSphere
https://www.readbyqxmd.com/read/27906032/infection-of-rhesus-macaques-with-a-pool-of-simian-immunodeficiency-virus-with-the-envelope-genes-from-acute-hiv-1-infections
#15
Kendall C Krebs, Meijuan Tian, Mohammed Asmal, Binhua Ling, Kenneth Nelson, Kenneth Henry, Richard Gibson, Yuejin Li, Weining Han, Robin J Shattock, Ronald S Veazey, Norman Letvin, Eric J Arts, Yong Gao
BACKGROUND: New simian-human immunodeficiency chimeric viruses with an HIV-1 env (SHIVenv) are critical for studies on HIV pathogenesis, vaccine development, and microbicide testing. Macaques are typically exposed to single CCR5-using SHIVenv which in most instances does not reflect the conditions during acute/early HIV infection (AHI) in humans. Instead of individual and serial testing new SHIV constructs, a pool of SHIVenv_B derived from 16 acute HIV-1 infections were constructed using a novel yeast-based SHIV cloning approach and then used to infect macaques...
November 25, 2016: AIDS Research and Therapy
https://www.readbyqxmd.com/read/27837408/humanizing-glycosylation-pathways-in-eukaryotic-expression-systems
#16
REVIEW
Amjad Hayat Khan, Hadi Bayat, Masoumeh Rajabibazl, Suriana Sabri, Azam Rahimpour
Glycosylation represents the most widespread posttranslational modifications, found in a broad spectrum of natural and therapeutic recombinant proteins. It highly affects bioactivity, site-specificity, stability, solubility, immunogenicity, and serum half-life of glycoproteins. Numerous expression hosts including yeasts, insect cells, transgenic plants, and mammalian cells have been explored for synthesizing therapeutic glycoproteins. However, glycosylation profile of eukaryotic expression systems differs from human...
January 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/27836640/transmembrane-topology-of-the-arsenite-permease-acr3-from-saccharomyces-cerevisiae
#17
Donata Wawrzycka, Katarzyna Markowska, Ewa Maciaszczyk-Dziubinska, Magdalena Migocka, Robert Wysocki
Acr3 is a plasma membrane transporter, a member of the bile/arsenite/riboflavin transporter (BART) superfamily, which confers high-level resistance to arsenicals in the yeast Saccharomyces cerevisiae. We have previously shown that the yeast Acr3 acts as a low affinity As(III)/H(+) and Sb(III)/H(+) antiporter. We have also identified several amino acid residues that are localized in putative transmembrane helices (TM) and appeared to be critical for the Acr3 activity. In the present study, the topology of Acr3 was investigated by insertion of glycosylation and factor Xa protease cleavage sites at predicted hydrophilic regions...
January 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27794496/review-of-the-recombinant-human-interferon-gamma-as-an-immunotherapeutic-impacts-of-production-platforms-and-glycosylation
#18
REVIEW
Ali Razaghi, Leigh Owens, Kirsten Heimann
Human interferon gamma is a cytokine belonging to a diverse group of interferons which have a crucial immunological function against mycobacteria and a wide variety of viral infections. To date, it has been approved for treatment of chronic granulomatous disease and malignant osteopetrosis, and its application as an immunotherapeutic agent against cancer is an increasing prospect. Recombinant human interferon gamma, as a lucrative biopharmaceutical, has been engineered in different expression systems including prokaryotic, protozoan, fungal (yeasts), plant, insect and mammalian cells...
December 20, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/27769287/effects-of-rho1-a-small-gtpase-on-the-production-of-recombinant-glycoproteins-in-saccharomyces-cerevisiae
#19
Sha Xu, Ge-Yuan Zhang, Huijie Zhang, Toshihiko Kitajima, Hideki Nakanishi, Xiao-Dong Gao
BACKGROUND: To humanize yeast N-glycosylation pathways, genes involved in yeast specific hyper-mannosylation must be disrupted followed by the introduction of genes catalyzing the synthesis, transport, and addition of human sugars. However, deletion of these genes, for instance, OCH1, which initiates hyper-mannosylation, could cause severe defects in cell growth, morphogenesis and response to environmental challenges. RESULTS: In this study, overexpression of RHO1, which encodes the Rho1p small GTPase, is confirmed to partially recover the growth defect of Saccharomyces cerevisiae Δalg3Δoch1 double mutant strain...
October 21, 2016: Microbial Cell Factories
https://www.readbyqxmd.com/read/27768707/dom34-links-translation-to-protein-o-mannosylation
#20
Lasse van Wijlick, René Geissen, Jessica S Hilbig, Quentin Lagadec, Pilar D Cantero, Eugen Pfeifer, Mateusz Juchimiuk, Sven Kluge, Stephan Wickert, Paula Alepuz, Joachim F Ernst
In eukaryotes, Dom34 upregulates translation by securing levels of activatable ribosomal subunits. We found that in the yeast Saccharomyces cerevisiae and the human fungal pathogen Candida albicans, Dom34 interacts genetically with Pmt1, a major isoform of protein O-mannosyltransferase. In C. albicans, lack of Dom34 exacerbated defective phenotypes of pmt1 mutants, while they were ameliorated by Dom34 overproduction that enhanced Pmt1 protein but not PMT1 transcript levels. Translational effects of Dom34 required the 5'-UTR of the PMT1 transcript, which bound recombinant Dom34 directly at a CA/AC-rich sequence and regulated in vitro translation...
October 2016: PLoS Genetics
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