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Glycosyltransferase

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https://www.readbyqxmd.com/read/29156217/characterization-of-udp-glucosyltransferase-from-indigofera-tinctoria
#1
Shintaro Inoue, Toshiki Moriya, Rihito Morita, Keiko Kuwata, Sanjog T Thul, Bijaya K Sarangi, Yoshiko Minami
Indican is a secondary metabolite in Indigofera tinctoria; its synthesis from indoxyl and UDP-glucose is catalyzed by a UDP-glucosyltransferase (UGT). In this study, we partially purified UGT extracted from I. tinctoria leaves and analyzed the protein by peptide mass fingerprinting. We identified two fragments that were homologous to UGT after comparison with the transcriptomic data of I. tinctoria leaves. The fragments were named itUgt1 and itUgt2 and were amplified using rapid amplification of cDNA ends polymerase chain reaction to obtain full-length cDNAs...
November 6, 2017: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/29156140/automated-training-of-reaxff-reactive-force-fields-for-energetics-of-enzymatic-reactions
#2
Tomáš Trnka, Igor Tvaroska, Jaroslav Koca
Computational studies of the reaction mechanisms of various enzymes are nowadays based almost exclusively on hybrid QM/MM models. Unfortunately, the success of this approach strongly depends on the selection of the QM region, and computational cost is a crucial limiting factor. An interesting alternative is offered by empirical reactive molecular force fields, especially the ReaxFF potential developed by van Duin and co-workers. However, even though an initial parameterization of ReaxFF for biomolecules already exists, it does not provide the desired level of accuracy...
November 20, 2017: Journal of Chemical Theory and Computation
https://www.readbyqxmd.com/read/29143814/structural-basis-of-nucleotide-sugar-transport-across-the-golgi-membrane
#3
Joanne L Parker, Simon Newstead
Glycosylation is a fundamental cellular process that, in eukaryotes, occurs in the lumen of both the Golgi apparatus and the endoplasmic reticulum. Nucleotide sugar transporters (NSTs) are an essential component of the glycosylation pathway, providing the diverse range of substrates required for the glycosyltransferases. NSTs are linked to several developmental and immune disorders in humans, and in pathogenic microbes they have an important role in virulence. How NSTs recognize and transport activated monosaccharides, however, is currently unclear...
November 15, 2017: Nature
https://www.readbyqxmd.com/read/29140092/determinants-and-expansion-of-specificity-in-a-trichothecene-udp-glucosyltransferase-from-oryza-sativa
#4
Karl M Wetterhorn, Kaitlyn Gabardi, Herbert Michlmayr, Alexandra Malachova, Mark Busman, Susan Patricia McCormick, Franz Berthiller, Gerhard Adam, Ivan Rayment
Family 1 UDP-glycosyltransferases in plants (UGTs) primarily form glucose conjugates of small molecules and, besides other functions, play a role in detoxification of xenobiotics. Indeed, overexpression of a barley UGT in wheat has been shown to control Fusarium head blight, where this is a plant disease of global significance that leads to reduced crop yields and contamination with trichothecene mycotoxins such as deoxynivalenol (DON), T-2 toxin and many other structural variants. The UGT Os79 from rice has emerged as a promising candidate for inactivation of mycotoxins on account of its ability to glycosylate DON, nivalenol and hydrolyzed T-2 toxin (HT-2)...
November 15, 2017: Biochemistry
https://www.readbyqxmd.com/read/29140084/characterization-of-a-ugt84-family-glycosyltransferase-provides-new-insights-into-substrate-binding-and-reactivity-of-galloylglucose-ester-forming-ugts
#5
Alexander E Wilson, Xiaoxue Feng, Nadia N Ono, Doron Holland, Rachel Amir, Li Tian
Galloylated plant specialized metabolites play important roles in plant-environment interactions as well as the promotion of human and animal health. The galloylation reactions are mediated by the formation of galloylglucose esters from gallic acid and UDP-glucose, catalyzed by the plant UGT84 family glycosyltransferases. To explore and exploit the structural determinants of UGT84 activities, we performed homology modeling and substrate docking of PgUGT84A23, a galloylglucose ester-forming family 84 UGT, as well as sequence comparisons of PgUGT84A23 with other functionally characterized plant UGTs...
November 15, 2017: Biochemistry
https://www.readbyqxmd.com/read/29138972/stable-isotope-labeling-and-2-3-5-4-tetrahydroxystilbene-2-o-%C3%AE-d-glucopyranoside-biosynthetic-pathway-characterization-in-fallopia-multiflora
#6
Wanxia Xia, Wen Rui, Wei Zhao, Shujing Sheng, Lei Lei, Yifan Feng, Shujin Zhao
The THSG biosynthetic pathway in F. multiflora was characterized, and enzymatic activities responsible for the resveratrol synthesis, hydroxylation, and glycosylation reactions involved in THSG biosynthesis were confirmed in vitro. The biosynthetic origin of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside (THSG) and the enzymes involved in THSG biosynthesis in Fallopia multiflora were studied using stable isotope labeling and biocatalytic methods. UPLC-MS-based analyses were used to unravel the isotopologue composition of the biosynthetic intermediates and products, as well as to detect the products of the enzyme assay experiments...
November 14, 2017: Planta
https://www.readbyqxmd.com/read/29131612/functional-characterization-of-epitheaflagallin-3-o-gallate-generated-in-laccase-treated-green-tea-extracts-in-the-presence-of-gallic-acid
#7
Nobuya Itoh, Junji Kurokawa, Yasuhiro Isogai, Masaru Ogasawara, Takayuki Matsunaga, Tutomu Okubo, Yuji Katsube
Epitheaflagallin (ETFG) and epitheaflagallin 3-O-gallate (ETFGg) are minor polyphenols in black tea extract that are enzymatically synthesized from epigallocatechin (EGC) and epigallocatechin gallate (EGCg), respectively, in green tea extract via laccase oxidation in the presence of gallic acid. The constituents of laccase-treated green tea extract in the presence of gallic acid are thus quite different from those of non-laccase-treated green tea extract: EGC and EGCg are present in lower concentrations, and ETFG and ETFGg are present in higher concentrations...
November 13, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/29131308/glycosyltransferase-cascades-made-fit-for-chemical-production-integrated-biocatalytic-process-for-the-natural-polyphenol-c-glucoside-nothofagin
#8
Katharina Schmölzer, Martin Lemmerer, Bernd Nidetzky
Glycosyltransferase cascades are promising tools of biocatalysis for natural product glycosylation, but their suitability for actual production remains to be shown. Here, we demonstrate at a scale of 100 g isolated product the integrated biocatalytic production of nothofagin, the natural 3'-C-β-D-glucoside of the polyphenol phloretin. A parallel reaction cascade involving coupled C-glucosyltransferase and sucrose synthase was optimized for the one-pot glucosylation of phloretin from sucrose via an UDP/UDP-glucose shuttle...
November 13, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/29127201/loss-of-the-mucosal-barrier-alters-the-progenitor-cell-niche-via-jak-stat-signaling
#9
Liping Zhang, Katharina Ribbeck, Bradley Turner, Kelly G Ten Hagen
The mucous barrier of our digestive tract is the first line of defense against pathogens and damage. Disruptions in this barrier are associated with diseases such as Crohn's disease, colitis and colon cancer, but mechanistic insights into these processes and diseases are limited. We have previously shown that loss of a conserved O-glycosyltransferase (PGANT4) in Drosophila results in aberrant secretion of components of the peritrophic/mucous membrane in the larval digestive tract. Here, we show that loss of pgant4 disrupts the mucosal barrier, resulting in epithelial expression of the IL-6-like cytokine Upd3, leading to activation of JAK/STAT signaling, differentiation of cells that form the progenitor cell niche and abnormal proliferation of progenitor cells...
November 10, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29120519/palovarotene-inhibits-osteochondroma-formation-in-a-mouse-model-of-multiple-hereditary-exostoses
#10
Toshihiro Inubushi, Isabelle Lemire, Fumitoshi Irie, Yu Yamaguchi
Multiple hereditary exostoses (MHE), also known as multiple osteochondromas (MO), is an autosomal dominant disorder characterized by the development of multiple cartilage-capped bone tumors (osteochondromas). The large majority of patients with MHE carry loss-of-function mutations in the EXT1 or EXT2 gene, which encodes a glycosyltransferase essential for heparan sulfate (HS) biosynthesis. Increasing evidence suggests that enhanced BMP signaling resulting from loss of HS expression plays a role in osteochondroma formation in MHE...
November 9, 2017: Journal of Bone and Mineral Research: the Official Journal of the American Society for Bone and Mineral Research
https://www.readbyqxmd.com/read/29115405/alterations-of-protein-glycosylation-in-embryonic-stem-cells-during-adipogenesis
#11
Wenguang Liu, Xingrong Yan, Wei Liu, Yangyang Wang, Yang Rao, Hanjie Yu, Jihong Cui, Xin Xie, Mei Sun, Lu Yin, Hongmin Li, Fulin Chen
The understanding of adipose tissue development is crucial for the treatment of obesity-related diseases. Adipogenesis has been extensively investigated at the gene and protein levels in recent years. However, the alterations in protein glycosylation during this process remains unknown, particularly that of parthenogenetic embryonic stem cells (pESCs), a type of ESCs with low immunogenicity and no ethical concerns regarding their use. Protein glycosylation markedly affects cell growth and development, cell-to-cell communication, tumour growth and metastasis...
November 7, 2017: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29113801/producing-a-glycosylating-escherichia-coli-cell-factory-the-placement-of-the-bacterial-oligosaccharyl-transferase-pglb-onto-the-genome
#12
Benjamin Strutton, Stephen R P Jaffé, Jagroop Pandhal, Phillip C Wright
Although Escherichia coli has been engineered to perform N-glycosylation of recombinant proteins, an optimal glycosylating strain has not been created. By inserting a codon optimised Campylobacter oligosaccharyltransferase onto the E. coli chromosome, we created a glycoprotein platform strain, where the target glycoprotein, sugar synthesis and glycosyltransferase enzymes, can be inserted using expression vectors to produce the desired homogenous glycoform. To assess the functionality and glycoprotein producing capacity of the chromosomally based OST, a combined Western blot and parallel reaction monitoring mass spectrometry approach was applied, with absolute quantification of glycoprotein...
November 4, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29109183/mutation-and-suppressor-analysis-of-the-essential-lps-transport-protein-lpta-reveals-strategies-to-overcome-severe-outer-membrane-permeability-defects-in-escherichia-coli
#13
Federica A Falchi, Elisa A Maccagni, Simone Puccio, Clelia Peano, Cristina De Castro, Angelo Palmigiano, Domenico Garozzo, Alessandra M Martorana, Alessandra Polissi, Gianni Dehò, Paola Sperandeo
In Gram-negative bacteria, lipopolysaccharide (LPS) contributes to the robust permeability barrier of the outer membrane (OM), preventing the entry of toxic molecules such as detergents and antibiotics. LPS is transported from the inner membrane (IM) to the OM by the Lpt multiprotein machinery. Defects in LPS transport compromise LPS assembly at the OM and result in increased antibiotic sensitivity. LptA is a key component of the Lpt machine that interacts with the IM protein LptC and chaperons LPS through the periplasm...
November 6, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/29106707/c-518t%C3%A2-%C3%A2-c-missense-mutation-in-the-b-glycosyltransferase-gene-responsible-for-a-weak-b-variant
#14
Yanling Ying, Xiaozhen Hong, Shu Chen, Ji He, Faming Zhu
No abstract text is available yet for this article.
November 6, 2017: Transfusion
https://www.readbyqxmd.com/read/29101090/a-biosynthetic-route-for-polysialylating-proteins-in-escherichia-coli
#15
Timothy G Keys, Michael Wetter, Ivan Hang, Christoph Rutschmann, Simona Russo, Manuela Mally, Michael Steffen, Matthias Zuppiger, Fabian Müller, Jörg Schneider, Amirreza Faridmoayer, Chia-Wei Lin, Markus Aebi
Polysialic acid (polySia) is a posttranslational modification found on only a handful of proteins in the central nervous and immune systems. The addition of polySia to therapeutic proteins improves pharmacokinetics and reduces immunogenicity. To date, polysialylation of therapeutic proteins has only been achieved in vitro by chemical or chemoenzymatic strategies. In this work, we develop a biosynthetic pathway for site-specific polysialylation of recombinant proteins in the cytoplasm of Escherichia coli. The pathway takes advantage of a bacterial cytoplasmic polypeptide-glycosyltransferase to establish a site-specific primer on the target protein...
November 1, 2017: Metabolic Engineering
https://www.readbyqxmd.com/read/29100177/acinetobacter-baumannii-k13-and-k73-capsular-polysaccharides-differ-only-in-k-unit-side-branches-of-novel-non-2-ulosonic-acids-di-n-acetylated-forms-of-either-acinetaminic-acid-or-8-epiacinetaminic-acid
#16
Johanna J Kenyon, Anastasiya A Kasimova, Anna Notaro, Nikolay P Arbatsky, Immacolata Speciale, Alexander S Shashkov, Cristina De Castro, Ruth M Hall, Yuriy A Knirel
Structures of capsular polysaccharides of Acinetobacter baumannii isolates carrying KL13 and KL73 gene clusters were established. The closely related KL73 and KL13 gene clusters differ only by one gene in the module responsible for synthesis of the non-2-ulosonic acids. The K13 and K73 polysaccharides differ only in a single side-chain sugar, which is either 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-altro- or -d-glycero-l-altro-non-2-ulosonic acid [di-N-acetylated forms of acinetaminic acid (Aci5Ac7Ac) or 8-epiacinetaminic acid (8eAci5Ac7Ac), respectively]...
November 27, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/29095157/pteridine-glycosyltransferase-from-chlorobium-tepidum-crystallization-and-x-ray-analysis
#17
Asaithambi Killivalavan, Young Shik Park, Kon Ho Lee
The pteridine glycosyltransferase (PGT) found in Chlorobium tepidum (CtPGT) catalyzes the conversion of L-threo-tetrahydrobiopterin to 1-O-(L-threo-biopterin-2'-yl)-β-N-acetylglucosamine using UDP-N-acetylglucosamine. The gene for CtPGT was cloned, and selenomethionine-derivatized protein was overexpressed and purified using various chromatographic techniques. The protein was crystallized by the hanging-drop vapour-diffusion method using 0.24 M triammonium citrate pH 7.0, 14%(w/v) PEG 3350 as a reservoir solution...
November 1, 2017: Acta Crystallographica. Section F, Structural Biology Communications
https://www.readbyqxmd.com/read/29093022/giantin-knockout-models-reveal-a-feedback-loop-between-golgi-function-and-glycosyltransferase-expression
#18
Nicola L Stevenson, Dylan J M Bergen, Roderick E H Skinner, Erika Kague, Elizabeth Martin-Silverstone, Kate A Robson Brown, Chrissy L Hammond, David J Stephens
The Golgi is the cellular hub for complex glycosylation, controlling accurate processing of complex proteoglycans, receptors, ligands, and glycolipids. Its structure and organisation is dependent on golgins, which tether cisternal membranes and incoming transport vesicles. Here we show that knockout of the largest golgin, giantin, leads to substantial changes in gene expression despite only limited effects on Golgi structure. Notably, 22 Golgi-resident glycosyltransferases, but not glycan processing enzymes or the ER glycosylation machinery, are differentially expressed following giantin ablation...
November 1, 2017: Journal of Cell Science
https://www.readbyqxmd.com/read/29080432/base-modified-gdp-mannose-derivatives-and-their-substrate-activity-towards-a-yeast-mannosyltransferase
#19
Alice Collier, Gerd K Wagner
We have previously developed a new class of inhibitors and chemical probes for glycosyltransferases through base-modification of the sugar-nucleotide donor. The key feature of these donor analogues is the presence of an additional substituent at the nucleobase. To date, the application of this general concept has been limited to UDP-sugars and UDP-sugar-dependent glycosyltransferases. Herein, we report for the first time the application of our approach to a GDP-mannose-dependent mannosyltransferase. We have prepared four GDP-mannose derivatives with an additional substituent at either position 6 or 8 of the nucleobase...
November 27, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/29077008/the-glycosyltransferases-of-lps-core-a-review-of-four-heptosyltransferase-enzymes-in-context
#20
REVIEW
Joy M Cote, Erika A Taylor
Bacterial antibiotic resistance is a rapidly expanding problem in the world today. Functionalization of the outer membrane of Gram-negative bacteria provides protection from extracellular antimicrobials, and serves as an innate resistance mechanism. Lipopolysaccharides (LPS) are a major cell-surface component of Gram-negative bacteria that contribute to protecting the bacterium from extracellular threats. LPS is biosynthesized by the sequential addition of sugar moieties by a number of glycosyltransferases (GTs)...
October 27, 2017: International Journal of Molecular Sciences
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