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Xylosyltransferase 2

Sara C Sukenik, Kalimuthu Karuppanan, Qiongyu Li, Carlito B Lebrilla, Somen Nandi, Karen A McDonald
Transient recombinant protein production is a promising alternative to stable transgenic systems, particularly for emergency situations in which rapid production of novel therapeutics is needed. In plants, Agrobacterium tumefaciens can be used as a gene delivery vector for transient expression. A potential barrier for plant-based production of human therapeutics is that different glycosylation patterns are found on plant and mammalian proteins. Since glycosylation can affect the efficacy, safety and stability of a therapeutic protein, methods to control glycan structures and distributions in plant-based systems would be beneficial...
April 16, 2018: International Journal of Molecular Sciences
Kouki Matsuo, Go Atsumi
Through the years, the post-translational modification of plant-made recombinant proteins has been a considerable problem. Protein glycosylation is arguably the most important post-translational modification; thus, for the humanization of protein glycosylation in plants, the introduction, repression, and knockout of many glycosylation-related genes has been carried out. In addition, plants lack mammalian-type protein O-glycosylation pathways; thus, for the synthesis of mammalian O-glycans in plants, the construction of these pathways is necessary...
April 12, 2018: Journal of Bioscience and Bioengineering
Ruiqin Zhong, Dongtao Cui, Dennis R Phillips, Zheng-Hua Ye
Xylan is a major hemicellulose in both primary and secondary walls of grass species. It consists of a linear backbone of β-1,4-linked xylosyl residues that are often substituted with monosaccharides and disaccharides. Xylosyl substitutions directly on the xylan backbone have not been reported in grass species, and genes responsible for xylan substitutions in grass species have not been well elucidated. Here, we report functional characterization of a rice (Oryza sativa) GT61 glycosyltransferase, XYXT1 (xylan xylosyltransferase1), for its role in xylan substitutions...
March 1, 2018: Plant & Cell Physiology
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
Wei Zeng, Kristina L Ford, Antony Bacic, Joshua L Heazlewood
N -glycosylation is one of the most common protein post-translational modifications in eukaryotes and has a relatively conserved core structure between fungi, animals and plants. In plants, the biosynthesis of N -glycans has been extensively studied with all the major biosynthetic enzymes characterized. However, few studies have applied advanced mass spectrometry to profile intact plant N -glycopeptides. In this study, we use hydrophilic enrichment, high-resolution tandem mass spectrometry with complementary and triggered fragmentation to profile Arabidopsis N -glycopeptides from microsomal membranes of aerial tissues...
March 2018: Molecular & Cellular Proteomics: MCP
Pietro Dallabernardina, Colin Ruprecht, Peter J Smith, Michael G Hahn, Breeanna R Urbanowicz, Fabian Pfrengle
We report the automated glycan assembly of oligosaccharides related to the plant cell wall hemicellulosic polysaccharide xyloglucan. The synthesis of galactosylated xyloglucan oligosaccharides was enabled by introducing p-methoxybenzyl (PMB) as a temporary protecting group for automated glycan assembly. The generated oligosaccharides were printed as microarrays, and the binding of a collection of xyloglucan-directed monoclonal antibodies (mAbs) to the oligosaccharides was assessed. We also demonstrated that the printed glycans can be further enzymatically modified while appended to the microarray surface by Arabidopsis thaliana xyloglucan xylosyltransferase 2 (AtXXT2)...
December 6, 2017: Organic & Biomolecular Chemistry
M Umair, G Eckstein, G Rudolph, T Strom, E Graf, D Hendig, J Hoover, J Alanay, T Meitinger, H Schmidt, W Ahmad
Spondyloocular syndrome (SOS) is a rare autosomal recessive, skeletal disorder. Two recent studies have shown that it is the result of biallelic sequence variants in the XYLT2 gene with pleiotropic effects in multiple organs, including retina, heart muscle, inner ear, cartilage, and bone. The XYLT2 gene encodes xylosyltransferase 2, which catalyzes the transfer of xylose (monosaccharide) to the core protein of proteoglycans (PGs) leading to initiating the process of PG assembly. SOS was originally characterized in 2 families A and B of Iraqi and Turkish origin, respectively...
April 2018: Clinical Genetics
Can Liu, Baomei Wang, Zhaoxia Li, Zhenghua Peng, Juren Zhang
NAC proteins constitute one of the largest families of plant-specific transcription factors, and a number of these proteins participate in the regulation of plant development and responses to abiotic stress. T. HALOPHILA STRESS RELATED NAC1 ( TsNAC1 ), cloned from the halophyte Thellungiella halophila , is a NAC transcription factor gene, and its overexpression can improve abiotic stress resistance, especially in salt stress tolerance, in both T. halophila and Arabidopsis ( Arabidopsis thaliana ) and retard the growth of these plants...
January 2018: Plant Physiology
Grit Sandig, Hans Henning von Horsten, Lars Radke, Véronique Blanchard, Marcus Frohme, Christoph Giese, Volker Sandig, Stephan Hinderlich
Xylose is a general component of O-glycans in mammals. Core-xylosylation of N-glycans is only found in plants and helminth. Consequently, xylosylated N-glycans cause immunological response in humans. We have used the F-protein of the human respiratory syncytial virus (RSV), one of the main causes of respiratory tract infection in infants and elderly, as a model protein for vaccination. The RSV-F protein was expressed in CHO-DG44 cells, which were further modified by co-expression of β1,2-xylosyltransferase from Nicotiana tabacum...
April 28, 2017: Bioengineering
Sébastien Mercx, Nicolas Smargiasso, François Chaumont, Edwin De Pauw, Marc Boutry, Catherine Navarre
Plants or plant cells can be used to produce pharmacological glycoproteins such as antibodies or vaccines. However these proteins carry N-glycans with plant-typical residues [β(1,2)-xylose and core α(1,3)-fucose], which can greatly impact the immunogenicity, allergenicity, or activity of the protein. Two enzymes are responsible for the addition of plant-specific glycans: β(1,2)-xylosyltransferase (XylT) and α(1,3)-fucosyltransferase (FucT). Our aim consisted of knocking-out two XylT genes and four FucT genes (12 alleles altogether) in Nicotiana tabacum BY-2 suspension cells using CRISPR/Cas9...
2017: Frontiers in Plant Science
T V Komarova, E V Sheshukova, E N Kosobokova, M V Serebryakova, V S Kosorukov, V N Tashlitsky, Y L Dorokhov
Plant biosimilars of anticancer therapeutic antibodies are of interest not only because of the prospects of their practical use, but also as an instrument and object for study of plant protein glycosylation. In this work, we first designed a pertuzumab plant biosimilar (PPB) and investigated the composition of its Asn297-linked glycan in comparison with trastuzumab plant biosimilar (TPB). Both biosimilars were produced in wild-type (WT) Nicotiana benthamiana plant (PPB-WT and TPB-WT) and transgenic ΔXTFT N...
April 2017: Biochemistry. Biokhimii︠a︡
Boyan Grigorov, Emma Reungoat, Alice Gentil Dit Maurin, Mihayl Varbanov, Julie Blaising, Maud Michelet, Rachel Manuel, Romain Parent, Birke Bartosch, Fabien Zoulim, Florence Ruggiero, Eve-Isabelle Pécheur
The hepatitis C virus (HCV) infects hepatocytes after binding to heparan sulfate proteoglycans, in particular Syndecan-1, followed by recognition of the tetraspanin CD81 and other receptors. Heparan sulfate proteoglycans are found in a specific microenvironment coating the hepatocyte surface called the glycocalyx and are receptors for extracellular matrix proteins, cytokines, growth factors, lipoproteins, and infectious agents. We investigated the mutual influence of HCV infection on the glycocalyx and revealed new links between Syndecan-1 and CD81...
May 2017: Cellular Microbiology
Long Guo, Nursel H Elcioglu, Aritoshi Iida, Yasemin K Demirkol, Seda Aras, Naomichi Matsumoto, Gen Nishimura, Noriko Miyake, Shiro Ikegawa
Desbuquois dysplasia (DBQD) is an autosomal recessive skeletal disorder characterized by growth retardation, joint laxity, short extremities, and progressive scoliosis. DBQD is classified into two types based on the presence (DBQD1) or absence (DBQD2) of characteristic hand abnormalities. CANT1 mutations have been reported in both DBQD1 and DBQD2. Recently, mutations in the gene encoding xylosyltransferase 1 (XYLT1) were identified in several families with DBQD2. In this study, we performed whole-exome sequencing in two Turkish families with DBQD2...
March 2017: Journal of Human Genetics
Lu Yao, Li-Ping Zhu, Xiao-Yan Xu, Ling-Ling Tan, Martin Sadilek, Huan Fan, Bo Hu, Xiao-Ting Shen, Jie Yang, Bin Qiao, Song Yang
Transcriptomic analysis of cultured fungi suggests that many genes for secondary metabolite synthesis are presumably silent under standard laboratory condition. In order to investigate the expression of silent genes in symbiotic systems, 136 fungi-fungi symbiotic systems were built up by co-culturing seventeen basidiomycetes, among which the co-culture of Trametes versicolor and Ganoderma applanatum demonstrated the strongest coloration of confrontation zones. Metabolomics study of this co-culture discovered that sixty-two features were either newly synthesized or highly produced in the co-culture compared with individual cultures...
2016: Scientific Reports
Alan T Culbertson, Adrienne L Smith, Matthew D Cook, Olga A Zabotina
Xyloglucan is the most abundant hemicellulose in the primary cell wall of dicotyledonous plants. In Arabidopsis, three xyloglucan xylosyltransferases, XXT1, XXT2, and XXT5, participate in xylosylation of the xyloglucan backbone. Despite the importance of these enzymes, there is a lack of information on their structure and the critical residues required for substrate binding and transferase activity. In this study, the roles of different domains of XX2 in protein expression and catalytic activity were investigated by constructing a series of N- and C-terminal truncations...
August 2016: Phytochemistry
Alan T Culbertson, Alesia A Tietze, Daniel Tietze, Yi-Hsiang Chou, Adrienne L Smith, Zachary T Young, Olga A Zabotina
In dicotyledonous plants, xyloglucan (XyG) is the most abundant hemicellulose of the primary cell wall. The enzymes involved in XyG biosynthesis have been identified through reverse-genetics and activity was characterized by heterologous expression. Currently, there is no information on the atomic structures or amino acids involved in activity or substrate binding of any of the Golgi-localized XyG biosynthetic enzymes. A homology model of the xyloglucan xylosyltransferase 2 (XXT2) catalytic domain was built on the basis of the crystal structure of A64Rp...
September 2016: Glycobiology
Aleksander Jamsheer, Ewelina M Olech, Kazimierz Kozłowski, Marek Niedziela, Anna Sowińska-Seidler, Monika Obara-Moszyńska, Anna Latos-Bieleńska, Marek Karczewski, Tomasz Zemojtel
Desbuquois dysplasia type 2 (DBQD2) is a rare recessively inherited skeletal genetic disorder characterized by severe prenatal and postnatal growth retardation, generalized joint laxity with dislocation of large joints and facial dysmorphism. The condition was recently described to result from autosomal recessive mutations in XYLT1, encoding the enzyme xylosyltransferase-1. In this paper, we report on a Polish patient with DBQD2 who presented with severe short stature of prenatal onset, joint laxity, psychomotor retardation and multiple radiological abnormalities including short metacarpals, advanced bone age and exaggerated trochanters...
July 2016: Journal of Human Genetics
Craig F Munns, Somayyeh Fahiminiya, Nabin Poudel, Maria Cristina Munteanu, Jacek Majewski, David O Sillence, Jordan P Metcalf, Andrew Biggin, Francis Glorieux, François Fassier, Frank Rauch, Myron E Hinsdale
Heparan and chondroitin/dermatan sulfated proteoglycans have a wide range of roles in cellular and tissue homeostasis including growth factor function, morphogen gradient formation, and co-receptor activity. Proteoglycan assembly initiates with a xylose monosaccharide covalently attached by either xylosyltransferase I or II. Three individuals from two families were found that exhibited similar phenotypes. The index case subjects were two brothers, individuals 1 and 2, who presented with osteoporosis, cataracts, sensorineural hearing loss, and mild learning defects...
June 4, 2015: American Journal of Human Genetics
Sho Takano, Shuichi Matsuda, Atsushi Funabiki, Jun-ichi Furukawa, Takaki Yamauchi, Yoshihiko Tokuji, Mikio Nakazono, Yasuro Shinohara, Itsuro Takamure, Kiyoaki Kato
Seed germination rates and plant development and growth under abiotic stress are important aspects of crop productivity. Here, our characterization of the rice (Oryza sativa L.) mutant reduced culm number11 (rcn11) showed that RCN11 controls growth of plants exposed to abnormal temperature, salinity and drought conditions. RCN11 also mediates root aerenchyma formation under oxygen-deficient conditions and ABA sensitivity during seed germination. Molecular studies showed that the rcn11 mutation resulted from a 966-bp deletion that caused loss of function of β1,2-xylosyltransferase (OsXylT)...
July 2015: Plant Science: An International Journal of Experimental Plant Biology
Jin Li, Thomas J Stoddard, Zachary L Demorest, Pierre-Olivier Lavoie, Song Luo, Benjamin M Clasen, Frederic Cedrone, Erin E Ray, Andrew P Coffman, Aurelie Daulhac, Ann Yabandith, Adam J Retterath, Luc Mathis, Daniel F Voytas, Marc-André D'Aoust, Feng Zhang
Biopharmaceutical glycoproteins produced in plants carry N-glycans with plant-specific residues core α(1,3)-fucose and β(1,2)-xylose, which can significantly impact the activity, stability and immunogenicity of biopharmaceuticals. In this study, we have employed sequence-specific transcription activator-like effector nucleases (TALENs) to knock out two α(1,3)-fucosyltransferase (FucT) and the two β(1,2)-xylosyltransferase (XylT) genes within Nicotiana benthamiana to generate plants with improved capacity to produce glycoproteins devoid of plant-specific residues...
February 2016: Plant Biotechnology Journal
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