The DXD motif is required for GM2 synthase activity but is not critical for nucleotide binding.

Abstract:

:We tested the importance of the aspartate-any residue-aspartate (DXD) motif for the enzymatic activity and nucleotide binding capacity of the Golgi glycosyltransferase GM2 synthase. We prepared point mutations of the motif, which is found in the sequence 352-VLWVDDDFV, and analyzed cells that stably expressed the mutated proteins. Whereas the folding of the mutated proteins was not seriously disrupted as judged by assembly into homodimers, Golgi localization, and secretion of a soluble form of the enzyme, exchange of the highly conserved aspartic acid residues at position 356 or 358 with alanine or asparagine reduced enzyme activity to background levels. In contrast, the D356E and D357N mutations retained weak activity, while the activity of V352A and W354A mutants was 167% and 24% that of wild-type enzyme, respectively. Despite the major effect of the DXD motif on enzymatic activity, nucleotide binding was not altered in the triple mutant D356N/D357N/D358N as revealed by binding to UDP-beads and labeling with the photoaffinity reagent, P(3)-(4-azidoanilido)uridine 5'-triphosphate (AAUTP). In summary, rather than being critical for nucleotide binding, this motif may function during catalysis in GM2 synthase, as has been proposed elsewhere for the SpsA glycosyltransferase based on its crystal structure.

journal_name

Glycobiology

journal_title

Glycobiology

authors

Li J,Rancour DM,Allende ML,Worth CA,Darling DS,Gilbert JB,Menon AK,Young WW Jr

doi

10.1093/glycob/11.3.217

subject

Has Abstract

pub_date

2001-03-01 00:00:00

pages

217-29

issue

3

eissn

0959-6658

issn

1460-2423

journal_volume

11

pub_type

杂志文章
  • Preparation and structure of heparin lyase-derived heparan sulfate oligosaccharides.

    abstract::Porcine intestinal mucosal heparan sulfate was exhaustively depolymerized on a large scale using heparin lyase II (heparinase II) or heparin lyase III (heparitinase, EC 4.2.2.8). The oligosaccharide mixtures formed with each enzyme were fractionated by low pressure gel permeation chromatography. Size-uniform mixtures ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/7.2.231

    authors: Hileman RE,Smith AE,Toida T,Linhardt RJ

    更新日期:1997-03-01 00:00:00

  • Glycoprotein biosynthesis in yeast: purification and characterization of the endoplasmic reticulum Man9 processing alpha-mannosidase.

    abstract::Saccharomyces cerevisiae Man9-alpha-mannosidase, responsible for trimming Man9GlcNAc2 in the endoplasmic reticulum to Man8GlcNAc2, the substrate for oligosaccharide elongation, has been purified to homogeneity from stabilized microsomal membranes without employing autolytic digestion. The activity was solubilized by t...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/1.6.605

    authors: Ziegler FD,Trimble RB

    更新日期:1991-12-01 00:00:00

  • The effect of oral treatment with 6-O-butanoyl castanospermine (MDL 28,574) in the murine zosteriform model of HSV-1 infection.

    abstract::Oral treatment of mice, cutaneously infected with herpes simplex virus type 1 (HSV-1) (strain SC16), with the alpha-glucosidase 1 inhibitor 6-O-butanoyl castanospermine (MDL 28,574) produced a significant delay in lesion development and reduced the amount of virus recovered from the brain. Virus load in the brains of ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/5.2.249

    authors: Bridges CG,Ahmed SP,Kang MS,Nash RJ,Porter EA,Tyms AS

    更新日期:1995-03-01 00:00:00

  • Enzymically inactive members of the trans-sialidase family from Trypanosoma cruzi display beta-galactose binding activity.

    abstract::trans-sialidase is a unique sialidase in that, instead of hydrolizing sialic acid, it preferentially transfers the monosaccharide to a terminal beta-galactose in glycoproteins and glycolipids. This enzyme, originally identified in Trypanosoma cruzi, belongs to a large family of proteins. Some members of the family lac...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/9.6.581

    authors: Cremona ML,Campetella O,Sánchez DO,Frasch AC

    更新日期:1999-06-01 00:00:00

  • Effect of sypQ gene on poly-N-acetylglucosamine biosynthesis in Vibrio parahaemolyticus and its role in infection process.

    abstract::The syp locus includes four genes encoding putative regulators, six genes encoding glycosyltransferases, two encoding export proteins, and six other genes encoding unidentified functional proteins associated with biofilm formation and symbiotic colonization. However, the individual functions of the respective genes re...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwu001

    authors: Ye L,Zheng X,Zheng H

    更新日期:2014-04-01 00:00:00

  • Incorporation of 15N from ammonium into the N-linked oligosaccharides of an immunoadhesin glycoprotein expressed in Chinese hamster ovary cells.

    abstract::Elevated ammonium concentrations in the medium of cultivated cells have been shown to increase the intracellular levels of uridine-5'-diphospho-N-acetylglucosamine (UDP-GlcNAc) and uridine-5'-diphospho-N-acetylgalactosamine (UDP-GalNAc; Ryll et al., 1994). These sugar nucleotides are substrates for glycosyltransferase...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/9.2.125

    authors: Gawlitzek M,Papac DI,Sliwkowski MB,Ryll T

    更新日期:1999-02-01 00:00:00

  • Different acceptor specificities of two glucuronyltransferases involved in the biosynthesis of HNK-1 carbohydrate.

    abstract::The biosynthesis of HNK-1 carbohydrate is mainly regulated by two glucuronyltransferases (GlcAT-P and GlcAT-S) and a sulfotransferase (HNK-1 ST). To determine how the two glucuronyltransferases are involved in the biosynthesis of the HNK-1 carbohydrate, we prepared soluble forms of GlcAT-P and GlcAT-S fused with the I...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwi001

    authors: Kakuda S,Sato Y,Tonoyama Y,Oka S,Kawasaki T

    更新日期:2005-02-01 00:00:00

  • Lactose binding to human galectin-7 (p53-induced gene 1) induces long-range effects through the protein resulting in increased dimer stability and evidence for positive cooperativity.

    abstract::The product of p53-induced gene 1 is a member of the galectin family, i.e., galectin-7 (Gal-7). To move beyond structural data by X-ray diffraction, we initiated the study of the lectin by nuclear magnetic resonance (NMR) and circular dichroism spectroscopies, and molecular dynamics (MD) simulations. In concert, our r...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwt005

    authors: Ermakova E,Miller MC,Nesmelova IV,López-Merino L,Berbís MA,Nesmelov Y,Tkachev YV,Lagartera L,Daragan VA,André S,Cañada FJ,Jiménez-Barbero J,Solís D,Gabius HJ,Mayo KH

    更新日期:2013-05-01 00:00:00

  • A novel antibody for human induced pluripotent stem cells and embryonic stem cells recognizes a type of keratan sulfate lacking oversulfated structures.

    abstract::We have generated a monoclonal antibody (R-10G) specific to human induced pluripotent stem (hiPS)/embryonic stem (hES) cells by using hiPS cells (Tic) as an antigen, followed by differential screening of mouse hybridomas with hiPS and human embryonal carcinoma (hEC) cells. Upon western blotting with R-10G, hiPS/ES cel...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cws159

    authors: Kawabe K,Tateyama D,Toyoda H,Kawasaki N,Hashii N,Nakao H,Matsumoto S,Nonaka M,Matsumura H,Hirose Y,Morita A,Katayama M,Sakuma M,Kawasaki N,Furue MK,Kawasaki T

    更新日期:2013-03-01 00:00:00

  • Impact of sialyltransferase ST6GAL1 overexpression on different colon cancer cell types.

    abstract::Cancer-associated glycan structures can be both tumor markers and engines of disease progression. The structure Siaα2,6Galβ1,4GlcNAc (Sia6LacNAc), synthesized by sialyltransferase ST6GAL1, is a cancer-associated glycan. Although ST6GAL1/Sia6LacNAc are often overexpressed in colorectal cancer (CRC), their biological an...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwz053

    authors: Venturi G,Gomes Ferreira I,Pucci M,Ferracin M,Malagolini N,Chiricolo M,Dall'Olio F

    更新日期:2019-09-20 00:00:00

  • Subunit-specific sulphation of oligosaccharides relating to charge-heterogeneity in porcine lutrophin isoforms.

    abstract::Lutrophin (LH) consists of an array of isoforms with different charges and bioactivities. This study was undertaken to clarify specifically how oligosaccharides of alpha and beta subunits contribute to LH isoform charges. Porcine LH (pLH) was separated into four isoforms by isoelectric focusing (IEF), followed by subu...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/2.3.225

    authors: Ujihara M,Yamamoto K,Nomura K,Toyoshima S,Demura H,Nakamura Y,Ohmura K,Osawa T

    更新日期:1992-06-01 00:00:00

  • A novel sialylated N-acetylgalactosamine-containing oligosaccharide is the major complex-type structure present in Bowes melanoma tissue plasminogen activator.

    abstract::We have employed fast atom bombardment mass spectrometry (FAB-MS) to screen the N-linked oligosaccharides of Bowes melanoma tissue plasminogen activator (mt-PA), and recombinant t-PAs produced by Chinese hamster ovary cells (rt-PA) and by a gene-enriched melanoma cell line (rmt-PA). These studies have confirmed the pu...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/1.2.173

    authors: Chan AL,Morris HR,Panico M,Etienne AT,Rogers ME,Gaffney P,Creighton-Kempsford L,Dell A

    更新日期:1991-03-01 00:00:00

  • The annotation of the asparagine N-linked glycosylation pathway in the Reactome database.

    abstract::Asparagine N-linked glycosylation is one of the most important forms of protein post-translational modification in eukaryotes and is one of the first metabolic pathways described at a biochemical level. Here, we report a new annotation of this pathway for the Human species, published after passing a peer-review proces...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwq215

    authors: Dall'Olio GM,Jassal B,Montanucci L,Gagneux P,Bertranpetit J,Laayouni H

    更新日期:2011-11-01 00:00:00

  • Carbohydrate recognition factors of a Talpha (Galbeta1-->3GalNAcalpha1-->Ser/Thr) and Tn (GalNAcalpha1-->Ser/Thr) specific lectin isolated from the seeds of Artocarpus lakoocha.

    abstract::Artocarpus lakoocha agglutinin (ALA), isolated from the seeds of A. lakoocha fruit, is a galactose-binding lectin and a potent mitogen of T and B cells. Knowledge obtained from previous studies on the affinity of ALA was limited to molecular and submolecular levels of Galbeta1-->3GalNAc (T) and its derivatives. In the...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh144

    authors: Singh T,Chatterjee U,Wu JH,Chatterjee BP,Wu AM

    更新日期:2005-01-01 00:00:00

  • Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation.

    abstract::Glycosyltransferases, usually residing within the intracellular secretory apparatus, also circulate in the blood. Many of these blood-borne glycosyltransferases are associated with pathological states, including malignancies and inflammatory conditions. Despite the potential for dynamic modifications of glycans on dis...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cww108

    authors: Lee-Sundlov MM,Ashline DJ,Hanneman AJ,Grozovsky R,Reinhold VN,Hoffmeister KM,Lau JT

    更新日期:2017-01-01 00:00:00

  • Digalactosylceramide is the receptor for staphylococcal enterotoxin-B in human kidney proximal tubular cells.

    abstract::We have characterized a glycosphingolipid (GSL) receptor for Staphylococcus enterotoxin-B (SEB) in cultured human kidney proximal tubular (PT) cells. Solid-phase binding of [125I]SEB to the GSL receptor was concentration dependent and was not displaceable by two structurally related toxins, such as staphylococcal ente...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/5.3.327

    authors: Chatterjee S,Khullar M,Shi WY

    更新日期:1995-05-01 00:00:00

  • Concanavalin A binding to HIV envelope protein is less sensitive to mutations in glycosylation sites than monoclonal antibody 2G12.

    abstract::Many mannose-binding proteins inhibit divergent strains of human immunodeficiency virus type 1 (HIV-1) in in vitro models of viral infectivity, suggesting that targeting mannose residues in vaccine applications might offset the strain restriction typically observed in antibody responses to HIV vaccine preparations. Co...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwi083

    authors: Pashov A,MacLeod S,Saha R,Perry M,VanCott TC,Kieber-Emmons T

    更新日期:2005-10-01 00:00:00

  • Addition of beta1-6 GlcNAc branching to the oligosaccharide attached to Asn 772 in the serine protease domain of matriptase plays a pivotal role in its stability and resistance against trypsin.

    abstract::beta1-6 GlcNAc branching, a product of N-acetylglucosaminyltransferase V (GnT-V), is a key structure that is associated with malignant transformations and cancer metastasis. Although a number of reports concerning tumor metastasis-related glycoproteins that contain beta1-6 GlcNAc branching have appeared, the precise f...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwh013

    authors: Ihara S,Miyoshi E,Nakahara S,Sakiyama H,Ihara H,Akinaga A,Honke K,Dickson RB,Lin CY,Taniguchi N

    更新日期:2004-02-01 00:00:00

  • Preparation and application of biologically active fluorescent hyaluronan oligosaccharides.

    abstract::We report the production of biologically active hyaluronan (HA) oligosaccharides labeled with the fluorophore 2-aminobenzoic acid (2AA). Oligosaccharides from 4 to 40 residues in length were purified to homogeneity by ion exchange chromatography using a logarithmic gradient. Molecular weight and purity characterizatio...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwi008

    authors: Seyfried NT,Blundell CD,Day AJ,Almond A

    更新日期:2005-03-01 00:00:00

  • Elimination of β-mannose glycan structures in Pichia pastoris.

    abstract::The methylotrophic yeast, Pichia pastoris, is an important organism used for the production of therapeutic proteins. However, the presence of fungal-like glycans, such as those containing β-mannose (Man) linkages, can elicit an immune response or bind to Man receptors, thus reducing their efficacy. Recent studies have...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwr108

    authors: Hopkins D,Gomathinayagam S,Rittenhour AM,Du M,Hoyt E,Karaveg K,Mitchell T,Nett JH,Sharkey NJ,Stadheim TA,Li H,Hamilton SR

    更新日期:2011-12-01 00:00:00

  • Structural characterization of the N-linked oligosaccharides in bile salt-stimulated lipase originated from human breast milk.

    abstract::The detailed structures of N- glycans derived from bile salt-stimulated lipase (BSSL) found in human milk were determined by combining exoglycosidase digestion with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The N- glycan structures were conclusively determined in terms of complexity...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/9.3.227

    authors: Mechref Y,Chen P,Novotny MV

    更新日期:1999-03-01 00:00:00

  • Identification of serum glycoprotein ligands for the immunomodulatory receptor blood dendritic cell antigen 2.

    abstract::Blood dendritic cell antigen 2 (BDCA-2) is a C-type lectin found on the surface of plasmacytoid dendritic cells. It functions as a glycan-binding receptor that downregulates the production of type I interferons and thus plays a role in oligosaccharide-mediated immunomodulation. The carbohydrate recognition domain in B...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwy050

    authors: Kim JW,Budzak J,Liu Y,Jégouzo SAF,Drickamer K,Taylor ME

    更新日期:2018-08-01 00:00:00

  • MUC2 mucin is a major carrier of the cancer-associated sialyl-Tn antigen in intestinal metaplasia and gastric carcinomas.

    abstract::Changes in mucin protein expression and in glycosylation are common features in pre-neoplastic lesions and cancer and are therefore used as cancer-associated markers. De novo expression of intestinal mucin MUC2 and cancer-associated sialyl-Tn antigen are frequently observed in intestinal metaplasia (IM) and gastric ca...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwp161

    authors: Conze T,Carvalho AS,Landegren U,Almeida R,Reis CA,David L,Söderberg O

    更新日期:2010-02-01 00:00:00

  • Temperature-dependent cooperativity in donor-acceptor substrate binding to the human blood group glycosyltransferases.

    abstract::Affinities of the human blood group glycosyltransferases, alpha-(1-->3)-N-acetylgalactosaminyltransferase (GTA) and alpha-(1-->3)-galactosyltransferase (GTB) for their common acceptor substrate alpha-l-Fucp-(1-->2)-beta-d-Galp-O(CH2)(7)CH3 (1), in the absence and presence of bound uridine 5'-diphosphate (UDP) and Mn2+...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwn043

    authors: Shoemaker GK,Soya N,Palcic MM,Klassen JS

    更新日期:2008-08-01 00:00:00

  • N-linked oligosaccharide processing enzyme glucosidase II produces 1,5-anhydrofructose as a side product.

    abstract::alpha-1,4-Glucan lyase cleaves alpha-1,4-linkages of nonreducing termini of alpha-1,4-glucans to produce 1,5-anhydrofructose (1,5-AnFru). The enzymes isolated from fungi and algae show high homology with glycoside hydrolase family 31. Purification of alpha-1,4-glucan lyase from rat liver using DEAE Cellulose chromatog...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/10.12.1283

    authors: Hirano K,Ziak M,Kamoshita K,Sukenaga Y,Kametani S,Shiga Y,Roth J,Akanuma H

    更新日期:2000-12-01 00:00:00

  • Processing inhibition of N-linked sugar chains associated with induction of Fc receptor-mediated phagocytosis in the mouse monocytoid cells.

    abstract::The concanavalin A staining of cellular glycoproteins and the direct analysis of their sugar chains released by hydrazinolysis revealed that the processing of N-linked sugar chains of some glycoproteins is suppressed by exposure of mouse monocytoid cells P388D1 to dimethyl sulphoxide, which can induce Fc receptor-medi...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/3.1.15

    authors: Fukushima K,Takasaki S

    更新日期:1993-02-01 00:00:00

  • Cell cycle-specific growth inhibition of human breast cancer cells induced by metabolic inhibitors.

    abstract::Proliferation of exponentially growing breast cancer cells (line Hs578T) was blocked specifically in G1 by 3-hydroxy-3-methylglutaryl Coenzyme A (HMG CoA) reductase inhibition, as well as by inhibition of N-linked glycosylation. As a consequence of these inhibitory conditions, the cells were synchronized in the G1 sta...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/3.5.475

    authors: Larsson O

    更新日期:1993-10-01 00:00:00

  • Glycosylation and biological activity of CAMPATH-1H expressed in different cell lines and grown under different culture conditions.

    abstract::CAMPATH-1H (where CAMPATH is a trade mark of Wellcome group companies), a humanized IgG antibody used in the therapy of lymphoma, leukaemia and rheumatoid arthritis, has been expressed in Chinese hamster ovary, Y0 myeloma and NS0 myeloma cell lines. These engineered cell lines were grown under different culture condit...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/5.8.813

    authors: Lifely MR,Hale C,Boyce S,Keen MJ,Phillips J

    更新日期:1995-12-01 00:00:00

  • Expression regulation and function of heparan sulfate 6-O-endosulfatases in the spermatogonial stem cell niche.

    abstract::Glial cell line-derived neurotrophic factor (GDNF) is a heparan sulfate (HS)-binding factor. GDNF is produced by somatic Sertoli cells, where it signals to maintain spermatogonial stem cells (SSCs) and reproduction. Here, we investigate the roles of extracellular HS 6-O-endosulfatases (Sulfs), Sulf1 and Sulf2, in the ...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/cwq133

    authors: Langsdorf A,Schumacher V,Shi X,Tran T,Zaia J,Jain S,Taglienti M,Kreidberg JA,Fine A,Ai X

    更新日期:2011-02-01 00:00:00

  • Characterization of the oligosaccharides assembled on the Pichia pastoris-expressed recombinant aspartic protease.

    abstract::Aspartic protease, widely used as a milk-coagulating agent in industrial cheese production, contains three potential N-glycosylation sites. In this study, we report the characterization of N-linked oligosaccharides on recombinant aspartic protease secreted from the methylotrophic yeast Pichia pastoris using a combinat...

    journal_title:Glycobiology

    pub_type: 杂志文章

    doi:10.1093/glycob/9.10.1037

    authors: Montesino R,Nimtz M,Quintero O,García R,Falcón V,Cremata JA

    更新日期:1999-10-01 00:00:00