Abstract:
:We recently described a novel fluorescent compound, 2-amino,6-amidobiotinyl-pyridine (BAP), that allows the tagging of oligosaccharides, their fractionation by reversed-phase HPLC with picomole scale detection, and the formation of functional neoglycoprotein equivalents with (strept) avidin for the detection of receptors and the generation of monospecific antibodies (Rothenberg et al., Proc. Natl Acad. Sci. USA, 90, 11939-11943, 1993). Here, we describe the enhancement of this approach by the following. (i) A simple one-step purification of BAP from its synthetic precursors and other reactants. (ii) Development of HPLC sizing column methods to quickly purify BAP-coupled oligosaccharides away from free BAP and other reactants. (iii) Development of anion-exchange and amine-adsorption HPLC procedures for the fractionation of BAP-oligosaccharide adducts by charge and size, respectively. (iv) Investigation of the affinity of BAP-oligosaccharides for (strept)avidin, confirming the formation of stable complexes. (v) The use of BAP for sensitive monosaccharide compositional analysis of glycoproteins. (vi) Formation of stable BAP adducts without reduction and its implications for the mechanism of adduct formation. These advances make available a multitude of techniques for the fractionation of BAP-coupled oligosaccharides based on several different physical parameters. Distinct species of BAP-coupled oligosaccharides can be isolated and subjected to detailed structural analysis. Such defined molecules form stable complexes with streptavidin that are effectively neoglycoproteins, which can be used in a variety of biological applications. Notably, all of these approaches require relatively inexpensive materials and conventional equipment available in most laboratories.
journal_name
Glycobiologyjournal_title
Glycobiologyauthors
Toomre DK,Varki Adoi
10.1093/glycob/4.5.653subject
Has Abstractpub_date
1994-10-01 00:00:00pages
653-63issue
5eissn
0959-6658issn
1460-2423journal_volume
4pub_type
杂志文章相关文献
GLYCOBIOLOGY文献大全abstract::The human anticoagulant factor, Protein C, is a plasma glycoprotein that has reported anti-ischaemic and anti-inflammatory properties. To explore potential mechanisms for these reported activities, we examined the effect of Protein C on the process of cell adhesion to vascular endothelial cells, which plays a critical...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/4.2.221
更新日期:1994-04-01 00:00:00
abstract::Core alpha1,6-fucosylation is a conserved feature of animal N-linked oligosaccharides being present in both invertebrates and vertebrates. To prove that the enzymatic basis for this modification is also evolutionarily conserved, cDNAs encoding the catalytic regions of the predicted Caenorhabditis elegans and Drosophil...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwi028
更新日期:2005-05-01 00:00:00
abstract::Since plants are emerging as an important system for the expression of recombinant glycoproteins, especially those intended for therapeutic purposes, it is important to scrutinize to what extent glycans harbored by mammalian glycoproteins produced in transgenic plants differ from their natural counterpart. We report h...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/9.4.365
更新日期:1999-04-01 00:00:00
abstract::Heparan sulfate is a polysaccharide belonging to the glycaminoglycan family. It interacts with numerous proteins of the extracellular matrix, in particular cellular growth factors. The number of experimental protein-heparin sulfate complexes obtained by crystallography or nuclear magnetic resonance is limited. Alterna...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwr052
更新日期:2011-09-01 00:00:00
abstract::The structure of the O-antigen polysaccharide (PS) of Escherichia coli O115 has been investigated using a combination of component analysis and 1D and 2D nuclear magnetic resonance (NMR) spectroscopy experiments. The repeating unit of the O-antigen was elucidated using the O-deacetylated PS and has the following branc...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cws161
更新日期:2013-03-01 00:00:00
abstract::A glycosyltransferase was fused to the yeast cell wall protein Pir, which forms the Pir1-4 protein family and is incorporated into the cell wall by an unknown linkage to be displayed at the yeast cell surface. We first expressed the PIR1-HA-gma12+ fusion, in which gma12+ encodes alpha-1,2-galactosyltransferase from th...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwg014
更新日期:2003-02-01 00:00:00
abstract::Viscumin of mistletoe (Viscum album L.) has a concentration-dependent activity profile unique to plant AB-toxins. It starts with lectin-dependent mitogenicity and then covers toxicity and cell agglutination, associated with shifts in the monomer/dimer equilibrium. Each lectin subunit harbors two sections for ligand co...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwl017
更新日期:2006-10-01 00:00:00
abstract::A common argument against using plants as a production system for therapeutic proteins is their inability to perform authentic N-glycosylation. A major concern is the presence of beta 1,2-xylose and core alpha 1,3-fucose residues on complex N-glycans as these nonmammalian N-glycan residues may provoke unwanted side ef...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwm137
更新日期:2008-03-01 00:00:00
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
更新日期:2013-05-01 00:00:00
abstract::Galectins are a sub-family of lectins, defined by their highly conserved beta-sandwich structures and ability to bind to beta-galactosides, like Gal beta1-4 Glc (lactose). Here, we used (15)N-(1)H HSQC and pulse field gradient (PFG) NMR spectroscopy to demonstrate that galectin-1 (gal-1) binds to the relatively large ...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwp084
更新日期:2009-09-01 00:00:00
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
更新日期:1992-06-01 00:00:00
abstract::To facilitate deciphering the information content in the glycome, thin film-coated photoactivatable surfaces were applied for covalent immobilization of glycans, glycoconjugates, or lectins in microarray formats. Light-induced immobilization of a series of bacterial exopolysaccharides on photoactivatable dextran-coate...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwh143
更新日期:2005-01-01 00:00:00
abstract::Giardia lamblia, the protist that causes diarrhea, makes an Asn-linked-glycan (N-glycan) precursor that contains just two sugars (GlcNAc(2)) attached by a pyrophosphate linkage to a polyprenol lipid. Because the candidate cis-prenyltransferase of Giardia appears to be more similar to bacterial enzymes than to those of...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwq036
更新日期:2010-07-01 00:00:00
abstract::Isolation of glycopeptides utilizing hydrogen bonding between glycopeptide glycans and a carbohydrate-gel matrix in the organic phase is useful for site-specific characterization of oligosaccharides of glycoproteins, when combined with mass spectrometry. In this study, recovery of glycopeptides was improved by includi...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwj019
更新日期:2005-12-01 00:00:00
abstract::Human colon cancer is associated with antigenic and structural changes in mucin-type carbohydrate chains (O-glycans). To elucidate the control of the biosynthesis of these O-glycans is colon cancer, we have studied glycosyltransferase and sulphotransferase activities involved in the assembly of elongated O-glycan stru...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/4.6.873
更新日期:1994-12-01 00:00:00
abstract::Phytohaemagglutinin (PHA) is the major lectin present in the seeds of the common bean, Phaseolus vulgaris, and PHA-L is the leucocyte-agglutinating form of this lectin. This tetrameric glycoprotein accumulates in the vacuoles of storage parenchyma cells. Based on amino acid sequence comparisons of legume lectins and t...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/3.6.581
更新日期:1993-12-01 00:00:00
abstract::The structure of the complex mixture of the core oligosaccharide components of the lipooligosaccharide (LOS) fraction of Agrobacterium tumefaciens strain TT111 was determined directly on the deacetylated products by means of spectroscopical methods. The rhamnan oligosaccharide elongating the inner Kdo residue shares s...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwl033
更新日期:2006-12-01 00:00:00
abstract::Glycosylation is a common modification found on numerous proteins and lipids. However, direct detection of glycans on these intact biomolecules has been challenge. Here, utilizing enzymatic incorporation of fluorophore-conjugated sialic acids, dubbed as direct fluorescent glycan labeling, we report the labeling and de...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwz058
更新日期:2019-10-21 00:00:00
abstract::A protocol is described for uniform 13C labelling of terminal galactose residues of the glycan chains of glycoproteins, using an enzymatic method which does not perturb the protein. The technique is illustrated by application to the biantennary N-linked glycan chains attached at Asn 297 of immunoglobulin G (IgG). Isot...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/4.4.485
更新日期:1994-08-01 00:00:00
abstract::Lymphocytic beta 1,4-galactosyltransferase (beta 1,4-GalTase, EC 2.4.1.38) activity was measured in B cells using a neoglyco-protein, N-acetylglucosamine-phenylisothiocyanate-bovine serum albumin (GlcNAc-pITC-BSA), as an acceptor substrate in a novel enzyme-linked immunosorbent assay (ELISA)-based method. This assay p...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/5.4.365
更新日期:1995-06-01 00:00:00
abstract::N-acetylneuraminic acid (5-acetamido-3,5-dideoxy-d-glycero-d-galacto-non-2-ulosonic acid), which is the principal sialic acid family member of the non-2-ulosonic acids and their various derivatives, is often found at the terminal position on the glycan chains that adorn all vertebrate cells. This terminal position com...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwz099
更新日期:2020-04-20 00:00:00
abstract::Mouse macrophage galactose/N-acetylgalactosamine-specific C-type lectin (MMGL) is a type II transmembrane glycoprotein belonging to the C-type lectin family. Our development of monoclonal antibodies led us to discover that a calcium-dependent conformational change is detected by an antibody (termed mAb LOM-11) and tha...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/8.8.791
更新日期:1998-08-01 00:00:00
abstract::Heparan sulfates (HSs) exert critical regulatory actions on many proteins, including growth factors, and are essential for normal development. Variations in their specific sulfation patterns are known to regulate binding and signaling of fibroblast growth factors (FGFs) via tyrosine kinase receptors (FGFRs). We previo...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwf072
更新日期:2002-11-01 00:00:00
abstract::There has been a recent explosion in the identification of neuromuscular diseases caused by mutations in genes that affect carbohydrate metabolism or protein glycosylation. A number of these findings relate to defects in the glycosylation of alpha dystroglycan. Alpha dystroglycan is an essential component of the dystr...
journal_title:Glycobiology
pub_type: 杂志文章,评审
doi:10.1093/glycob/cwg077
更新日期:2003-08-01 00:00:00
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
更新日期:2005-10-01 00:00:00
abstract::Bacterial protein glycosylation systems from varying species have been functionally reconstituted in Escherichia coli. Both N- and O-linked glycosylation pathways, in which the glycans are first assembled onto lipid carriers and subsequently transferred to acceptor proteins by an oligosaccharyltransferase (OTase), hav...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cws059
更新日期:2012-07-01 00:00:00
abstract::Erythropoietin (Epo) is a 166 amino acids protein containing three N-glycosylation sites (Asn-24, Asn-38, and Asn-83) and 1 O- glycosylation site (Ser-126) and involved in the regulation of the level of red blood cells. Today, only one recombinant human Epo (rHuEpo), produced in CHO cell line, is extensively used in t...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/10.5.511
更新日期:2000-05-01 00:00:00
abstract::Poly-N-acetyllactosamine extensions on N- and O-linked glycans are increasingly recognized as biologically important structural features, but access to these structures has not been widely available. Here, we report a detailed substrate specificity and catalytic efficiency of the bacterial β3-N-acetylglucosaminyltrans...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cws101
更新日期:2012-11-01 00:00:00
abstract::In this study, we report the first cloning and characterization of a N-acetylneuraminic acid phosphate synthase gene from Drosophila melanogaster, an insect in the protostome lineage. The gene is ubiquitously expressed at all stages of Drosophila development and in Schneider cells. Similar to the human homologue, the ...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/12.2.73
更新日期:2002-02-01 00:00:00
abstract::UDP-N-acetylglucosamine (UDP-GlcNAc) is a glucose metabolite with pivotal functions as a key substrate for the synthesis of glycoconjugates like hyaluronan, and as a metabolic sensor that controls cell functions through O-GlcNAc modification of intracellular proteins. However, little is known about the regulation of h...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cww019
更新日期:2016-07-01 00:00:00