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 characterization of HA oligosaccharides is facilitated by 2AA derivatization because it enhances signals in MALDI-TOF MS and improves FACE (fluorophore-assisted carbohydrate electrophoresis) analysis by avoiding the inverted parabolic migration characteristic of 2-aminoacridone (AMAC)-labeled sugars. The small size and shape of the fluorophore maintains the biological activity of the derivatized oligosaccharides, as demonstrated by their ability to compete for polymeric HA binding to the G1-domain of human recombinant versican (VG1). An electrophoretic mobility shift assay was used to study VG1 binding to labeled HA 8-, 10-, 20-, 30-, and 40-mers, and although no stable VG1 binding was observed to labeled 8-mers, the equilibrium dissociation constant (100 nM) for VG1 with HA(10) was estimated from densitometry analysis of the free oligosaccharide. Interactions involving HA 20-, 30-, and 40-mers (proposed to be multivalent) could also be studied using this protocol. Oligosaccharides labeled with 2AA therefore show excellent potential as probes in fluorescence-based assays that investigate protein-carbohydrate interactions.
journal_name
Glycobiologyjournal_title
Glycobiologyauthors
Seyfried NT,Blundell CD,Day AJ,Almond Adoi
10.1093/glycob/cwi008subject
Has Abstractpub_date
2005-03-01 00:00:00pages
303-12issue
3eissn
0959-6658issn
1460-2423pii
cwi008journal_volume
15pub_type
杂志文章相关文献
GLYCOBIOLOGY文献大全abstract::The transcriptional regulation mechanisms involved in the up-regulation of Fas-induced GD3 synthase gene have not yet been elucidated. 5'-Rapid amplification of cDNA end (5'-RACE) using mRNA prepared from Fas-induced Jurkat T cells revealed the presence of multiple transcription start sites of human GD3 synthase gene,...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwj087
更新日期:2006-05-01 00:00:00
abstract::Sialidases are widely distributed glycohydrolytic enzymes removing sialic acid residues from glycoconjugates. In mammals, several sialidases with different subcellular localizations and biochemical features have been described. NEU4, the most recently identified member of the human sialidase family, is found in two fo...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwp156
更新日期:2010-02-01 00:00:00
abstract::Mycobacterium tuberculosis lipoarabinomannan (LAM) and biosynthetically related lipoglycans and glycans play an important role in host-pathogen interactions. Therefore, the elucidation of the complete biosynthetic pathways of these important molecules is expected to afford novel therapeutic targets. The characterizati...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwv061
更新日期:2015-11-01 00:00:00
abstract::Lepidopteran insect cells are used routinely as hosts for foreign glycoprotein expression by recombinant baculoviruses, but the precise nature of their N-glycosylation pathway remains poorly defined. These cells clearly have processing glucosidases and mannosidases that can convert precursors to Man3GlcNAc2 structures...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/7.1.113
更新日期:1997-02-01 00:00:00
abstract::The enzyme UDP-N-acetylglucosamine: α-d-mannoside β-1-6 N-acetylglucosaminyltransferase V (GnT-V) catalyzes the transfer of GlcNAc from the UDP-GlcNAc donor to the α-1-6-linked mannose of the trimannosyl core structure of glycoproteins to produce the β-1-6-linked branching of N-linked oligosaccharides. β-1-6-GlcNAc-br...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cww010
更新日期:2016-07-01 00:00:00
abstract::Human milk oligosaccharides (HMOS) are not digested in the proximal intestine. In distal intestine, HMOS collectively modify the microbiota, but the response of individual bacteria to individual components of the HMOS is not well defined. Here, each of 25 major isolates of the human intestinal microbiota was fed indiv...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwt065
更新日期:2013-11-01 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::In cancer cells, the glycoprotein Mucin 1 (MUC1) undergoes abnormal, truncated glycosylation. The truncated glycosylation exposes cryptic peptide epitopes that can be recognized by antibodies. Since these immunogenic regions are cancer specific, they represent ideal targets for therapeutic antibodies. We investigated ...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cww131
更新日期:2017-07-01 00:00:00
abstract::The transplantation of organs from lower animals such as pigs into humans is prevented by a severe rejection reaction initiated by complement fixing xenoreactive natural antibodies. Most anti-pig xenoreactive natural antibodies in humans are thought to recognize Gal alpha 1-3Gal beta 1-4GlcNAc and are also thought to ...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/6.5.499
更新日期:1996-07-01 00:00:00
abstract::Galectin 1 (GAL1) is a beta-galactoside-binding lectin involved in cell cycle progression. GAL1 overexpression is associated with neoplastic transformation and loss of differentiation. The gene encoding for human GAL1 resides on chromosome 22(q12; q13), and its expression is developmentally regulated. Although devoid ...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/7.8.1193
更新日期:1997-12-01 00:00:00
abstract::We report here a method of simultaneously quantifying glucosylceramide (GlcCer) and galactosylceramide (GalCer) by normal-phase HPLC using O-phtalaldehyde derivatives. Treatment with sphingolipid ceramide N-deacylase which converts the cerebrosides in the sample to their lyso-forms was followed by the quantitative lab...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwp047
更新日期:2009-07-01 00:00:00
abstract::The catabolism of dermatan sulfate (DS) commences with endohydrolysis of the polysaccharide to oligosaccharides by proposed endo-beta-N-acetylhexosaminidase and endohexuronidase activities. To investigate the substrate specificities of these activities, we developed an assay to measure specific products of their actio...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwn097
更新日期:2008-12-01 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::Gangliosides (sialic acid-containing glycosphingolipids) play important roles in many physiological functions, including synaptic plasticity in the hippocampus, which is considered as a cellular mechanism of learning and memory. In the present study, three types of synaptic plasticity, long-term potentiation (LTP), lo...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwr090
更新日期:2011-10-01 00:00:00
abstract::Helicobacter pylori infects over half of the world's population and is thought to be a leading cause of gastric ulcer, gastric carcinoma, and gastric malignant lymphoma of mucosa-associated lymphoid tissue type. Previously, we reported that a gland mucin (MUC6) present in the lower portion of the gastric mucosa contai...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwn037
更新日期:2008-07-01 00:00:00
abstract::Heparin/heparan sulfates (H/HS) are ubiquitous biopolymers that interact with many proteins to induce a range of biological functions. Unfortunately, how these biopolymers recognize their preferred protein targets remain poorly understood. It is suggested that computational simulations offer attractive avenues but a n...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwaa015
更新日期:2020-07-16 00:00:00
abstract::Rod and cone photoreceptors project from the outer retinal surface into a carbohydrate-rich interphotoreceptor matrix (IPM). Unique IPM glycoconjugates are distributed around rods and cones. Wheat germ agglutinin (WGA) strongly decorates the rod matrix domains and weakly decorates the cone matrix domains. This study c...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/8.10.997
更新日期:1998-10-01 00:00:00
abstract::Human alpha-galactosidase A (alpha-Gal A) is the lysosomal glycohydrolase that cleaves the terminal alpha-galactosyl moieties of various glycoconjugates. Overexpression of the enzyme in Chinese hamster ovary (CHO) cells results in high intracellular enzyme accumulation and the selective secretion of active enzyme. Str...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/8.4.329
更新日期:1998-04-01 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::Intelectin is an extracellular animal lectin found in chordata. Although human and mouse intelectin-1 recognize galactofuranosyl residues included in cell walls of various microorganisms, the physiological function of mammalian intelectin had been unclear. In this study, we found that human intelectin-1 was a serum pr...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwp013
更新日期:2009-05-01 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::Recombinant human erythropoietin (rHuEPO) is used abundantly in the clinic to stimulate red blood cell growth in anaemic patients. The efficacy of the drug depends strongly on the extent of sialylation of its carbohydrate moiety. Prompted by conflicting literature reports on the issue, we reinvestigated the structures...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/4.2.227
更新日期:1994-04-01 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::Lectins are used as defense effector proteins against predators, parasites and pathogens by animal, plant and fungal innate defense systems. These proteins bind to specific glycoepitopes on the cell surfaces and thereby interfere with the proper cellular functions of the various antagonists. The exact cellular toxicit...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cww113
更新日期:2017-05-01 00:00:00
abstract::Schistosomiasis caused by infection with parasitic helminths of Schistosoma spp. is a major global health problem due to inadequate treatment and lack of a vaccine. The immune response to schistosomes includes glycan antigens, which could be valuable diagnostic markers and vaccine targets. However, no precedent exists...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwu027
更新日期:2014-07-01 00:00:00
abstract::Since the turn of the century, cancer therapy has undergone a transformation in terms of new treatment modalities and renewed optimism in achieving long-lived tumor control and even cure. This is, in large part, thanks to the widespread incorporation of monoclonal antibodies (mAbs) into standard treatment regimens. Th...
journal_title:Glycobiology
pub_type: 杂志文章,评审
doi:10.1093/glycob/cwy043
更新日期:2018-09-01 00:00:00
abstract::Galectin-1, the prototype of a family of beta-galactoside-binding proteins, has been implicated in a wide variety of biological processes. Data presented herein show that galectin-1 stimulates monocyte migration in a dose-dependent manner but is not chemotactic for macrophages. Galectin-1-induced monocyte chemotaxis i...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwp077
更新日期:2009-12-01 00:00:00
abstract::The glycobiology of Schistosoma mansoni is dominated by developmentally regulated expression of various fucosylated structures, most notably the Lewis X epitope and a multifucosylated sequence, Fuc alpha1-->2Fuc alpha1-->, in its various forms. For the infective cercarial stage, Lewis X has been structurally identifie...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/11.5.395
更新日期:2001-05-01 00:00:00
abstract::Schistosoma mansoni soluble egg antigens (SEAs) are crucially involved in modulating the host immune response to infection by S. mansoni. We report that human dendritic cells bind SEAs through the C-type lectin dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN). Monoclonal antibodies against the carbohydrat...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwg052
更新日期:2003-06-01 00:00:00
abstract::We have recently demonstrated that the mannose-binding lectins, namely banana lectin (BL) and garlic lectin (GL), interacted with the insulin receptors on M210B4 cells--an established mesenchymal cell line of murine marrow origin--and initiate mitogen-activated protein kinase kinase (MEK)-dependent extracellular signa...
journal_title:Glycobiology
pub_type: 杂志文章
doi:10.1093/glycob/cwq194
更新日期:2011-04-01 00:00:00