Probing multivalent carbohydrate–lectin interactions by an enzyme-linked lectin assay employing covalently immobilized carbohydrates

Maierhofer, Caroline; Rohmer, Katja; Wittmann, Valentin

doi:10.1016/j.bmc.2007.08.063

Cited by 49 publications

(41 citation statements)

References 62 publications

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“…To this end, inhibition of horseradish-peroxidase (HRP)-labeled WGA binding to an immobilized GlcNAc ligand was determined as previously described. [19] The results are shown in Table 1. The free sugar GlcNAc inhibited the WGA binding with an IC 50 of 30 mm.…”

Section: Binding Experiments With Wgamentioning

confidence: 96%

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Rapid Screening of Lectins for Multivalency Effects with a Glycodendrimer Microarray

et al. 2010

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Multivalency is an important phenomenon in protein-carbohydrate interactions. In order to evaluate glycodendrimers as multivalent inhibitors of carbohydrate binding proteins, we displayed them on a microarray surface. Valencies were varied from 1 to 8, and corrections were made for the valencies so that all surfaces contained the same amount of the sugar ligand. Five different carbohydrates were attached to the dendrimers. A series of fluorescent lectins was evaluated, and for each of them a binding profile was obtained from a single experiment showing both the specificity of the lectin for a certain sugar and whether it prefers multivalent ligands or not. Very distinct binding patterns were seen for the various lectins. The results were rationalized with respect to the interbinding distances of the lectins.

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Section: Binding Experiments With Wgamentioning

confidence: 96%

“…Very strong multivalency effects of up to factors of 10 6 have been measured with bacterial toxins and other multisite lectins. [10,18,19] Recently we took a step to study multivalency effects more efficiently by using a microarray. Microarrays are increasingly being explored in the study of protein-carbohydrate interactions.…”

Section: Introductionmentioning

confidence: 99%

Rapid Screening of Lectins for Multivalency Effects with a Glycodendrimer Microarray

et al. 2010

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“…WGA-sugar interactions have been revealed by means of NMR 21 and mass spectrometry. 22 Because the WGA-sugar binding in living bodies is interesting, monitoring of the binding has been carried out using enzyme-linked immunosorbent assay 23 and electrophoresis. 24 Our voltammetric procedure to evaluate the binding does not require separation of free WGA from the bound WGA before measurement.…”

Section: Effect Of Wga Incubation Time On Thionine/glucose-modified Mmentioning

confidence: 99%

Voltammetric Evaluation of the Binding between Wheat Germ Agglutinin and Thionine/Glucose-modified Magnetic Microbeads

et al. 2008

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Cells are the fundamental structural and functional units of living organisms. The cell membrane is comprised of phospholipid and gycolipids; glycoproteins are located on the cell membrane surface. The sugar residues of glycoproteins combine with biological molecules such as antibodies, hormones and enzymes. [1][2][3] Bacteria and viruses also recognize the structure and sequence of sugar residues and invade the host by binding to them. 4 Lectins are proteins that interact with specific sugar residues. Binding between lectin and sugar residues causes cell agglutination and precipitation of polysaccharides. 5Because many lectins are commercially available and are easy to handle, they are well-suited for the study of protein-sugar residue binding. 6 For example, one type of lectin wheat germ agglutinin (WGA) is a 36000 molecular weight protein consisting of two identical sub-units.WGA specifically binds the sugar, N-acetylglucosamine, which is comprised of glucosamine and glucose. Generally, the terminal sugar residue on the cell surface is either glucose, N-acetylglucosamine, mannose or xylose. Therefore, the binding between WGA and glucose plays an important role in various physiological processes. 7 For example, the time dependence of agglutination and the spreading of human erythrocytes induced by cell-cell contact involve WGA-glucose binding. 8 The importance of WGA-glucose binding for the sexual development of dictyostelium discoideum also has been reported. 9WGA-glucose has also been applied to affinity chromatography, using glycoconjugates and WGA as the ligands. 10Various models of biological cells have been used to study living cells. Bilayer lipid membranes are widely used because they replicate the electric and hydrophobic properties of the cell membrane.11 Liposomes also have been used as a model of the cell.12 Magnetic microbeads are the same size as living cells; thus, magnetic microbeads are used as cell models in medical, biological and environmental research. Furthermore, functional molecules can be immobilized onto the surface of magnetic beads via cross-linking. The beads can be easily isolated from solutions containing protein or from bacterial cultures by application of a magnetic field.Magnetic beads have been used in a variety of immunologic and electrochemical assays. Kim et al. detected abnormal prion proteins using a magnetic bead-based immunoassay coupled with laser-induced fluorescence spectrofluorometry. 13 In addition, immunomagnetic beads have been used in the detection of bacillus stearothermophilus spores in food and environmental samples.14 Microbeads with fluorogenic 15 and chemiluminescence reagents 16 have been developed to evaluate protein-sugar residue binding. Thomas et al. established an electrochemical procedure for the evaluation of the antigen-antibody reaction on the magnetic bead surface using an enzyme reaction. 17 Our group previously reported a method for the evaluation of lectin-sugar binding on sugar-modified beads. 18 In this method, the sugar was labeled with a...

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“…Há diversos relatos em que dímeros, trímeros e tetrâmeros apresentam afinidade superior, quando comparados a análogos de maior valência. [15][16][17][18] Esse fato, aliado à simplicidade estrutural e baixo peso molecular dos dímeros, tem despertado o interesse em sua síntese. [18][19][20][21][22] .Recentemente relatamos a síntese de uma série de ligantes monoméricos derivados de D-galactose e lactose (4-O-(b-Dgalactopiranosil)-D-glicose) e a interação desses com a lectina de Erythrina cristagalli (LEC).…”

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Síntese de β-D-galactopiranosídeos de arila diméricos para avaliação de sua interação com a lectina de Erythrina cristagalli

Figueiredo¹,

Meyer²,

Prado³

et al. 2009

Quím. Nova

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Recebido em 31/12/08; aceito em 13/4/09; publicado na web em 22/9/09 SYNTHESIS OF DIMERIC ARYL b-D-GALACTOPYRANOSIDES FOR THE EVALUATION OF THEIR INTERACTION WITH THE Erythrina cristagalli LECTIN.The synthesis of two new D-galactose-based dimers having a 1,4-butanediamine spacer is reported aiming at the evaluation of their interaction with the Erythrina cristagalli lectin. The title compounds were prepared in four and five steps from 2,3,4,6-tetra-O-acetyl-b-D-galactopyranoside bromide, in 20 % and 15 % overall yield, respectively, using the Doebner modification of the Koenavenagel reaction as the key sep. The lectin-carbohydrate interaction could be evaluated for only one dimer, due to solubility problems. A twofold enhancement of affinity was observed, compared to the corresponding monovalent ligand.Keywords: D-galactopyranose; glycodendrimers; Erythrina cristagalli. INTRODUÇÃOOs glicoconjugados como as glicoproteínas e os glicolipídios possuem um papel importante em vários processos biológicos relacionados com reconhecimento e adesão celular, como inflamação, infecção e mestástase.1,2 As forças que regem esses processos de reconhecimento e adesão são interações complexas entre as porções carboidrato de um glicoconjugado e as lectinas, proteínas que reconhecem especificamente carboidratos.Usualmente, a afinidade das lectinas por monossacarídeos é fraca. Entretanto, na natureza, essas interações são multivalentes. A multivalência está associada ao reconhecimento de múltiplas cópias de carboidrato apresentadas a múltiplas cópias de lectinas presentes na superfície celular.3 A multivalência aumenta a força e a seletividade das interações carboidrato-lectinas. 4 Substâncias multivalentes sintéticas derivadas de carboidratos, denominadas genericamente glicodendrímeros, 4 mimetizam a interação dos glicoconjugados com lectinas no sistema biológico. Há, portanto, grande interesse na obtenção dessas substâncias, para avaliação de suas interações com lectinas.1,4-8 Tem-se demonstrado que a eficiência das interações entre glicodendrímeros e lectinas depende de vários fatores, como o número de unidades de carboidratos presentes na molécula (multivalência), a distância entre essas unidades e a flexibilidade do dendrímero. [9][10][11] De modo geral, aumentando-se a multivalência aumenta-se a eficiência da interação carboidratolectina.12-14 Entretanto, isso nem sempre é o caso. Há diversos relatos em que dímeros, trímeros e tetrâmeros apresentam afinidade superior, quando comparados a análogos de maior valência. [15][16][17][18] Esse fato, aliado à simplicidade estrutural e baixo peso molecular dos dímeros, tem despertado o interesse em sua síntese. [18][19][20][21][22] .Recentemente relatamos a síntese de uma série de ligantes monoméricos derivados de D-galactose e lactose (4-O-(b-Dgalactopiranosil)-D-glicose) e a interação desses com a lectina de Erythrina cristagalli (LEC). 23 No presente trabalho é descrita a síntese de dois dímeros derivados de D-galactose, com o mesmo objetivo. Esses dímeros (estruturas 6 e 7, Figura ...

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Probing multivalent carbohydrate–lectin interactions by an enzyme-linked lectin assay employing covalently immobilized carbohydrates

Cited by 49 publications

References 62 publications

Rapid Screening of Lectins for Multivalency Effects with a Glycodendrimer Microarray

Rapid Screening of Lectins for Multivalency Effects with a Glycodendrimer Microarray

Voltammetric Evaluation of the Binding between Wheat Germ Agglutinin and Thionine/Glucose-modified Magnetic Microbeads

Síntese de β-D-galactopiranosídeos de arila diméricos para avaliação de sua interação com a lectina de Erythrina cristagalli

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