Low Affinity Carbohydrate Lectin Interactions Examined with Surface Plasmon Resonance

Weimar, Thomas; Haase, Bernd; Köhli, Thies

doi:10.1080/07328300008544136

Cited by 8 publications

(4 citation statements)

References 24 publications

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“…The lectins AAA and LTL-A were immobilized on a dextran chip, and the results are compiled in Table . The data were similar to K D values determined previously for l - fucose and for the disaccharide α - l - Fuc-(1→6)-β- d -GlcNAc-OMe ( 8 ) . These data are also included in Table .…”

Section: Resultssupporting

confidence: 84%

Molecular Recognition of Sialyl Lewis^xand Related Saccharides by Two Lectins

2001

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The interaction of sialyl Lewis(x), Lewis(x), and alpha-L-Fuc-(1-->3)-beta-D-GlcNAc with isolectin A from Lotus tetragonolobus (LTL-A), and with Aleuria aurantia agglutinin (AAA) was studied using NMR experiments and surface plasmon resonance. Both lectins are specific for fucose residues. From NMR experiments it was concluded that alpha-L-Fuc-(1-->3)-beta-D-GlcNAc and Lewis(x) bound to both lectins, whereas sialyl Lewis(x) only bound to AAA. Increased line broadening of 1H NMR signals of the carbohydrate ligands upon binding to AAA and LTL-A suggested that AAA bound to the ligands more tightly. Further comparison of line widths showed that for both lectins binding strengths decreased from alpha-L-Fuc-(1-->3)-beta-D-GlcNAc to Lewis(x) and were lowest for sialyl Lewis(x). Surface plasmon resonance measurements were then employed to yield accurate dissociation constants. TrNOESY, QUIET-trNOESY, and trROESY experiments delivered bioactive conformations of the carbohydrate ligands, and STD NMR experiments allowed a precise epitope mapping of the carbohydrates bound to the lectins. The bioactive conformation of Lewis(x) bound to LTL-A, or AAA revealed an unusual orientation of the fucose residue, with negative values for both dihedral angles, phi and psi, at the alpha(1-->3)-glycosidic linkage. A similar distortion of the fucose orientation was also observed for sialyl Lewis(x) bound to AAA. From STD NMR experiments it followed that only the L-fucose residues are in intimate contact with the protein. Presumably steric interactions are responsible for locking the sialic acid residue of sialyl Lewis(x) in one out of many orientations that are present in aqueous solution. The sialic acid residue of sialyl Lewis(x) bound to AAA adopts an orientation similar to that in the corresponding sialyl Lewis(x)/E-selectin complex.

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Section: Resultssupporting

confidence: 84%

Molecular Recognition of Sialyl Lewis^xand Related Saccharides by Two Lectins

2001

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“…Because both SPR and QCM techniques detect changes in mass upon binding, the small carbohydrate is usually immobilized and the large protein binding partner presented in solution. ,, Several examples of the reverse format (the addition of free sugar to immobilized lectin) have appeared, − but sensitivity and accuracy are generally limited . One attempt has been made to address this problem for carbohydrates with a heavy linker to enhance the mass-sensitive SPR signal upon binding …”

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confidence: 99%

Measurement of Monovalent and Polyvalent Carbohydrate−Lectin Binding by Back-Scattering Interferometry

et al. 2009

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Carbohydrate-protein binding is important to many areas of biochemistry. Back-scattering interferometry (BSI) is shown here to be a convenient and sensitive method for obtaining quantitative information about the strengths and selectivities of such interactions. The surfaces of glass microfluidic channels were covalently modified with extravidin, to which biotinylated lectins were subsequently attached by incubation and washing. The binding of unmodified carbohydrates to the resulting avidin-immobilized lectins was monitored by BSI. Dose-response curves, generated within several minutes and highly reproducible in multiple wash/measure cycles, provided adsorption coefficients that showed mannose to bind to concanavalin A with 3.7 times greater affinity than glucose, in line with literature values. Galactose was found to bind selectively and with similar affinity to the lectin BS-1. The avidities of polyvalent sugar-coated virus particles for immobilized conA were far higher than monovalent glycans, with increases of 60-200 fold per glycan when arrayed on the exterior surface of cowpea mosaic virus or bacteriophage Qβ. Sugar-functionalized PAMAM dendrimers showed size-dependent adsorption consistent with the expected density of lectins on the surface. The sensitivity of BSI matches or exceeds that of surface plasmon resonance and quartz crystal microbalance techniques, and differs in its sensitivity to the number of binding events rather than changes in mass. Its operational simplicity, generality, and the near-native conditions under which the target binding proteins are immobilized make it an attractive method for the quantitative characterization of the binding functions of lectins and other proteins.

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“…com). For convenience, these articles are subdivided into general biological categories: proteins, 45-183 antibodies, cell surface receptors, 283-337 peptides, small molecules, oligonucleotides, lipids and self-assembled monolayers, extracellular matrix, carbohydrates, [533][534][535][536][537][538][539][540][541][542] particles and viruses, [543][544][545] crude analytes, [546][547][548] and other configurations. [549][550][551][552][553][554] Forty-nine articles describe work performed using other optical biosensor instrumentation.…”

Section: Reviews Theory and Methodologymentioning

confidence: 99%

Survey of the year 2000 commercial optical biosensor literature

Rich

Myszka

2001

J of Molecular Recognition

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We have compiled a comprehensive list of the articles published in the year 2000 that describe work employing commercial optical biosensors. Selected reviews of interest for the general biosensor user are highlighted. Emerging applications in areas of drug discovery, clinical support, food and environment monitoring, and cell membrane biology are emphasized. In addition, the experimental design and data processing steps necessary to achieve high-quality biosensor data are described and examples of well-performed kinetic analysis are provided.

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Low Affinity Carbohydrate Lectin Interactions Examined with Surface Plasmon Resonance

Cited by 8 publications

References 24 publications

Molecular Recognition of Sialyl Lewis^xand Related Saccharides by Two Lectins

Molecular Recognition of Sialyl Lewis^xand Related Saccharides by Two Lectins

Measurement of Monovalent and Polyvalent Carbohydrate−Lectin Binding by Back-Scattering Interferometry

Survey of the year 2000 commercial optical biosensor literature

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Low Affinity Carbohydrate Lectin Interactions Examined with Surface Plasmon Resonance

Cited by 8 publications

References 24 publications

Molecular Recognition of Sialyl Lewisxand Related Saccharides by Two Lectins

Molecular Recognition of Sialyl Lewisxand Related Saccharides by Two Lectins

Measurement of Monovalent and Polyvalent Carbohydrate−Lectin Binding by Back-Scattering Interferometry

Survey of the year 2000 commercial optical biosensor literature

Contact Info

Product

Resources

About

Molecular Recognition of Sialyl Lewis^xand Related Saccharides by Two Lectins

Molecular Recognition of Sialyl Lewis^xand Related Saccharides by Two Lectins