Thermodynamic Studies of Lectin−Carbohydrate Interactions by Isothermal Titration Calorimetry

Dam, Tarun K.; Brewer, C. Fred

doi:10.1021/cr000401x

Cited by 445 publications

(381 citation statements)

References 213 publications

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“…These results indicated that the saccharide-immobilized substrates have strong and specific interaction with the saccharide recognition proteins of lectins. The binding constants (K A ) of the monovalent saccharide-protein in solution were much stronger in our experiment than in [21]. The strong binding affinities were contributed by the multivalent effect, because the immobilized saccharide formed a densely packed monolayer [3,4].…”

Section: Quantitative Analyses Of Lectin Recognition On the Microarraymentioning

confidence: 57%

Sugar microarray via click chemistry: molecular recognition with lectins and amyloid β (1–42)

Matsumoto

Yamauchi

Fukuda

et al. 2009

Science and Technology of Advanced Materials

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Sugar microarrays were fabricated on various substrates via click chemistry. Acetylene-terminated substrates were prepared by forming self-assembled monolayers (SAMs) on a gold substrate with alkyl-disulfide and on silicon, quartz and glass substrates with a silane-coupling reagent. The gold substrates were subjected to surface plasmon resonance measurements, and the quartz and glass substrates were subjected to spectroscopy measurements and optical microscopy observation. The saccharide-immobilized substrate on the gold substrate showed specific interaction with the corresponding lectin, and the saccharides showed inert surface properties to other proteins with a high signal-to-noise ratio. We also focused on the saccharide-protein interaction on protein amyloidosis of Alzheimer amyloid β. Amyloid β peptide showed conformation transition on the saccharide-immobilization substrate into a β-sheet, and fibril formation and amyloid aggregates were found on the specific saccharides.

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Section: Quantitative Analyses Of Lectin Recognition On the Microarraymentioning

confidence: 57%

Sugar microarray via click chemistry: molecular recognition with lectins and amyloid β (1–42)

Matsumoto

Yamauchi

Fukuda

et al. 2009

Science and Technology of Advanced Materials

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“…Such lowaffinity, high-specificity recognition systems are gaining increasing importance in serving key biological functions, including T cell receptor recognition of peptide͞MHC ligands (36)(37)(38), cell surface carbohydrate-protein interactions (39)(40)(41), and cell-cell adhesion (42). Moreover, it may be that even seemingly small (Ͻ10-fold) differences in the affinity of PGRPs for monovalent PGN ligands, such as we measured in several cases, are amplified by multiple PGRP-PGN interactions to establish specificity effects at the cellular level, as described for carbohydratebinding proteins (43,44).…”

Section: Discussionmentioning

confidence: 99%

Dual strategies for peptidoglycan discrimination by peptidoglycan recognition proteins (PGRPs)

Swaminathan

Brown

Roychowdhury

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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The innate immune system constitutes the first line of defense against microorganisms in both vertebrates and invertebrates. Although much progress has been made toward identifying key receptors and understanding their role in host defense, far less is known about how these receptors recognize microbial ligands. Such studies have been severely hampered by the need to purify ligands from microbial sources and a reliance on biological assays, rather than direct binding, to monitor recognition. We used synthetic peptidoglycan (PGN) derivatives, combined with microcalorimetry, to define the binding specificities of human and insect peptidogycan recognition proteins (PGRPs). We demonstrate that these innate immune receptors use dual strategies to distinguish between PGNs from different bacteria: one based on the composition of the PGN peptide stem and another that senses the peptide bridge crosslinking the stems. To pinpoint the site of PGRPs that mediates discrimination, we engineered structure-based variants having altered PGN-binding properties. The plasticity of the PGRPbinding site revealed by these mutants suggests an intrinsic capacity of the innate immune system to rapidly evolve specificities to meet new microbial challenges.affinity ͉ bacteria ͉ innate immunity ͉ calorimetry ͉ synthesis T he innate immune system recognizes invading microbes by means of conserved pattern recognition receptors that bind unique products of microbial metabolism not produced by the host (pathogen-associated molecular patterns) (1, 2). Examples of microbial ligands recognized by pattern recognition receptors such as Toll-like receptors, peptidoglycan recognition proteins (PGRPs), and NOD proteins include lipopolysaccharide of Gram-negative bacteria, lipoteichoic acid of Gram-positive bacteria, nonmethylated CpG sequences, flagellin, and peptidoglycan (PGN) of Gram-negative and -positive bacteria. Cellular activation by pattern recognition receptors results in acute inflammatory responses involving cytokine and chemokine production, direct local attack against the invading pathogen, and induction of the adaptive component of the immune system. In humans, overactivation of inflammatory responses can lead to septic shock, which accounts for 100,000 deaths annually in the United States alone. By sitting at the intersection of the pathways of microbial recognition, inflammation, and cell death, the innate immune system offers emerging opportunities for the development of therapeutics to modulate immune responses (3).PGRPs, a newly discovered class of pattern recognition receptors, are highly conserved from insects to mammals (4-7). By detecting PGN from both Gram-negative and -positive bacteria, PGRPs are important contributors to host defense against microbial infections (2, 4). PGNs are polymers of alternating Nacetylglucosamine (GlcNAc) and N-acetylmuramic acid (MurNAc) in ␤(134) linkage, crosslinked by short peptide stems composed of alternating L-and D-amino acids (8, 9) (Fig. 1A). Whereas the carbohydrate backbone is conserved a...

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“…Schematic and CFG representation of compounds 1 -5 studied using SEAL by NMR. Models of the lectins were generated by the PyMol software (The PyMOL Molecular Graphics System, Version 1.5.0.1 Schrödinger, LLC) and originate from X-ray crystal structures (WGA [30] : PDB ID: 2UVO; ConA [32] : PDB ID: 5CNA; PNA [33] : PDB ID: 2PEL). The ligands are represented as 3D-CFG.…”

mentioning

confidence: 99%

“…As indicated in Figure 1, WGA binds selectively to N-acetyl-D-glucosamine, ConA to α-Dmannose and PNA to D-galactose and the affinities (K D ) are in the range of mM to µM according to literature data. [30] Our approach is to study combinations of the selenoglycosides in the absence (blank) and presence of protein, employing otherwise identical conditions, and directly monitoring different parameters that change upon complexation by regular proton decoupled 1D 77 Se NMR spectra, see Figure 2. In order to display the effects of ligand-binding unbiased from medium effects, all the experiments were carried out with methyl 1-seleno-β-Dglucopyranoside (GlcSeMe, 1) as a non-interfering reference compound present in an NMR insert-tube.…”

mentioning

confidence: 99%

“…[30] However, comparing x-ray crystal structures of the natural ligands for the lectins studied [31][32][33] (see Figure 1), it is clear that only in the case of WGA the anomeric oxygen (position for selenium substitution) is in close proximity to a protein amino acid residue in the binding pocket. This is revealed by the fact that the chemical shift differences between the GNAcSeMe in presence and absence of WGA is the largest seen in the study (~17 Hz) indicating an altered chemical environment between free and bound state.…”

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confidence: 99%

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SEAL by NMR: Glyco‐Based Selenium‐Labeled Affinity Ligands Detected by NMR Spectroscopy

Hamark

Landström

Widmalm

2014

Chemistry A European J

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Abstract:We report a method for the screening of interactions between proteins and selenium-labeled carbohydrate ligands. SEAL by NMR is demonstrated with selenoglycosides binding to lectins where the selenium nucleus serves as an NMR-active handle and reports on binding through 77 Se NMR spectroscopy. In terms of overall sensitivity, this nucleus is comparable to 13 C NMR while the NMR spectral width is ten times larger, yielding little overlap in 77 Se NMR spectroscopy, even for similar compounds. The studied ligands are singly-selenated bioisosteres of methyl glycosides for which straightforward preparation methods are at hand and libraries can readily be generated. The strength of the approach lies in its simplicity, sensitivity to binding events, the tolerance to additives and the possibility of having several ligands in the assay. This study extends the increasing potential of selenium in structure biology and medicinal chemistry. We anticipate that SEAL by NMR will be a beneficial tool for the development of selenium-based bioactive compounds, such as glycomimetic drug candidates.

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Thermodynamic Studies of Lectin−Carbohydrate Interactions by Isothermal Titration Calorimetry

Cited by 445 publications

References 213 publications

Sugar microarray via click chemistry: molecular recognition with lectins and amyloid β (1–42)

Sugar microarray via click chemistry: molecular recognition with lectins and amyloid β (1–42)

Dual strategies for peptidoglycan discrimination by peptidoglycan recognition proteins (PGRPs)

SEAL by NMR: Glyco‐Based Selenium‐Labeled Affinity Ligands Detected by NMR Spectroscopy

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