<scp>NMR</scp> structure and dynamics of the <scp>C</scp>‐terminal domain of <scp>R</scp>‐type lectin from the earthworm <i><scp>L</scp>umbricus terrestris</i>

Hemmi, Hikaru; Kuno, Atsushi; Hirabayashi, Jun

doi:10.1111/febs.12050

Cited by 8 publications

(1 citation statement)

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“…We focused here on NMR relaxation measurements as these provide direct insights into local motions at defined atomic sites in molecules. As shown in recent examples, − such NMR relaxation data can facilitate an understanding of molecular binding properties and mechanisms of action.…”

Section: Introductionmentioning

confidence: 94%

Insights into the Molecular Flexibility of θ-Defensins by NMR Relaxation Analysis

Conibear¹,

Wang²,

et al. 2014

J. Phys. Chem. B

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θ-Defensins are mammalian cyclic peptides that have antimicrobial activity and show potential as stable scaffolds for peptide-based drug design. The cyclic cystine ladder structural motif of θ-defensins has been characterized using NMR spectroscopy and is important for their structure and stability. However, the effect of the pronounced elongated topology of θ-defensins on their molecular motion is not yet understood. Studies of molecular motion by NMR relaxation measurements have been facilitated by the recent development of a semirecombinant method for producing cyclic peptides that allows for isotopic labeling. Here we have undertaken a multifield 15N NMR relaxation analysis of the anti-HIV θ-defensin, HTD-2, and interpreted the experimental data using various models of overall and internal molecular motion. We found that it was necessary to apply a model that includes internal motion to account for the variations in the experimental T1 and NOE data at different backbone amide sites in the peptide. Although an isotropic model with internal motion was the simplest model that provided a satisfactory fit with the experimental data, we cannot exclude the possibility that overall motion is anisotropic, especially considering the strikingly elongated topology of θ-defensins. The presence of flexible side chains, self-association, interactions with solvent, and internal motions are all potential contributors to the observed relaxation data. Internal motion consistent with the constraints imposed by the cyclic cystine ladder was observed in that the order parameters, S2, show that residues in the turns are more flexible than those in the β-sheet. This study provides insights into the dynamics of θ-defensins and information that might be useful in their application as scaffolds in drug design.

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Section: Introductionmentioning

confidence: 94%

Insights into the Molecular Flexibility of θ-Defensins by NMR Relaxation Analysis

Conibear¹,

Wang²,

et al. 2014

J. Phys. Chem. B

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NMR analysis on the sialic acid‐binding mechanism of an R‐type lectin mutant by natural evolution‐mimicry

et al. 2016

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A sialic acid-binding lectin (SRC) was created from the C-terminal domain of an R-type N-acetyl lactosamine-binding lectin (EW29Ch) by natural evolution-mimicry. Here, we clarified its sialic acid-binding mechanism using NMR spectroscopy. The NMR analysis showed differences between conformations of the 6 0 -sialyllactose-bound SRC in the solution state and that in the crystal state, and differences between the internal motion of the loop region in subdomain c in SRC and that of the corresponding region in EW29Ch. The NMR analysis thus provided useful information to explain the manner of binding to 6 0 -sialyllactose in solution, which the previous X-ray crystal structure analysis lacked.

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Coagulant Activity of Water-Soluble Moringa oleifera Lectin Is Linked to Lowering of Electrical Resistance and Inhibited by Monosaccharides and Magnesium Ions

Moura

Silva

Dornelles

et al. 2016

Appl Biochem Biotechnol

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Moringa oleifera seeds contain a water-soluble lectin [water-soluble M. oleifera lectin (WSMoL)] that has shown coagulant activity. Magnesium ions are able to interfere with the ability of this lectin to bind carbohydrates. In this study, we performed structural characterization of WSMoL and analyzed its effect on the electrical resistance of a kaolin clay suspension in both presence and absence of monosaccharides (N-acetylglucosamine, glucose, or fructose) and magnesium ions. The coagulant activity of WSMoL was monitored by measuring optical density and electrical resistance over a period of 60 min. Native WSMoL had a molecular mass of 60 kDa and exhibited anionic nature (pI 5.5). In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), it appeared as three polypeptide bands of 30, 20, and 10 kDa. WSMoL reduced the optical density and electrical resistance of the kaolin suspension, which suggests that suspended particles are destabilized and that this is followed by formation of complexes. The coagulant activity of lectin decreased in the presence of Mg ions and carbohydrates at concentrations that also inhibited hemagglutinating activity. This was most likely due to conformational changes in lectin structure. Our findings suggest that the coagulant activity of WSMoL is enhanced by lowering of electrical resistance of the medium and is impaired by lectin-carbohydrate and lectin-Mg interactions.

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NMR structure and dynamics of the C‐terminal domain of R‐type lectin from the earthworm Lumbricus terrestris

Cited by 8 publications

References 50 publications

Insights into the Molecular Flexibility of θ-Defensins by NMR Relaxation Analysis

Insights into the Molecular Flexibility of θ-Defensins by NMR Relaxation Analysis

NMR analysis on the sialic acid‐binding mechanism of an R‐type lectin mutant by natural evolution‐mimicry

Coagulant Activity of Water-Soluble Moringa oleifera Lectin Is Linked to Lowering of Electrical Resistance and Inhibited by Monosaccharides and Magnesium Ions

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