Probing pattern and dynamics of disulfide bridges using synthesis and NMR of an ion channel blocker peptide toxin with multiple diselenide bonds

Fehér, Krisztina; Timári, István; Rákosi, Kinga; Szolomájer, János; Illyés, Tünde Zita; Bartók, Ádám; Varga, Zoltán; Panyi, György; Tóth, Gábor; Kövér, Katalin E.

doi:10.1039/c5sc03995a

Cited by 7 publications

(8 citation statements)

References 47 publications

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“…Biomolecular simulations have become a method of choice to gain a detailed insight into the structure and dynamic properties of protein–ligand complexes [ 44 , 45 , 46 , 47 , 48 ]. To gain insight into the energetics of the selenium disaccharides binding to h Gal-3 CRD, we carried out multiple MD simulations for 1000 ns.…”

Section: Resultsmentioning

confidence: 99%

Investigation of the Molecular Details of the Interactions of Selenoglycosides and Human Galectin-3

Raics

Balogh

Kishor

et al. 2022

IJMS

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Human galectin-3 (hGal-3) is involved in a variety of biological processes and is implicated in wide range of diseases. As a result, targeting hGal-3 for clinical applications has become an intense area of research. As a step towards the development of novel hGal-3 inhibitors, we describe a study of the binding of two Se-containing hGal-3 inhibitors, specifically that of di(β-D-galactopyranosyl)selenide (SeDG), in which two galactose rings are linked by one Se atom and a di(β-D-galactopyranosyl)diselenide (DSeDG) analogue with a diseleno bond between the two sugar units. The binding affinities of these derivatives to hGal-3 were determined by 15N-1H HSQC NMR spectroscopy and fluorescence anisotropy titrations in solution, indicating a slight decrease in the strength of interaction for SeDG compared to thiodigalactoside (TDG), a well-known inhibitor of hGal-3, while DSeDG displayed a much weaker interaction strength. NMR and FA measurements showed that both seleno derivatives bind to the canonical S face site of hGal-3 and stack against the conserved W181 residue also confirmed by X-ray crystallography, revealing canonical properties of the interaction. The interaction with DSeDG revealed two distinct binding modes in the crystal structure which are in fast exchange on the NMR time scale in solution, explaining a weaker interaction with hGal-3 than SeDG. Using molecular dynamics simulations, we have found that energetic contributions to the binding enthalpies mainly differ in the electrostatic interactions and in polar solvation terms and are responsible for weaker binding of DSeDG compared to SeDG. Selenium-containing carbohydrate inhibitors of hGal-3 showing canonical binding modes offer the potential of becoming novel hydrolytically stable scaffolds for a new class of hGal-3 inhibitors.

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Section: Resultsmentioning

confidence: 99%

Investigation of the Molecular Details of the Interactions of Selenoglycosides and Human Galectin-3

Raics

Balogh

Kishor

et al. 2022

IJMS

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“…CPMG-INEPT long-range transfer eliminates signal phase modulation and significantly reduces intensity losses from co-evolving J( 1 H, 1 H) couplings. Furthermore, it can suppress line broadening from chemical exchange, ensuring maximum detection sensitivity and clean signal phases, as was described earlier [25,26,32].…”

Section: Nmr Experimentsmentioning

confidence: 99%

“…To improve sensitivity, we proposed a 2D 1 H-77 Se HSQMBC (heteronuclear single quantum multiple-bond correlation) experiment via indirect 77 Se detection using CPMG-INEPT (Carr-Purcell-Meiboom-Gill insensitive nuclei enhanced by polarization transfer) out-and-back 1 H→ 77 Se→ 1 H polarization transfer. Theoretically, this approach can yield up to 60-fold ((γ H /γ Se ) 2.5 ) sensitivity enhancement, but, owing to competing relaxation and other transfer processes, a sensitivity gain about 20 can be realized in practice [25,28,32].…”

Section: Introductionmentioning

confidence: 99%

77Se-Enriched Selenoglycoside Enables Significant Enhancement in NMR Spectroscopic Monitoring of Glycan–Protein Interactions

et al. 2022

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Detailed investigation of ligand–protein interactions is essential for better understanding of biological processes at the molecular level. Among these binding interactions, the recognition of glycans by lectins is of particular importance in several diseases, such as cancer; therefore, inhibition of glycan-lectin/galectin interactions represents a promising perspective towards developing therapeutics controlling cancer development. The recent introduction of 77Se NMR spectroscopy for monitoring the binding of a selenoglycoside to galectins prompted interest to optimize the sensitivity by increasing the 77Se content from the natural 7.63% abundance to 99%. Here, we report a convenient synthesis of 77Se-enriched selenodigalactoside (SeDG), which is a potent ligand of the medically relevant human galectin-3 protein, and proof of the expected sensitivity gain in 2D 1H, 77Se correlation NMR experiments. Our work opens perspectives for adding isotopically enriched selenoglycans for rapid monitoring of lectin-binding of selenated as well as non-selenated ligands and for ligand screening in competition experiments.

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“…[17] The high tolerance of diselenide bondst or educing conditions was successfully exploited in severalc ases to deliver fully active peptidem imetics where the native 3D structure was preserved and disulfide scrambling effectively prevented. [18][19][20][21][22] However, since peptides degradation in plasma largely occurs by endo-a nd exo-peptidases rather than oxido-reductases, enhanced reductive stabilityn ot always results in am arkedlyi mprovedm etabolic profile. Ac ase example of successfuld isulfide-to-diselenide application was recently reported for the synthesis of the seleno analogue of bovine pancreatic insulin by Iwaoka and collaborators.…”

Section: Diselenide Bond Analoguesmentioning

confidence: 99%

“…The high tolerance of diselenide bonds to reducing conditions was successfully exploited in several cases to deliver fully active peptide mimetics where the native 3D structure was preserved and disulfide scrambling effectively prevented . However, since peptides degradation in plasma largely occurs by endo ‐ and exo ‐peptidases rather than oxido‐reductases, enhanced reductive stability not always results in a markedly improved metabolic profile.…”

Section: Diselenide Bond Analoguesmentioning

confidence: 99%

Disulfide Bond Mimetics: Strategies and Challenges

Gori

Gagni

Rinaldi

2017

Chemistry A European J

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The activity profile of many biologically relevant proteins and peptides often relies on a precise 3D structural organization. In this context, disulfide bonds are natural covalent constraints that play a key role in driving and stabilizing the folding pattern of these molecules. Despite its prominent significance as structural motif, the disulfide bond itself is inherently unstable under physiological conditions, posing a major limit to the use and development of disulfide-rich peptides and proteins as molecular tools and drug lead compounds. To tackle this restriction, disulfide engineering with stable functional analogues has arisen a considerable interest. Here, the most popular approaches to disulfide replacement are reviewed and discussed with particular emphasis on advantages and limitations under both functional and synthetic perspectives.

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Probing pattern and dynamics of disulfide bridges using synthesis and NMR of an ion channel blocker peptide toxin with multiple diselenide bonds

Abstract: A biologically active peptide toxin containing four diselenide bonds was synthesized. The diselenide network and its dynamics were disclosed using a combined NMR and MD approach.

Cited by 7 publications

References 47 publications

Investigation of the Molecular Details of the Interactions of Selenoglycosides and Human Galectin-3

Investigation of the Molecular Details of the Interactions of Selenoglycosides and Human Galectin-3

77Se-Enriched Selenoglycoside Enables Significant Enhancement in NMR Spectroscopic Monitoring of Glycan–Protein Interactions

Disulfide Bond Mimetics: Strategies and Challenges

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