Mutational analysis of ProTx-I and the novel venom peptide Pe1b provide insight into residues responsible for selective inhibition of the analgesic drug target NaV1.7

Rupasinghe, Darshani B.; Herzig, Volker; Vetter, Irina; Dekan, Zoltan; Bosmans, Frank; Alewood, Paul F.; Lewis, Richard J.; King, Glenn F.

doi:10.1016/j.bcp.2020.114080

Cited by 10 publications

(7 citation statements)

References 51 publications

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“…As part of our ongoing attempts to develop new pharmacological probes and therapeutic leads for human Na V channels, ,− we devised and tested in this study a bivalent linker design with a focus on Na V 1.4 and Na V 1.7 due to their (patho)physiological relevance and experimentally determined structures. Our strategy was to covalently link a pore blocker toxin with a gating modifier toxin using variable-length polyethylene glycol (PEG) linkers to simultaneously target two binding sites of the channel, thereby potentially enhancing binding kinetics, potency, and subtype selectivity (Figure B,C).…”

Section: Introductionmentioning

confidence: 99%

Two for the Price of One: Heterobivalent Ligand Design Targeting Two Binding Sites on Voltage-Gated Sodium Channels Slows Ligand Dissociation and Enhances Potency

et al. 2020

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Voltage-gated sodium (Na V ) channels are pore-forming transmembrane proteins that play essential roles in excitable cells, and they are key targets for antiepileptic, antiarrhythmic, and analgesic drugs. We implemented a heterobivalent design strategy to modulate the potency, selectivity, and binding kinetics of Na V channel ligands. We conjugated μ-conotoxin KIIIA, which occludes the pore of the Na V channels, to an analogue of huwentoxin-IV, a spider-venom peptide that allosterically modulates channel gating. Bioorthogonal hydrazide and copper-assisted azide–alkyne cycloaddition conjugation chemistries were employed to generate heterobivalent ligands using polyethylene glycol linkers spanning 40–120 Å. The ligand with an 80 Å linker had the most pronounced bivalent effects, with a significantly slower dissociation rate and 4–24-fold higher potency compared to those of the monovalent peptides for the human Na V 1.4 channel. This study highlights the power of heterobivalent ligand design and expands the repertoire of pharmacological probes for exploring the function of Na V channels.

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Section: Introductionmentioning

confidence: 99%

Two for the Price of One: Heterobivalent Ligand Design Targeting Two Binding Sites on Voltage-Gated Sodium Channels Slows Ligand Dissociation and Enhances Potency

et al. 2020

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“…This suggests that this pH-dependence could potentially be exploited in the design of peptides as a “molecular switch” for use in the gastrointestinal tract, , lysosomes, secretory granules, and pH-altering states such as cancer, − inflammation, , ischemia, and others . Peptides containing acidic residues that are protonated in acidic environments to promote membrane insertion have been studied for cancer and inflammation, − , and other pH-sensitive peptides have been studied for various applications. − …”

Section: Discussionmentioning

confidence: 99%

Structural Conformation and Activity of Spider-Derived Inhibitory Cystine Knot Peptide Pn3a Are Modulated by pH

et al. 2023

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Numerous spider venom-derived gating modifier toxins exhibit conformational heterogeneity during purification by reversed-phase high-performance liquid chromatography (RP-HPLC). This conformational exchange is especially peculiar for peptides containing an inhibitor cystine knot motif, which confers excellent structural stability under conditions that are not conducive to disulfide shuffling. This phenomenon is often attributed to proline cis/trans isomerization but has also been observed in peptides that do not contain a proline residue. Pn3a is one such peptide forming two chromatographically distinguishable peaks that readily interconvert following the purification of either conformer. The nature of this exchange was previously uncharacterized due to the fast rate of conversion in solution, making isolation of the conformers impossible. In the present study, an N-terminal modification of Pn3a enabled the isolation of the individual conformers, allowing activity assays to be conducted on the individual conformers using electrophysiology. The conformers were analyzed separately by nuclear magnetic resonance spectroscopy (NMR) to study their structural differences. RP-HPLC and NMR were used to study the mechanism of exchange. The later-eluting conformer was the active conformer with a rigid structure that corresponds to the published structure of Pn3a, while NMR analysis revealed the earliereluting conformer to be inactive and disordered. The exchange was found to be pH-dependent, arising in acidic solutions, possibly due to reversible disruption and formation of intramolecular salt bridges. This study reveals the nature of non-proline conformational exchange observed in Pn3a and possibly other disulfide-rich peptides, highlighting that the structure and activity of some disulfidestabilized peptides can be dramatically susceptible to disruption.

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“…The two peptides (34 residues) β/μ-TRTX-Pe1a and β/μ-TRTX-Pe1b from the arboreal tarantula Phormingochilus everetti are Nav1.7 inhibitors. The recombinant Pe1b peptide shows potent and relative selective inhibition (IC 50 167nM) on Nav1.7 compared to other Nav isoforms such as Nav1.6, Nav1.5 and Nav1.2 [ 199 ].…”

Section: Spider Toxins Interacting With Pain-related Ion Channelsmentioning

confidence: 99%

“…ProTx analogues were designed to improve selectivity for Nav1.7 based on peptide pharmacophore and channel interaction [ 199 ]. Several mutants, with modified C terminus (ProTx-I-NH2 and ProTx-II-NHCH3), have increased activity on both Nav1.2 and Nav1.7 and a decreased activity on Nav1.5 and Nav1.6 [ 240 ].…”

Section: Spider Toxins Interacting With Pain-related Ion Channelsmentioning

confidence: 99%

Pain-related toxins in scorpion and spider venoms: a face to face with ion channels

Diochot

2021

J. Venom. Anim. Toxins incl. Trop. Dis

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Pain is a common symptom induced during envenomation by spiders and scorpions. Toxins isolated from their venom have become essential tools for studying the functioning and physiopathological role of ion channels, as they modulate their activity. In particular, toxins that induce pain relief effects can serve as a molecular basis for the development of future analgesics in humans. This review provides a summary of the different scorpion and spider toxins that directly interact with pain-related ion channels, with inhibitory or stimulatory effects. Some of these toxins were shown to affect pain modalities in different animal models providing information on the role played by these channels in the pain process. The close interaction of certain gating-modifier toxins with membrane phospholipids close to ion channels is examined along with molecular approaches to improve selectivity, affinity or bioavailability in vivo for therapeutic purposes.

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Mutational analysis of ProTx-I and the novel venom peptide Pe1b provide insight into residues responsible for selective inhibition of the analgesic drug target NaV1.7

Cited by 10 publications

References 51 publications

Two for the Price of One: Heterobivalent Ligand Design Targeting Two Binding Sites on Voltage-Gated Sodium Channels Slows Ligand Dissociation and Enhances Potency

Two for the Price of One: Heterobivalent Ligand Design Targeting Two Binding Sites on Voltage-Gated Sodium Channels Slows Ligand Dissociation and Enhances Potency

Structural Conformation and Activity of Spider-Derived Inhibitory Cystine Knot Peptide Pn3a Are Modulated by pH

Pain-related toxins in scorpion and spider venoms: a face to face with ion channels

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