Cryo-EM structure of the ASIC1a–mambalgin-1 complex reveals that the peptide toxin mambalgin-1 inhibits acid-sensing ion channels through an unusual allosteric effect

Sun, Degang; Yu, You; Xue, Xiaobin; Pan, Man; Wen, Ming; Li, Siyu; Qu, Qian; Li, Xiaorun; Zhang, Longhua; Li, Xueming; Liu, Lei; Yang, Maojun; Tian, Changlin

doi:10.1038/s41421-018-0026-1

Cited by 36 publications

(53 citation statements)

References 55 publications

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“…; Sun et al . ). The swallowing threshold induced by a series of von Frey filaments was measured before and after topical administration of 3 μl aliquots of amiloride (0.3–30 nmol), benzamil (0.3–30 nmol), dimethylamiloride (0.3–30 nmol), DMSO (1%–100%), gadolinium (0.3–30 nmol), mambalgin‐1 (0.3–30 nmol) and diminazene (0.3–30 nmol).…”

Section: Methodsmentioning

confidence: 97%

“…We used the ENaC blocker amiloride, its analogues benzamil and dimethylamiloride, the vehicle of amiloride analogues dimethyl sulfoxide (DMSO), the classic mechanosensitive channel blocker gadolinium, and the ASIC inhibitors mambalgine-1 and diminazene. Mambalgine-1 is an ASIC-1a and -1b inhibitor, and diminazene is an ASIC-1a, -1b, -2a and -3 inhibitor (Chen et al 2010;Diochot et al 2012;Sun et al 2018). The swallowing threshold induced by a series of von Frey filaments was measured before…”

Section: Effect Of Nerve Transection and Topical Drug Application On mentioning

confidence: 99%

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Involvement of the epithelial sodium channel in initiation of mechanically evoked swallows in anaesthetized rats

Tsujimura

Ueha

Yoshihara

et al. 2019

The Journal of Physiology

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Key points Afferents carried by the superior laryngeal nerve play a primary role in the initiation of laryngeal mechanically evoked swallows in anaesthetized rats. Amiloride and its analogues inhibit swallowing evoked by mechanical stimulation, but not swallowing evoked by chemical and electrical stimulation. The epithelial sodium channel is probably involved in the initiation of laryngeal mechanically evoked swallows. Abstract The swallowing reflex plays a critical role in airway protection. Because impaired laryngeal mechanosensation is associated with food bolus aspiration, it is important to know how the laryngeal sensory system regulates swallowing initiation. This study was performed to clarify the neuronal mechanism of mechanically evoked swallows. Urethane‐anaesthetized Sprague–Dawley male rats were used. A swallow was identified by activation of the suprahyoid and thyrohyoid muscles on electromyography. The swallowing threshold was measured by von Frey filament and electrical stimulation of the larynx. The number of swallows induced by upper airway distension and capsaicin application (0.03 nmol, 3 μl) to the vocal folds was counted. The effects of topical application (0.3–30 nmol, 3 μl) of the epithelial sodium channel (ENaC) blocker amiloride and its analogues (benzamil and dimethylamiloride), acid‐sensing ion channel (ASIC) inhibitors (mambalgine‐1 and diminazene) and gadolinium to the laryngeal mucosa on swallowing initiation were evaluated. A nerve transection study indicated that afferents carried by the superior laryngeal nerve play a primary role in the initiation of laryngeal mechanically evoked swallows. The mechanical threshold of swallowing was increased in a dose‐dependent manner by amiloride and its analogues and gadolinium, but not by ASIC inhibitors. The number of swallows by upper airway distension was significantly decreased by benzamil application. However, the initiation of swallows evoked by capsaicin and electrical stimulation was not affected by benzamil application. We speculate that the ENaC is involved in the initiation of laryngeal mechanically evoked swallows.

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Section: Methodsmentioning

confidence: 97%

Section: Effect Of Nerve Transection and Topical Drug Application On mentioning

confidence: 99%

Involvement of the epithelial sodium channel in initiation of mechanically evoked swallows in anaesthetized rats

Tsujimura

Ueha

Yoshihara

et al. 2019

The Journal of Physiology

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“…Mourier et al performed full concentration-response curves, and showed >10fold loss in IC50 for mutants F27A, R28A, L32A, I33A, and L34A, concluding that these residues are crucial for the ASIC1a interaction. A second study used cASIC1a and tested the inhibitory activity of several Ma-1 mutants (Q5A, H6A, K8A, F27A, and R28A) at a single concentration of 500 nM, extending the peptide mutagenesis to include finger I (89). Using a single concentration of Ma-1, it is difficult to determine the true extent of loss in activity of each mutant peptide.…”

Section: Discussionmentioning

confidence: 99%

“…The binding site of mambalgins on ASICs has been studied by several groups using a combination of functional and structural techniques as well as docking studies. However, there is disagreement between these studies over the proposed binding orientation of mambalgins at ASICs (see Figure 6.11 for published models) (44,45,50,89,157). produced by rigid body fitting the cASIC1a desensitised crystal structure (PDB code 4FZ1) extracellular domain and mambalgin-1 crystal structure (PDB code 5DU1) into the cryo-EM density,…”

Section: Discussionmentioning

confidence: 99%

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Discovery and modulation of acid-sensing ion channel modulating venom peptides

Cristofori-Armstrong¹

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Acid-sensing ion channels (ASICs) are a family of proton-activated cation channels expressed in a variety of neuronal and non-neuronal tissues. As proton-gated channels, they have been implicated in many pathophysiological conditions where pH is perturbed. Venom derived compounds represent the most potent and selective modulators of ASICs described to date, and thus have been invaluable pharmacological tools to study ASIC structure, function, and biological roles. There are now eleven ASIC modulators described from animal venoms, with those from snakes and spiders favouring ASIC1, while the sea anemone and cone snail modulators preferentially target ASIC3. Chapter 1 reviews the current state of knowledge on venom derived ASIC modulators, with a particular focus on their molecular interaction with ASICs, what they have taught us about channel structure, and what they may still reveal about ASIC function and pathophysiological roles. Venom peptides are often disulfide bonded making their recombinant expression challenging. Chapter 2 describes a periplasmic Escherichia coli protocol for production of correctly folded peptides that was used throughout this thesis. Using the two-electrode voltage-clamp technique, Chapter 3 shows that a wide variety of voltage-and ligand-gated ion channels have the same channel properties and pharmacological profiles when expressed in either Xenopus laevis or X. borealis oocytes. This voltage-clamp technique was heavily used throughout this thesis to perform functional studies of venom peptides. The spider venom peptide PcTx1 is the best studied ASIC modulator; it has an IC50 of ~1 nM at rat ASIC1a and is neuroprotective in rodent models of ischemic stroke. Chapter 4 examines the molecular interaction between PcTx1 and both human ASIC1a and the offtarget ASIC1b subtype, where little experimental work has been done. We show that although PcTx1 is 10-fold less potent at human ASIC1a than the rat channel, the apparent affinity for the two channels is comparable. The pharmacophore of PcTx1 for human ASIC1a and rat ASIC1b was examined via alanine scanning mutagenesis and uncovered residues that show subtle ASIC1 species and subtype-dependent differences in activity that may allow for further manipulation to develop more selective PcTx1 analogues. The ASIC3 inhibitor APETx2 is analgesic in rodent models of chemically-induced, inflammatory and postoperative pain, providing strong evidence for the ASIC3 subtype as a potential pain target. Despite the comprehensive structure-activity studies of APETx2, the mechanism of action and channel binding site have remained elusive. Chapter 5 fills this VII partial and highly variable antihyperalgesia in a rat model of inflammatory pain. British

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A Diaminodiacid (DADA) Strategy for the Development of Disulfide Surrogate Peptides

Bierer

et al. 2020

Chemistry An Asian Journal

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Disulfide bond‐containing peptides are useful molecular scaffolds with diagnostic and therapeutic applications due to their good biological activity and good target selectivity, but their utility is sometimes limited by the lability of the disulfide moiety under reducing conditions and in the presence of disulfide bond isomerase. The development of disulfide surrogates with improved redox stability has been an area of ongoing research; and one possible strategy is based on a diaminodiacid (DADA) moiety, which can be used to synthesize the disulfide bond replacement peptides with precise structures and enhanced stability through automated solid‐phase peptide synthesis (SPPS). This review summarizes recent developments in the DADA‐based SPPS of peptide disulfide surrogates. Some representative applications and structural studies on the DADA‐based disulfide surrogates are described.

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Cryo-EM structure of the ASIC1a–mambalgin-1 complex reveals that the peptide toxin mambalgin-1 inhibits acid-sensing ion channels through an unusual allosteric effect

Cited by 36 publications

References 55 publications

Involvement of the epithelial sodium channel in initiation of mechanically evoked swallows in anaesthetized rats

Involvement of the epithelial sodium channel in initiation of mechanically evoked swallows in anaesthetized rats

Discovery and modulation of acid-sensing ion channel modulating venom peptides

A Diaminodiacid (DADA) Strategy for the Development of Disulfide Surrogate Peptides

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