scholarly journals Resting state structure of the hyperdepolarization activated two-pore channel 3

2020 ◽  
Vol 117 (4) ◽  
pp. 1988-1993
Author(s):  
Miles Sasha Dickinson ◽  
Alexander Myasnikov ◽  
Jacob Eriksen ◽  
Nicole Poweleit ◽  
Robert M. Stroud
Keyword(s):  
Resting State ◽  
Action Potentials ◽  
Pore Channel ◽  
Activation Threshold ◽  
Voltage Gated ◽  
Bona Fide ◽  
Voltage Gated Ion Channels ◽  
Voltage Sensing Domain ◽  
Voltage Activation ◽  
Closed Pore

Voltage-gated ion channels endow membranes with excitability and the means to propagate action potentials that form the basis of all neuronal signaling. We determined the structure of a voltage-gated sodium channel, two-pore channel 3 (TPC3), which generates ultralong action potentials. TPC3 is distinguished by activation only at extreme membrane depolarization (V50 ∼ +75 mV), in contrast to other TPCs and NaV channels that activate between −20 and 0 mV. We present electrophysiological evidence that TPC3 voltage activation depends only on voltage sensing domain 2 (VSD2) and that each of the three gating arginines in VSD2 reduces the activation threshold. The structure presents a chemical basis for sodium selectivity, and a constricted gate suggests a closed pore consistent with extreme voltage dependence. The structure, confirmed by our electrophysiology, illustrates the configuration of a bona fide resting state voltage sensor, observed without the need for any inhibitory ligand, and independent of any chemical or mutagenic alteration.

Voltage-gating and cytosolic Ca2+ activation mechanisms of Arabidopsis two-pore channel AtTPC1

2021 ◽  
Vol 118 (49) ◽  
pp. e2113946118
Author(s):  
Fan Ye ◽  
Lingyi Xu ◽  
Xiaoxiao Li ◽  
Weizhong Zeng ◽  
Ninghai Gan ◽  
...  
Keyword(s):  
Voltage Gating ◽  
Pore Channel ◽  
Voltage Gated ◽  
Gating Mechanism ◽  
Ef Hand ◽  
Activation Mechanisms ◽  
Voltage Gated Ion Channels ◽  
Structural Mechanisms ◽  
Voltage Sensing Domain ◽  
Insight Into

Arabidopsis thaliana two-pore channel AtTPC1 is a voltage-gated, Ca2+-modulated, nonselective cation channel that is localized in the vacuolar membrane and responsible for generating slow vacuolar (SV) current. Under depolarizing membrane potential, cytosolic Ca2+ activates AtTPC1 by binding at the EF-hand domain, whereas luminal Ca2+ inhibits the channel by stabilizing the voltage-sensing domain II (VSDII) in the resting state. Here, we present 2.8 to 3.3 Å cryoelectron microscopy (cryo-EM) structures of AtTPC1 in two conformations, one in closed conformation with unbound EF-hand domain and resting VSDII and the other in a partially open conformation with Ca2+-bound EF-hand domain and activated VSDII. Structural comparison between the two different conformations allows us to elucidate the structural mechanisms of voltage gating, cytosolic Ca2+ activation, and their coupling in AtTPC1. This study also provides structural insight into the general voltage-gating mechanism among voltage-gated ion channels.

scholarly journals Hydrophobic gasket mutation produces gating pore currents in closed human voltage-gated proton channels

2019 ◽  
Vol 116 (38) ◽  
pp. 18951-18961 ◽  
Author(s):  
Richard Banh ◽  
Vladimir V. Cherny ◽  
Deri Morgan ◽  
Boris Musset ◽  
Sarah Thomas ◽  
...  
Keyword(s):  
Proton Channel ◽  
Voltage Gated ◽  
Closed State ◽  
Channel Opening ◽  
Proton Current ◽  
Voltage Dependent ◽  
Current Flows ◽  
Voltage Gated Ion Channels ◽  
Dynamics Simulations ◽  
Voltage Sensing Domain

The hydrophobic gasket (HG), a ring of hydrophobic amino acids in the voltage-sensing domain of most voltage-gated ion channels, forms a constriction between internal and external aqueous vestibules. Cationic Arg or Lys side chains lining the S4 helix move through this “gating pore” when the channel opens. S4 movement may occur during gating of the human voltage-gated proton channel, hHV1, but proton current flows through the same pore in open channels. Here, we replaced putative HG residues with less hydrophobic residues or acidic Asp. Substitution of individuals, pairs, or all 3 HG positions did not impair proton selectivity. Evidently, the HG does not act as a secondary selectivity filter. However, 2 unexpected functions of the HG in HV1 were discovered. Mutating HG residues independently accelerated channel opening and compromised the closed state. Mutants exhibited open–closed gating, but strikingly, at negative voltages where “normal” gating produces a nonconducting closed state, the channel leaked protons. Closed-channel proton current was smaller than open-channel current and was inhibited by 10 μM Zn2+. Extreme hyperpolarization produced a deeper closed state through a weakly voltage-dependent transition. We functionally identify the HG as Val109, Phe150, Val177, and Val178, which play a critical and exclusive role in preventing H+ influx through closed channels. Molecular dynamics simulations revealed enhanced mobility of Arg208 in mutants exhibiting H+ leak. Mutation of HG residues produces gating pore currents reminiscent of several channelopathies.

scholarly journals Large Neurohypophysial Varicosities Amplify Action Potentials: Results from Numerical Simulations

Endocrinology ◽  
2009 ◽  
Vol 150 (6) ◽  
pp. 2829-2836 ◽  
Author(s):  
C. Brad Bennett ◽  
Martin Muschol
Keyword(s):  
Ion Channels ◽  
Numerical Simulations ◽  
Action Potentials ◽  
Calcium Influx ◽  
Physiological Role ◽  
Nonlinear Dependence ◽  
Voltage Gated ◽  
Herring Bodies ◽  
Voltage Gated Ion Channels ◽  
Gated Ion Channels

Axons in the neurohypophysis are known for their “beads on a string” morphology, with numerous in-line secretory swellings lined up along the axon cable. A significant fraction of these secretory swellings, called Herring bodies, is large enough to serve as an identifying feature of the neural lobe in histological sections. Little is known about the physiological role such large axonal swellings might play in neuroendocrine physiology. Using numerical simulations, we have investigated whether large in-line varicosities affect the waveform and propagation of action potentials (APs) along neurohypophysial axons. Due to the strong nonlinear dependence of calcium influx on AP waveforms, such modulation would inevitably affect neuroendocrine release. The parameters for our numerical simulations were matched to established properties of voltage-gated ion channels in neurohypophysial swellings. We find that even a single in-line varicosity can severely depress AP waveforms far upstream in the axonal cable. In contrast, AP depolarization within varicosities becomes amplified. Amplification within varicosities varies in a nontrivial manner with varicosity dimensions, and is most pronounced for diameters close to those of Herring bodies. Overall, we find that large axonal varicosities significantly modulate AP waveforms and their propagation, and do so over large distances. Varicosity size is the main determinant for the observed AP amplification, with the kinetics of voltage-gated ion channels playing a noticeable but secondary role. Our results imply that large varicosities are sites of enhanced hormone release, suggesting that small and large varicosities target different neurohypophysial structures.

scholarly journals Gating mechanism of human N-type voltage-gated calcium channel

2021 ◽  
Author(s):  
Yanli Dong ◽  
Yiwei Gao ◽  
Shuai Xu ◽  
Yuhang Wang ◽  
Zhuoya Yu ◽  
...  
Keyword(s):  
Resting State ◽  
Complex Structure ◽  
The Other ◽  
Nociceptive Transmission ◽  
Voltage Gated ◽  
Gating Mechanisms ◽  
Cryo Electron Microscopy ◽  
Gating Mechanism ◽  
Voltage Sensing Domain ◽  
Structural Insights

N-type voltage-gated calcium (CaV) channels mediate Ca2+ influx at the presynaptic terminals in response to action potential and play vital roles in synaptogenesis, neurotransmitter releasing, and nociceptive transmission. Here we elucidate a cryo-electron microscopy (cryo-EM) structure of the human CaV2.2 complex at resolution of 2.8 Å. This complex structure reveals how the CaV2.2, β1, and α2δ1 subunits are assembled. In our structure, the second voltage-sensing domain (VSD) is stabilized at a resting-state conformation, which is distinct from the other three VSDs of CaV2.2 as well as activated VSDs observed in previous structures of CaV channels. The structure also shows that the intracellular gate formed by S6 helices is closed, and a W-helix from the DII-III linker is determined to act as a blocking-ball that causes closed-state inactivation in CaV2.2. Collectively, our structure provides previously unseen structural insights into fundamental gating mechanisms of CaV channels.

scholarly journals Dendritic Voltage-Gated Ion Channels Regulate the Action Potential Firing Mode of Hippocampal CA1 Pyramidal Neurons

1999 ◽  
Vol 82 (4) ◽  
pp. 1895-1901 ◽  
Author(s):  
Jeffrey C. Magee ◽  
Michael Carruth
Keyword(s):  
Ion Channels ◽  
Action Potential ◽  
Large Amplitude ◽  
Action Potentials ◽  
Pyramidal Neurons ◽  
Burst Firing ◽  
Voltage Gated ◽  
Ca1 Pyramidal Neurons ◽  
Firing Mode ◽  
Voltage Gated Ion Channels

The role of dendritic voltage-gated ion channels in the generation of action potential bursting was investigated using whole cell patch-clamp recordings from the soma and dendrites of CA1 pyramidal neurons located in hippocampal slices of adult rats. Under control conditions somatic current injections evoked single action potentials that were associated with an afterhyperpolarization (AHP). After localized application of 4-aminopyridine (4-AP) to the distal apical dendritic arborization, the same current injections resulted in the generation of an afterdepolarization (ADP) and multiple action potentials. This burst firing was not observed after localized application of 4-AP to the soma/proximal dendrites. The dendritic 4-AP application allowed large-amplitude Na+-dependent action potentials, which were prolonged in duration, to backpropagate into the distal apical dendrites. No change in action potential backpropagation was seen with proximal 4-AP application. Both the ADP and action potential bursting could be inhibited by the bath application of nonspecific concentrations of divalent Ca2+ channel blockers (NiCl and CdCl). Ca2+ channel blockade also reduced the dendritic action potential duration without significantly affecting spike amplitude. Low concentrations of TTX (10–50 nM) also reduced the ability of the CA1 neurons to fire in the busting mode. This effect was found to be the result of an inhibition of backpropagating dendritic action potentials and could be overcome through the coordinated injection of transient, large-amplitude depolarizing current into the dendrite. Dendritic current injections were able to restore the burst firing mode (represented as a large ADP) even in the presence of high concentrations of TTX (300–500 μM). These data suggest the role of dendritic Na+ channels in bursting is to allow somatic/axonal action potentials to backpropagate into the dendrites where they then activate dendritic Ca2+ channels. Although it appears that most Ca2+ channel subtypes are important in burst generation, blockade of T- and R-type Ca2+ channels by NiCl (75 μM) inhibited action potential bursting to a greater extent than L-channel (10 μM nimodipine) or N-, P/Q-type (1 μM ω-conotoxin MVIIC) Ca2+ channel blockade. This suggest that the Ni-sensitive voltage-gated Ca2+ channels have the most important role in action potential burst generation. In summary, these data suggest that the activation of dendritic voltage-gated Ca2+ channels, by large-amplitude backpropagating spikes, provides a prolonged inward current that is capable of generating an ADP and burst of multiple action potentials in the soma of CA1 pyramidal neurons. Dendritic voltage-gated ion channels profoundly regulate the processing and storage of incoming information in CA1 pyramidal neurons by modulating the action potential firing mode from single spiking to burst firing.

scholarly journals Studies of Conorfamide-Sr3 on Human Voltage-Gated Kv1 Potassium Channel Subtypes

Marine Drugs ◽  
2020 ◽  
Vol 18 (8) ◽  
pp. 425
Author(s):  
Estuardo López-Vera ◽  
Luis Martínez-Hernández ◽  
Manuel B. Aguilar ◽  
Elisa Carrillo ◽  
Joanna Gajewiak
Keyword(s):  
Action Potentials ◽  
Inhibitory Concentration ◽  
Molecular Probe ◽  
K Channel ◽  
Dependent Effect ◽  
K Channels ◽  
Voltage Gated ◽  
Pancreatic Cells ◽  
Genes Encoding ◽  
Voltage Gated Ion Channels

Recently, Conorfamide-Sr3 (CNF-Sr3) was isolated from the venom of Conus spurius and was demonstrated to have an inhibitory concentration-dependent effect on the Shaker K+ channel. The voltage-gated potassium channels play critical functions on cellular signaling, from the regeneration of action potentials in neurons to the regulation of insulin secretion in pancreatic cells, among others. In mammals, there are at least 40 genes encoding voltage-gated K+ channels and the process of expression of some of them may include alternative splicing. Given the enormous variety of these channels and the proven use of conotoxins as tools to distinguish different ligand- and voltage-gated ion channels, in this work, we explored the possible effect of CNF-Sr3 on four human voltage-gated K+ channel subtypes homologous to the Shaker channel. CNF-Sr3 showed a 10 times higher affinity for the Kv1.6 subtype with respect to Kv1.3 (IC50 = 2.7 and 24 μM, respectively) and no significant effect on Kv1.4 and Kv1.5 at 10 µM. Thus, CNF-Sr3 might become a novel molecular probe to study diverse aspects of human Kv1.3 and Kv1.6 channels.

Biologically inspired reversible osmotic actuation through voltage-gated ion channels

2012 ◽  
Vol 23 (12) ◽  
pp. 1395-1403 ◽  
Author(s):  
Eric Freeman ◽  
Lisa Weiland
Keyword(s):  
Ion Channels ◽  
Action Potentials ◽  
High Energy ◽  
High Energy Density ◽  
Proof Of Concept ◽  
Biologically Inspired ◽  
Voltage Gated ◽  
Motor Cells ◽  
Voltage Gated Ion Channels ◽  
Gated Ion Channels

This study focuses on the development of a novel high-energy density actuator based on biomimicry principles. The system proposed here draws inspiration from plant motor cells and provides proof of concept for a highly configurable reversible osmotic actuator through the application of voltage-gated ion channels and action potentials. Computational methods are employed to measure the effectiveness of the proposed system in comparison to similar novel actuators.

scholarly journals Voltage-Gated Proton Channels Find Their Dream Job Managing the Respiratory Burst in Phagocytes

Physiology ◽  
2010 ◽  
Vol 25 (1) ◽  
pp. 27-40 ◽  
Author(s):  
Thomas E. DeCoursey
Keyword(s):  
Reactive Oxygen Species ◽  
Nadph Oxidase ◽  
Respiratory Burst ◽  
Proton Channel ◽  
Oxygen Species ◽  
Proton Extrusion ◽  
Proton Channels ◽  
Voltage Gated ◽  
Voltage Gated Ion Channels ◽  
Voltage Sensing Domain

The voltage-gated proton channel bears surprising resemblance to the voltage-sensing domain (S1–S4) of other voltage-gated ion channels but is a dimer with two conduction pathways. The proton channel seems designed for efficient proton extrusion from cells. In phagocytes, it facilitates the production of reactive oxygen species by NADPH oxidase.

scholarly journals The isolated voltage sensing domain of the Shaker potassium channel forms a voltage-gated cation channel

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Juan Zhao ◽  
Rikard Blunck
Keyword(s):  
Transmembrane Helix ◽  
Voltage Sensing ◽  
Shaker Potassium Channel ◽  
Voltage Gated ◽  
Kv Channel ◽  
C Terminus ◽  
Ion Conducting ◽  
Voltage Gated Ion Channels ◽  
Voltage Sensing Domain ◽  
Pore Domain

Domains in macromolecular complexes are often considered structurally and functionally conserved while energetically coupled to each other. In the modular voltage-gated ion channels the central ion-conducting pore is surrounded by four voltage sensing domains (VSDs). Here, the energetic coupling is mediated by interactions between the S4-S5 linker, covalently linking the domains, and the proximal C-terminus. In order to characterize the intrinsic gating of the voltage sensing domain in the absence of the pore domain, the Shaker Kv channel was truncated after the fourth transmembrane helix S4 (Shaker-iVSD). Shaker-iVSD showed significantly altered gating kinetics and formed a cation-selective ion channel with a strong preference for protons. Ion conduction in Shaker-iVSD developed despite identical primary sequence, indicating an allosteric influence of the pore domain. Shaker-iVSD also displays pronounced 'relaxation'. Closing of the pore correlates with entry into relaxation suggesting that the two processes are energetically related.

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