scholarly journals New Insights on the Voltage Dependence of the KCa3.1 Channel Block by Internal TBA

2004 ◽  
Vol 124 (4) ◽  
pp. 333-348 ◽  
Author(s):  
Umberto Banderali ◽  
Hélène Klein ◽  
Line Garneau ◽  
Manuel Simoes ◽  
Lucie Parent ◽  
...  
Keyword(s):  
Transmembrane Potential ◽  
Voltage Dependence ◽  
Selectivity Filter ◽  
Channel Structure ◽  
Potential Profile ◽  
Ammonium Ion ◽  
Channel Pore ◽  
Internal Medium ◽  
K Concentration ◽  
External K

We present in this work a structural model of the open IKCa (KCa3.1) channel derived by homology modeling from the MthK channel structure, and used this model to compute the transmembrane potential profile along the channel pore. This analysis showed that the selectivity filter and the region extending from the channel inner cavity to the internal medium should respectively account for 81% and 16% of the transmembrane potential difference. We found however that the voltage dependence of the IKCa block by the quaternary ammonium ion TBA applied internally is compatible with an apparent electrical distance δ of 0.49 ± 0.02 (n = 6) for negative potentials. To reconcile this observation with the electrostatic potential profile predicted for the channel pore, we modeled the IKCa block by TBA assuming that the voltage dependence of the block is governed by both the difference in potential between the channel cavity and the internal medium, and the potential profile along the selectivity filter region through an effect on the filter ion occupancy states. The resulting model predicts that δ should be voltage dependent, being larger at negative than positive potentials. The model also indicates that raising the internal K+ concentration should decrease the value of δ measured at negative potentials independently of the external K+ concentration, whereas raising the external K+ concentration should minimally affect δ for concentrations >50 mM. All these predictions are born out by our current experimental results. Finally, we found that the substitutions V275C and V275A increased the voltage sensitivity of the TBA block, suggesting that TBA could move further into the pore, thus leading to stronger interactions between TBA and the ions in the selectivity filter. Globally, these results support a model whereby the voltage dependence of the TBA block in IKCa is mainly governed by the voltage dependence of the ion occupancy states of the selectivity filter.

scholarly journals Two Stable, Conducting Conformations of the Selectivity Filter in Shaker K+ Channels

2005 ◽  
Vol 125 (6) ◽  
pp. 619-629 ◽  
Author(s):  
Jill Thompson ◽  
Ted Begenisich
Keyword(s):  
Voltage Dependence ◽  
Relative Proportion ◽  
Selectivity Filter ◽  
K Channel ◽  
K Channels ◽  
High Affinity ◽  
K Concentration ◽  
The One ◽  
Low K ◽  
Two Populations

We have examined the voltage dependence of external TEA block of Shaker K+ channels over a range of internal K+ concentrations from 2 to 135 mM. We found that the concentration dependence of external TEA block in low internal K+ solutions could not be described by a single TEA binding affinity. The deviation from a single TEA binding isotherm was increased at more depolarized membrane voltages. The data were well described by a two-component binding scheme representing two, relatively stable populations of conducting channels that differ in their affinity for external TEA. The relative proportion of these two populations was not much affected by membrane voltage but did depend on the internal K+ concentration. Low internal K+ promoted an increase in the fraction of channels with a low TEA affinity. The voltage dependence of the apparent high-affinity TEA binding constant depended on the internal K+ concentration, becoming almost voltage independent in 5 mM. The K+ sensitivity of these low- and high-affinity TEA states suggests that they may represent one- and two-ion occupancy states of the selectivity filter, consistent with recent crystallographic results from the bacterial KcsA K+ channel. We therefore analyzed these data in terms of such a model and found a large (almost 14-fold) difference between the intrinsic TEA affinity of the one-ion and two-ion modes. According to this analysis, the single ion in the one-ion mode (at 0 mV) prefers the inner end of the selectivity filter twofold more than the outer end. This distribution does not change with internal K+. The two ions in the two-ion mode prefer to occupy the inner end of the selectivity filter at low K+, but high internal K+ promotes increased occupancy of the outer sites. Our analysis further suggests that the four K+ sites in the selectivity filter are spaced between 20 and 25% of the membrane electric field.

scholarly journals In silico Exploration of Interactions Between Potential COVID-19 Antiviral Treatments and the Pore of the hERG Potassium Channel—A Drug Antitarget

2021 ◽  
Vol 8 ◽  
Author(s):  
Ehab Al-Moubarak ◽  
Mohsen Sharifi ◽  
Jules C. Hancox
Keyword(s):  
Potassium Channel ◽  
In Silico ◽  
Open Channel ◽  
Antiviral Drugs ◽  
Selectivity Filter ◽  
Channel Structure ◽  
Herg Channel ◽  
Closed Channel ◽  
Aromatic Residues ◽  
Channel Pore

Background: In the absence of SARS-CoV-2 specific antiviral treatments, various repurposed pharmaceutical approaches are under investigation for the treatment of COVID-19. Antiviral drugs considered for this condition include atazanavir, remdesivir, lopinavir-ritonavir, and favipiravir. Whilst the combination of lopinavir and ritonavir has been previously linked to prolongation of the QTc interval on the ECG and risk of torsades de pointes arrhythmia, less is known in this regard about atazanavir, remdesivir, and favipiravir. Unwanted abnormalities of drug-induced QTc prolongation by diverse drugs are commonly mediated by a single cardiac anti-target, the hERG potassium channel. This computational modeling study was undertaken in order to explore the ability of these five drugs to interact with known determinants of drug binding to the hERG channel pore.Methods: Atazanavir, remdesivir, ritonavir, lopinavir and favipiravir were docked to in silico models of the pore domain of hERG, derived from cryo-EM structures of hERG and the closely related EAG channel.Results: Atazanavir was readily accommodated in the open hERG channel pore in proximity to the S6 Y652 and F656 residues, consistent with published experimental data implicating these aromatic residues in atazanavir binding to the channel. Lopinavir, ritonavir, and remdesivir were also accommodated in the open channel, making contacts in a model-dependent fashion with S6 aromatic residues and with residues at the base of the selectivity filter/pore helix. The ability of remdesivir (at 30 μM) to inhibit the channel was confirmed using patch-clamp recording. None of these four drugs could be accommodated in the closed channel structure. Favipiravir, a much smaller molecule, was able to fit within the closed channel and could adopt multiple binding poses in the open channel, but with few simultaneous interactions with key binding residues. Only favipiravir and remdesivir showed the potential to interact with lateral pockets below the selectivity filter of the channel.Conclusions: All the antiviral drugs studied here can, in principle, interact with components of the hERG potassium channel canonical binding site, but are likely to differ in their ability to access lateral binding pockets. Favipiravir's small size and relatively paucity of simultaneous interactions may confer reduced hERG liability compared to the other drugs. Experimental structure-function studies are now warranted to validate these observations.

scholarly journals Selectivity filter modalities and rapid inactivation of the hERG1 channel

2020 ◽  
Vol 117 (6) ◽  
pp. 2795-2804 ◽  
Author(s):  
Williams E. Miranda ◽  
Kevin R. DeMarco ◽  
Jiqing Guo ◽  
Henry J. Duff ◽  
Igor Vorobyov ◽  
...  
Keyword(s):  
Qualitative Agreement ◽  
Molecular Mechanisms ◽  
Selectivity Filter ◽  
Channel Structure ◽  
Wild Type ◽  
Selective Transport ◽  
Markov State Model ◽  
Computational Electrophysiology ◽  
Herg1 Channel ◽  
External K

The human ether-á-go-go–related gene (hERG1) channel conducts small outward K+ currents that are critical for cardiomyocyte membrane repolarization. The gain-of-function mutation N629D at the outer mouth of the selectivity filter (SF) disrupts inactivation and K+-selective transport in hERG1, leading to arrhythmogenic phenotypes associated with long-QT syndrome. Here, we combined computational electrophysiology with Markov state model analysis to investigate how SF-level gating modalities control selective cation transport in wild-type (WT) and mutant (N629D) hERG1 variants. Starting from the recently reported cryogenic electron microscopy (cryo-EM) open-state channel structure, multiple microseconds-long molecular-dynamics (MD) trajectories were generated using different cation configurations at the filter, voltages, electrolyte concentrations, and force-field parameters. Most of the K+ permeation events observed in hERG1-WT simulations occurred at microsecond timescales, influenced by the spontaneous dehydration/rehydration dynamics at the filter. The SF region displayed conductive, constricted, occluded, and dilated states, in qualitative agreement with the well-documented flickering conductance of hERG1. In line with mutagenesis studies, these gating modalities resulted from dynamic interaction networks involving residues from the SF, outer-mouth vestibule, P-helices, and S5–P segments. We found that N629D mutation significantly stabilizes the SF in a state that is permeable to both K+ and Na+, which is reminiscent of the SF in the nonselective bacterial NaK channel. Increasing the external K+ concentration induced “WT-like” SF dynamics in N629D, in qualitative agreement with the recovery of flickering currents in experiments. Overall, our findings provide an understanding of the molecular mechanisms controlling selective transport in K+ channels with a nonconventional SF sequence.

scholarly journals Affinity and Location of an Internal K+ Ion Binding Site in Shaker K Channels

2001 ◽  
Vol 117 (5) ◽  
pp. 373-384 ◽  
Author(s):  
Jill Thompson ◽  
Ted Begenisich
Keyword(s):  
Electric Field ◽  
Binding Sites ◽  
Voltage Dependence ◽  
Ion Binding ◽  
K Channels ◽  
Shaker Channels ◽  
Ion Binding Sites ◽  
K Concentration ◽  
Shaker Potassium Channels ◽  
External K

We have examined the interaction between TEA and K+ ions in the pore of Shaker potassium channels. We found that the ability of external TEA to antagonize block of Shaker channels by internal TEA depended on internal K+ ions. In contrast, this antagonism was independent of external K+ concentrations between 0.2 and 40 mM. The external TEA antagonism of internal TEA block increased linearly with the concentration of internal K+ ions. In addition, block by external TEA was significantly enhanced by increases in the internal K+ concentration. These results suggested that external TEA ions do not directly antagonize internal TEA, but rather promote increased occupancy of an internal K+ site by inhibiting the emptying of that site to the external side of the pore. We found this mechanism to be quantitatively consistent with the results and revealed an intrinsic affinity of the site for K+ ions near 65 mM located ∼7% into the membrane electric field from the internal end of the pore. We also found that the voltage dependence of block by internal TEA was influenced by internal K+ ions. The TEA site (at 0 internal K+) appeared to sense ∼5% of the field from the internal end of the pore (essentially colocalized with the internal K+ site). These results lead to a refined picture of the number and location of ion binding sites at the inner end of the pore in Shaker K channels.

scholarly journals An Anionic Ryanoid, 10-O-succinoylryanodol, Provides Insights into the Mechanisms Governing the Interaction of Ryanoids and the Subsequent Altered Function of Ryanodine-receptor Channels

2003 ◽  
Vol 121 (6) ◽  
pp. 551-561 ◽  
Author(s):  
Bhavna Tanna ◽  
William Welch ◽  
Luc Ruest ◽  
John L. Sutko ◽  
Alan J. Williams
Keyword(s):  
Ryanodine Receptor ◽  
Transmembrane Potential ◽  
Voltage Dependence ◽  
Major Influence ◽  
Ionic Interactions ◽  
Receptor Affinity ◽  
Flexible Molecule ◽  
Ion Translocation ◽  
Conductance States ◽  
Conductance State

We have investigated the interactions of a novel anionic ryanoid, 10-O-succinoylryanodol, with individual mammalian cardiac muscle ryanodine receptor channels under voltage clamp conditions. As is the case for all ryanoids so far examined, the interaction of 10-O-succinoylryanodol with an individual RyR channel produces profound alterations in both channel gating and rates of ion translocation. In the continued presence of the ryanoid the channel fluctuates between periods of normal and modified gating, indicating a reversible interaction of the ligand with its receptor. Unlike the majority of ryanoids, we observe a range of different fractional conductance states of RyR in the presence of 10-O-succinoylryanodol. We demonstrate that 10-O-succinoylryanodol is a very flexible molecule and propose that each fractional conductance state arises from the interaction of a different conformer of the ryanoid molecule with the RyR channel. The probability of channel modification by 10-O-succinoylryanodol is dependent on the transmembrane holding potential. Comparison of the voltage dependence of channel modification by this novel anionic ryanoid with previous data obtained with cationic and neutral ryanoids reveals that the major influence of transmembrane potential on the probability of RyR channel modification by ryanoids results from an alteration in receptor affinity. These investigations also demonstrate that the charge of the ryanoid has a major influence on the rate of association of the ligand with its receptor indicating that ionic interactions are likely to be involved in this reaction.

scholarly journals A model for the effects of potential and external K+ concentration on the Cs+ blocking of inward rectification

1980 ◽  
Vol 30 (1) ◽  
pp. 199-204 ◽  
Author(s):  
S. Ciani ◽  
S. Krasne ◽  
S. Hagiwara
Keyword(s):  
Inward Rectification ◽  
K Concentration ◽  
External K

Delay of desensitization onset by potassium ion in voltage-clamped frog muscle fibers

1985 ◽  
Vol 249 (5) ◽  
pp. C435-C446 ◽  
Author(s):  
A. A. Manthey
Keyword(s):  
Voltage Clamp ◽  
Acetylcholine Receptors ◽  
Transmembrane Potential ◽  
Direct Action ◽  
Direct Interaction ◽  
Frog Muscle ◽  
Average Potential ◽  
Voltage Dependent ◽  
Source Voltage ◽  
K Concentration

Increase in extracellular K+ concentration causes delay in desensitization onset during prolonged application of carbamylcholine to the postjunctional membrane in muscle. This could be due to a direct action of K+ on acetylcholine receptors or to some change in the receptors related to K+-induced effects on transmembrane potential. The question of direct vs. voltage-dependent action of K+ was investigated in frog muscle (Rana pipiens) using a point-source voltage clamp. In conductance measurements first without voltage control, desensitization rate in bath media containing 33 mM K+ was -0.198 s-1 among fibers showing an average potential of -30 mV and -0.104 s-1 in 165 mM K+ where the average potential was -2 mV, a decrease of 47%. By comparison, in voltage-clamp tests at a nominal holding potential of +20 mV, increasing extracellular K+ from 33 to 165 mM caused a decrease of 61% in desensitization rate from -0.151 to -0.059 s-1. Another series in 165 mM K+ at a holding level of +10 mV showed a decrease of 67% to a rate of 0.047 s-1. It is concluded that increases in extracellular K+ can delay desensitization onset independently of effects on transmembrane potential. It is suggested that this could result from a direct interaction of K+ with sites on the outer receptor moiety or within channels, but probably not at the inner membrane face, if the latter are considered in equilibrium with bulk intracellular K+.

scholarly journals Ion Transport in Hydrodictyon africanum

1967 ◽  
Vol 50 (6) ◽  
pp. 1607-1625 ◽  
Author(s):  
J. A. Raven
Keyword(s):  
Free Energy ◽  
Low Temperature ◽  
Electrical Potential ◽  
Bathing Solution ◽  
Electrical Potential Difference ◽  
Energy Gradient ◽  
Na Efflux ◽  
K Concentration ◽  
Net Fluxes ◽  
External K

The concentrations of K, Na, and Cl in the cytoplasm and vacuole, the tracer fluxes of these ions into and out of the cenocyte, and the electrical potential difference between bathing solution and vacuole and cytoplasm, have been measured in Hydrodictyon africanum. If the ions were acted on solely by passive electrochemical forces, a net efflux of K and Cl and a net influx of Na would be expected. Tracer fluxes indicate a net influx of K and Cl and efflux of Na in the light; these net fluxes are consequently active, with an obligate link to metabolism. The effects of darkness and low temperature indicate that most of the tracer K and Cl influx and Na efflux are linked to metabolism, while the corresponding tracer fluxes in the direction of the free energy gradient are not. Ouabain specifically inhibits the metabolically linked portions of tracer K influx and Na efflux. Alterations in the external K concentration have similar effects on metabolically mediated K influx and Na efflux. It would appear that K influx and Na efflux are linked, at least in the light.

Uptake of K+ by frog gastric mucosa from submucosal side and acid secretory rate.

1977 ◽  
Vol 232 (3) ◽  
pp. E294
Author(s):  
N Takeguchi ◽  
I Horikoshi ◽  
M Hattori
Keyword(s):  
Gastric Mucosa ◽  
Linear Relation ◽  
Secretory Rate ◽  
Co2 Gas ◽  
Control Value ◽  
Different Types ◽  
K Concentration ◽  
Mucosal Side ◽  
External K

The K+ content in frog gastric mucosa (K+) was measured as a function of the submucosal K+ concentration ([K+sm]) in the absence of K+ on the mucosal side. The (K+) in HCO3- buffer with 95% O2-5% CO2 gas showed that the removal of external K+ induced a 27% K+ loss from the control value of 5 mM K+sm, that there was no linear relation between (K+) and [K+sm, and that the change in the (K+) versus the [K+sm] was hyperbolic, indicating that there are two different types of K+ in the mucosa: bound and free K+.

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