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24 pages, 3596 KiB

Open AccessArticle

Intrinsic Lipid Curvature and Bilayer Elasticity as Regulators of Channel Function: A Comparative Single-Molecule Study

by Mohammad Ashrafuzzaman, Roger E. Koeppe II and Olaf S. Andersen

Int. J. Mol. Sci. 2024, 25(5), 2758; https://doi.org/10.3390/ijms25052758 - 27 Feb 2024

Cited by 2 | Viewed by 855

Abstract

Perturbations in bilayer material properties (thickness, lipid intrinsic curvature and elastic moduli) modulate the free energy difference between different membrane protein conformations, thereby leading to changes in the conformational preferences of bilayer-spanning proteins. To further explore the relative importance of curvature and elasticity [...] Read more.

Perturbations in bilayer material properties (thickness, lipid intrinsic curvature and elastic moduli) modulate the free energy difference between different membrane protein conformations, thereby leading to changes in the conformational preferences of bilayer-spanning proteins. To further explore the relative importance of curvature and elasticity in determining the changes in bilayer properties that underlie the modulation of channel function, we investigated how the micelle-forming amphiphiles Triton X-100, reduced Triton X-100 and the H_II lipid phase promoter capsaicin modulate the function of alamethicin and gramicidin channels. Whether the amphiphile-induced changes in intrinsic curvature were negative or positive, amphiphile addition increased gramicidin channel appearance rates and lifetimes and stabilized the higher conductance states in alamethicin channels. When the intrinsic curvature was modulated by altering phospholipid head group interactions, however, maneuvers that promote a negative-going curvature stabilized the higher conductance states in alamethicin channels but destabilized gramicidin channels. Using gramicidin channels of different lengths to probe for changes in bilayer elasticity, we found that amphiphile adsorption increases bilayer elasticity, whereas altering head group interactions does not. We draw the following conclusions: first, confirming previous studies, both alamethicin and gramicidin channels are modulated by changes in lipid bilayer material properties, the changes occurring in parallel yet differing dependent on the property that is being changed; second, isolated, negative-going changes in curvature stabilize the higher current levels in alamethicin channels and destabilize gramicidin channels; third, increases in bilayer elasticity stabilize the higher current levels in alamethicin channels and stabilize gramicidin channels; and fourth, the energetic consequences of changes in elasticity tend to dominate over changes in curvature. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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18 pages, 2909 KiB

Open AccessArticle

Beta-Barrel Channel Response to High Electric Fields: Functional Gating or Reversible Denaturation?

by Ekaterina M. Nestorovich and Sergey M. Bezrukov

Int. J. Mol. Sci. 2023, 24(23), 16655; https://doi.org/10.3390/ijms242316655 - 23 Nov 2023

Cited by 1 | Viewed by 1025

Abstract

Ion channels exhibit gating behavior, fluctuating between open and closed states, with the transmembrane voltage serving as one of the essential regulators of this process. Voltage gating is a fundamental functional aspect underlying the regulation of ion-selective, mostly α-helical, channels primarily found in [...] Read more.

Ion channels exhibit gating behavior, fluctuating between open and closed states, with the transmembrane voltage serving as one of the essential regulators of this process. Voltage gating is a fundamental functional aspect underlying the regulation of ion-selective, mostly α-helical, channels primarily found in excitable cell membranes. In contrast, there exists another group of larger, and less selective, β-barrel channels of a different origin, which are not directly associated with cell excitability. Remarkably, these channels can also undergo closing, or “gating”, induced by sufficiently strong electric fields. Once the field is removed, the channels reopen, preserving a memory of the gating process. In this study, we explored the hypothesis that the voltage-induced closure of the β-barrel channels can be seen as a form of reversible protein denaturation by the high electric fields applied in model membranes experiments—typically exceeding twenty million volts per meter—rather than a manifestation of functional gating. Here, we focused on the bacterial outer membrane channel OmpF reconstituted into planar lipid bilayers and analyzed various characteristics of the closing-opening process that support this idea. Specifically, we considered the nearly symmetric response to voltages of both polarities, the presence of multiple closed states, the stabilization of the open conformation in channel clusters, the long-term gating memory, and the Hofmeister effects in closing kinetics. Furthermore, we contemplate the evolutionary aspect of the phenomenon, proposing that the field-induced denaturation of membrane proteins might have served as a starting point for their development into amazing molecular machines such as voltage-gated channels of nerve and muscle cells. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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Figure 1
Voltage-dependent closing-opening transitions of OmpF were observed at the single-channel (A) and multichannel (B,C) levels. (A) Typical recordings of ion current through a single trimeric OmpF channel reconstituted into PLMs taken at ±75 mV and ±150 mV applied voltage. Notably, higher voltages lead to the channel closing in approximately three equal but not identical steps. Dashed lines indicate zero current (top) and voltage (bottom) levels. Short-dashed lines designate fully open (L3), one monomer closed (L2), two monomers closed (L1), and residual current (L0) states, with two asterisks (*) highlighting current variations in the L1 state. (B) Raw ion current data from a multichannel system containing around 75 OmpF channels (bottom), subjected to a three-fold repetition of a −200 mV to +200 mV voltage ramp at 1 mHz frequency (top). (C) I-V curves, replotted from the experiments presented in panel (B), at three frequencies (5 mHz, 2 mHz, and 1 mHz), averaged over three ramp cycles, elucidating voltage gating at both polarities. The ion channel recordings were additionally filtered using a 1000 Hz (A) and 100 Hz low-pass Bessel filter (B,C), and data reduction (reduction factor 100, Clampfit, Molecular Devices, San Jose, CA, USA) was applied. Here, the membrane bathing solutions contained 1 M KCl and 5 mM HEPES at pH 7.4. Full article ">Figure 2
The conductance of a multichannel membrane, responding to a voltage ramp from 0.25 mHz to 10 mHz, reveals frequency-dependent sensitivity in its hysteresis curves. (A) Raw data showing ion current across approximately 75 OmpF channels (bottom) in response to a series of repeated 0 to 150 mV voltage ramps at a frequency of 1 mHz (top). (B) Conductance hysteresis curves at different ramp frequencies. Arrows indicate voltage change directions during channel closing and opening within the hysteresis loop. The data represents an average across 3–6 ramp periods. (C) Areas encircled by hysteresis curves from panel (B) as a function of ramp frequency. The data averaged over 3–5 independent experiments with 60–120 OmpF channels. (D) Conductance hysteresis curves from panel (B) after transformation according to Equation (1) in the text. Normalized hysteresis curves show that both the closing and opening branches of the hysteresis curves nearly overlap. (E,F) Gating parameters, the voltage of half-effect <math display="inline"><semantics> <mrow> <msub> <mrow> <mi>V</mi> </mrow> <mrow> <mn>0</mn> </mrow> </msub> </mrow> </semantics></math> (E), and the effective “gating charge” <math display="inline"><semantics> <mrow> <mi>n</mi> </mrow> </semantics></math> (F) obtained by fitting the opening and closing branches of the panel (D) curves to Equation (2) in the text, showing hardly any dependence on the ramp frequency. Full article ">Figure 3
Multichannel conductance relaxation experiment. The experiment was started at the applied voltage of 2.5 mV, then, for durations of 50 s, it was switched to 120 mV, 180 mV, back to 120 mV, and then returned to 2.5 mV as shown in the voltage protocol at the bottom part. Solid lines depict the best-fit single exponentials in the 50 s intervals (the fast component of the opening branch at 100 s is ignored), with the equations displayed in the figure. The lines do not converge to the same value, illustrating the long-term memory of the OmpF voltage-induced closing and opening. The data represent averages from 10 runs. Recordings were filtered using a 100 Hz low-pass Bessel filter, and data reduction (reduction factor 100, Clampfit, Molecular Devices) was applied. Full article ">Figure 4
The current distribution of voltage-induced closing events reveals the variety of conformational states within the closed states of OmpF. (A) An experimental procedure for quantifying the current of the open OmpF ion channel (B), voltage-induced closing event currents (C–E), and residual conductance current (F). To mitigate current variations between channels, data were collected from the same single OmpF channel by repeatedly applying a 150 mV voltage and recording its 3-step closings, similar to those depicted in panel (A), until sufficient statistical data were accumulated. The current histogram from a single open OmpF channel (L3) shown in panel (B), acquired after closings under 150 mV and the subsequent reopenings at 0 mV, displays a narrow current distribution of 651 ± 9 pA. The current histograms in panels C-E of the 1st (L3 → L2), 2nd (L2 → L1), and 3rd (L1 → L0) monomer closings, with the arrows showing transitions, as well as of the residual current (L0) in panel (F), exhibit significantly broader ranges compared to the narrow distribution of the open state current. To ensure easy comparison, panels (B–F) are graphed with the equivalent 250-pA range on the X-axis. Interestingly, the closing event current histograms in panels (C–E) extend beyond one-third of the total channel current, suggesting that the consequences of a single monomer closing could impact nearby monomer conformations and their associated currents. Note that in panel (F), the single Gaussian fit is drawn to guide the eye only as it significantly extends into the unrealistic negative current values. Full article ">Figure 5
Hofmeister effect in OmpF voltage-induced closing. (A) Examples of raw currents through a single OmpF channel at the application of 150 mV. (B) The histograms of the first monomer closing times are fitted by a single exponential function. For illustrative purposes, only the 1 M LiCl (left panel), KCl (middle panel), and RbCl (right panel) data are shown. (C) There is an order of magnitude difference in closing times in the LiCl, NaCl, KCl, RbCl, and CsCl series. (D,E) Multichannel conductance hysteresis curves at 1 mHz ramp frequency (D) show an increase of more than four times in the areas encircled by hysteresis curves (E) in the LiCl, NaCl, KCl, RbCl, CsCl series. (F,G) Multichannel conductance 1 mHz hysteresis curves for different anions. The influence of anions on the OmpF voltage-induced closing and opening (F) is seen as a more than fourfold increase in the areas encircled by hysteresis curves (G) in the KF, KCl, and KBr series. Full article ">Figure 6
Impact of protein cluster insertion on voltage-induced closing. (A,B) A typical multichannel experiment wherein OmpF is exclusively inserted into the PLM as trimeric units, resulting in an average current of 0.23 ± 0.01 nA at 50 mV applied voltage. Notably, upon the application of 150 mV voltage, the channels in this multichannel membrane display a relatively swift and nearly complete closing. OmpF was added into the cis chamber of the bilayer chamber using a 0.22 μg/mL solution following a 1:10,000 stock solution dilution performed approximately eight weeks prior to the experiment. Panel (B) displays a cluster size distribution graph exclusively depicting trimeric OmpF single channel insertions. (C,D) A typical experiment in which about 35% of the total multichannel current was a result of the insertion of OmpF clusters. These clusters comprised thirteen dimers, three trimers, four tetramers, and one octamer, as seen from the cluster distribution graph in panel (D). The remaining current was generated by the insertion of 88 single-channel OmpF trimers. Notably, a significant reduction in the degree of channel closing is observed at 150 mV ((C), right panel). OmpF was added to the cis size of the bilayer chamber using a freshly diluted 0.22 μg/mL solution. (E,F) A typical experiment wherein the insertion of OmpF clusters accounted for over 70% of the total multichannel membrane conductance. These clusters consisted of eight dimers, three trimers, two pentamers, and one decamer (F). Upon applying a voltage of 150 mV ((E), right panel), the effect of voltage on channel closing noticeably diminishes, resulting in a substantial residual conductance that corresponds to approximately 74% of the initial current. OmpF was added to the cis size of the bilayer chamber using a freshly diluted 22 µg/mL solution. The lower panel illustrates the voltage protocol employed for panels (A,C,E). For illustrative purposes, in panel (C), a yellow arrow marks one octamer cluster insertion. In panel (E), orange arrows indicate two pentamer cluster insertions, while a magenta arrow marks one decamer cluster insertion. The same color code is used in panels (B,D,F). The ion channel recordings were filtered using a 50 Hz Bessel filter and data reduction (reduction factor 100, Clampfit, Molecular Devices) was applied. Full article ">Figure 7
One-dimensional cross-section of an energy landscape illustrating voltage-induced channel protein denaturation. Hofmeister series ions are able to change the height of the barrier separating the functional folded state F and the multitude of (partially) unfolded ones U. Full article ">

15 pages, 2071 KiB

Open AccessArticle

Modulation of Voltage-Gating and Hysteresis of Lysenin Channels by Cu²⁺ Ions

by Andrew Bogard, Pangaea W. Finn, Aviana R. Smith, Ilinca M. Flacau, Rose Whiting and Daniel Fologea

Int. J. Mol. Sci. 2023, 24(16), 12996; https://doi.org/10.3390/ijms241612996 - 20 Aug 2023

Cited by 1 | Viewed by 982

Abstract

The intricate voltage regulation presented by lysenin channels reconstituted in artificial lipid membranes leads to a strong hysteresis in conductance, bistability, and memory. Prior investigations on lysenin channels indicate that the hysteresis is modulated by multivalent cations which are also capable of eliciting [...] Read more.

The intricate voltage regulation presented by lysenin channels reconstituted in artificial lipid membranes leads to a strong hysteresis in conductance, bistability, and memory. Prior investigations on lysenin channels indicate that the hysteresis is modulated by multivalent cations which are also capable of eliciting single-step conformational changes and transitions to stable closed or sub-conducting states. However, the influence on voltage regulation of Cu²⁺ ions, capable of completely closing the lysenin channels in a two-step process, was not sufficiently addressed. In this respect, we employed electrophysiology approaches to investigate the response of lysenin channels to variable voltage stimuli in the presence of small concentrations of Cu²⁺ ions. Our experimental results showed that the hysteretic behavior, recorded in response to variable voltage ramps, is accentuated in the presence of Cu²⁺ ions. Using simultaneous AC/DC stimulation, we were able to determine that Cu²⁺ prevents the reopening of channels previously closed by depolarizing potentials and the channels remain in the closed state even in the absence of a transmembrane voltage. In addition, we showed that Cu²⁺ addition reinstates the voltage gating and hysteretic behavior of lysenin channels reconstituted in neutral lipid membranes in which lysenin channels lose their voltage-regulating properties. In the presence of Cu²⁺ ions, lysenin not only regained the voltage gating but also behaved like a long-term molecular memory controlled by electrical potentials. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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13 pages, 2145 KiB

Open AccessArticle

The Complex Proteolipidic Behavior of the SARS-CoV-2 Envelope Protein Channel: Weak Selectivity and Heterogeneous Oligomerization

by Wahyu Surya, Ernesto Tavares-Neto, Andrea Sanchis, María Queralt-Martín, Antonio Alcaraz, Jaume Torres and Vicente M. Aguilella

Int. J. Mol. Sci. 2023, 24(15), 12454; https://doi.org/10.3390/ijms241512454 - 5 Aug 2023

Viewed by 1348

Abstract

The envelope (E) protein is a small polypeptide that can form ion channels in coronaviruses. In SARS coronavirus 2 (SARS-CoV-2), the agent that caused the recent COVID-19 pandemic, and its predecessor SARS-CoV-1, E protein is found in the endoplasmic reticulum–Golgi intermediate compartment (ERGIC), [...] Read more.

The envelope (E) protein is a small polypeptide that can form ion channels in coronaviruses. In SARS coronavirus 2 (SARS-CoV-2), the agent that caused the recent COVID-19 pandemic, and its predecessor SARS-CoV-1, E protein is found in the endoplasmic reticulum–Golgi intermediate compartment (ERGIC), where virion budding takes place. Several reports claim that E protein promotes the formation of “cation-selective channels”. However, whether this term represents specificity to certain ions (e.g., potassium or calcium) or the partial or total exclusion of anions is debatable. Herein, we discuss this claim based on the available data for SARS-CoV-1 and -2 E and on new experiments performed using the untagged full-length E protein from SARS-CoV-2 in planar lipid membranes of different types, including those that closely mimic the ERGIC membrane composition. We provide evidence that the selectivity of the E-induced channels is very mild and depends strongly on lipid environment. Thus, despite past and recent claims, we found no indication that the E protein forms cation-selective channels that prevent anion transport, and even less that E protein forms bona fide specific calcium channels. In fact, the E channel maintains its multi-ionic non-specific neutral character even in concentrated solutions of Ca²⁺ ions. Also, in contrast to previous studies, we found no evidence that SARS-CoV-2 E channel activation requires a particular voltage, high calcium concentrations or low pH, in agreement with available data from SARS-CoV-1 E. In addition, sedimentation velocity experiments suggest that the E channel population is mostly pentameric, but very dynamic and probably heterogeneous, consistent with the broad distribution of conductance values typically found in electrophysiological experiments. The latter has been explained by the presence of proteolipidic channel structures. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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Comparison of SARS E sequences. Sequence comparison between SARS-1 E (accession number P59637) and SARS-2 E (accession number QHZ00401). Transmembrane domain (TMD) is shown in blue. SARS-2 E only differs from SARS-1 E at three mutations and one deletion (underlined in SARS-1 E sequence). The samples used in the present paper (gray rectangle) correspond to SARS-2: full length (E-FL), truncated with a His-tag (His-E-TR) or E-TM. Both E-FL and E-TR contain additional three N-terminal residues (SNA) arising from the N-terminal fusion tag that was cleaved in E-FL, and native Cys residues were mutated to Ala (underlined). For reference, charged residues are indicated as blue (positive) or red (negative) in bold. Full article ">Figure 2
Ohmic character of SARS-2 E-FL in KCl and CaCl2. Example of stable current traces (A,C) and corresponding I–V curves (B,D) for SARS-2 E-FL channels in 500/50 mM KCl at the applied voltages shown in gray below each current trace; (C,D) same as (A,B) but using 500/50 mM CaCl2. The solid line in (B,D) is a linear fit to the experimental data. Full article ">Figure 3
Selectivity (RP) of E protein in different lipid compositions. (A) RP measured in KCl with E-TM 7–38, SARS-1 E-FL or SARS-2 E-FL in neutral DPhPC (black) or negatively charged DPhPS (blue) membranes; (B) RP measured in CaCl2 solutions with SARS-1 E-FL or SARS-2 E-FL in membranes with 20% negatively charged lipids (ERGIC lipid composition). Numbers represent permeability ratios, P+/P− (A) or P−/P+ (B). The horizontal lines indicate the RP of a neutral channel (Vdiff = 0.93 mV, which corresponds to P+/P− = 0.95 (A) or Vdiff = 17.8 mV corresponding to P−/P+ = 3.49 (B)). The concentration gradient was 500/50 mM in all selectivity measurements and all RP values were corrected using the corresponding liquid junction potential from Henderson’s equation to eliminate the contribution of the electrode’s agarose bridges [<a href="#B27-ijms-24-12454" class="html-bibr">27</a>]. Number of experiments was n = 20 (black circle), 15 (black triangle), 16 (black square), 10 (blue circle), 10 (blue triangle), 26 (blue square), 10 (green triangle) and 19 (green square). Error bars indicate ± standard deviation. Data for SARS-1 were reported in [<a href="#B10-ijms-24-12454" class="html-bibr">10</a>,<a href="#B11-ijms-24-12454" class="html-bibr">11</a>]. Full article ">Figure 4
AUC-SV profile of E protein in C14SB detergent micelles. Comparison of c(s) size distribution of (A) E-TM, (B) His-E-TR and (C) E-FL at the indicated detergent-to-protein ratios (DPR, right). The predicted protein molecular weight according to SEDFIT using a common f/f0 for all species is shown with a number (kDa) above each band. Alternatively, the predicted range of S values for various n-oligomeric states (n is shown on the right of the bar) are shown as colored bars. These ranges were calculated as described in the <a href="#sec4-ijms-24-12454" class="html-sec">Section 4</a>; (D–G) two-dimensional plots c(s, f/f0) corresponding to E-TM (D), E-TR (E,F) and E-FL (G) at the DPRs indicated. Full article ">

14 pages, 1835 KiB

Open AccessArticle

Triplin: Mechanistic Basis for Voltage Gating

by Marco Colombini, Patrick Liu and Chase Dee

Int. J. Mol. Sci. 2023, 24(14), 11473; https://doi.org/10.3390/ijms241411473 - 14 Jul 2023

Cited by 1 | Viewed by 879

Abstract

The outer membrane of Gram-negative bacteria contains a variety of pore-forming structures collectively referred to as porins. Some of these are voltage dependent, but weakly so, closing at high voltages. Triplin, a novel bacterial pore-former, is a three-pore structure, highly voltage dependent, with [...] Read more.

The outer membrane of Gram-negative bacteria contains a variety of pore-forming structures collectively referred to as porins. Some of these are voltage dependent, but weakly so, closing at high voltages. Triplin, a novel bacterial pore-former, is a three-pore structure, highly voltage dependent, with a complex gating process. The three pores close sequentially: pore 1 at positive potentials, 2 at negative and 3 at positive. A positive domain containing 14 positive charges (the voltage sensor) translocates through the membrane during the closing process, and the translocation is proposed to take place by the domain entering the pore and thus blocking it, resulting in the closed conformation. This mechanism of pore closure is supported by kinetic measurements that show that in the closing process the voltage sensor travels through most of the transmembrane voltage before reaching the energy barrier. Voltage-dependent blockage of the pores by polyarginine, but not by a 500-fold higher concentrations of polylysine, is consistent with the model of pore closure, with the sensor consisting mainly of arginine residues, and with the presence, in each pore, of a complementary surface that serves as a binding site for the sensor. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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Review

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17 pages, 2708 KiB

Open AccessReview

Counter-Intuitive Features of Particle Dynamics in Nanopores

by Alexander M. Berezhkovskii and Sergey M. Bezrukov

Int. J. Mol. Sci. 2023, 24(21), 15923; https://doi.org/10.3390/ijms242115923 - 3 Nov 2023

Cited by 1 | Viewed by 739

Abstract

Using the framework of a continuous diffusion model based on the Smoluchowski equation, we analyze particle dynamics in the confinement of a transmembrane nanopore. We briefly review existing analytical results to highlight consequences of interactions between the channel nanopore and the translocating particles. [...] Read more.

Using the framework of a continuous diffusion model based on the Smoluchowski equation, we analyze particle dynamics in the confinement of a transmembrane nanopore. We briefly review existing analytical results to highlight consequences of interactions between the channel nanopore and the translocating particles. These interactions are described within a minimalistic approach by lumping together multiple physical forces acting on the particle in the pore into a one-dimensional potential of mean force. Such radical simplification allows us to obtain transparent analytical results, often in a simple algebraic form. While most of our findings are quite intuitive, some of them may seem unexpected and even surprising at first glance. The focus is on five examples: (i) attractive interactions between the particles and the nanopore create a potential well and thus cause the particles to spend more time in the pore but, nevertheless, increase their net flux; (ii) if the potential well-describing particle-pore interaction occupies only a part of the pore length, the mean translocation time is a non-monotonic function of the well length, first increasing and then decreasing with the length; (iii) when a rectangular potential well occupies the entire nanopore, the mean particle residence time in the pore is independent of the particle diffusivity inside the pore and depends only on its diffusivity in the bulk; (iv) although in the presence of a potential bias applied to the nanopore the “downhill” particle flux is higher than the “uphill” one, the mean translocation times and their distributions are identical, i.e., independent of the translocation direction; and (v) fast spontaneous gating affects nanopore selectivity when its characteristic time is comparable to that of the particle transport through the pore. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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12 pages, 1324 KiB

Open AccessReview

Mechanisms of PIEZO Channel Inactivation

by Zijing Zhou and Boris Martinac

Int. J. Mol. Sci. 2023, 24(18), 14113; https://doi.org/10.3390/ijms241814113 - 14 Sep 2023

Cited by 3 | Viewed by 2598

Abstract

PIEZO channels PIEZO1 and PIEZO2 are the newly identified mechanosensitive, non-selective cation channels permeable to Ca²⁺. In higher vertebrates, PIEZO1 is expressed ubiquitously in most tissues and cells while PIEZO2 is expressed more specifically in the peripheral sensory neurons. PIEZO channels [...] Read more.

PIEZO channels PIEZO1 and PIEZO2 are the newly identified mechanosensitive, non-selective cation channels permeable to Ca²⁺. In higher vertebrates, PIEZO1 is expressed ubiquitously in most tissues and cells while PIEZO2 is expressed more specifically in the peripheral sensory neurons. PIEZO channels contribute to a wide range of biological behaviors and developmental processes, therefore driving significant attention in the effort to understand their molecular properties. One prominent property of PIEZO channels is their rapid inactivation, which manifests itself as a decrease in channel open probability in the presence of a sustained mechanical stimulus. The lack of the PIEZO channel inactivation is linked to various mechanopathologies emphasizing the significance of studying this PIEZO channel property and the factors affecting it. In the present review, we discuss the mechanisms underlying the PIEZO channel inactivation, its modulation by the interaction of the channels with lipids and/or proteins, and how the changes in PIEZO inactivation by the channel mutations can cause a variety of diseases in animals and humans. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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Essential domains and residues for PIEZO inactivation. (A) Cap domain and inner helix are highlighted in the 3D structure of mouse PIEZO1 (PDB: 6BPZ). Critical amino acids in mouse PIEZO1, together with their positions, are highlighted in red; positions of amino acids in human PIEZO1 are in grey. The right panel presents a zoom-in diagram of the inner helix with the critical amino acids. (B) Structure overview of wild-type PIEZO1 and PIEZO2, or chimeric PIEZO1 or PIEZO2 fused with the ‘Cap’ domain (C terminal extracellular domain, CED) from each other, is shown in the left panel. Inactivation of PIEZO1 is faster than PIEZO2 (right panel, up). Swapping the CED exchanges the inactivation constant of the channels, as P1-P2CED inactivates faster than the P2-P1CED (right panel, down). The figure is adapted from (Wu et al., 2017) under CC BY-NC-ND 4.0. (C) Manipulating the hydrophobic gate by mutating L2475/V2476 of mouse PIEZO1 significantly reduces inactivation compared to the wild-type. Figure is adopted from [<a href="#B28-ijms-24-14113" class="html-bibr">28</a>] under CC BY-NC-ND 4.0. Full article ">Figure 2
Interacting proteins may explain the slow-inactivating PIEZO1 currents in native cells. (A) Indentation-induced whole-cell currents in N2A cells overexpressing mouse PIEZO1 (Left) or native PIEZO1 current in the mouse embryonic stem cells (Right). Native PIEZO1 shows a slow inactivating kinetics. (B) 3D structure of PIEZO1-MDFIC Complex. The C terminus of MDFIC is shown in green. MDFIC inserts into PIEZO1′s pore module and stays near the inner helix, as detailed in the right panel. (C) HEK cells overexpressing PIEZO1 show a rapid inactivating current (Left). Upon co-expression of MDFIC, the inactivation is largely removed (Right). Figures are adopted from [<a href="#B54-ijms-24-14113" class="html-bibr">54</a>] and [<a href="#B55-ijms-24-14113" class="html-bibr">55</a>] under CC BY-NC-ND 4.0. Full article ">

26 pages, 3737 KiB

Open AccessReview

Gating of β-Barrel Protein Pores, Porins, and Channels: An Old Problem with New Facets

by Lauren A. Mayse and Liviu Movileanu

Int. J. Mol. Sci. 2023, 24(15), 12095; https://doi.org/10.3390/ijms241512095 - 28 Jul 2023

Cited by 5 | Viewed by 1706

Abstract

β barrels are ubiquitous proteins in the outer membranes of mitochondria, chloroplasts, and Gram-negative bacteria. These transmembrane proteins (TMPs) execute a wide variety of tasks. For example, they can serve as transporters, receptors, membrane-bound enzymes, as well as adhesion, structural, and signaling elements. [...] Read more.

β barrels are ubiquitous proteins in the outer membranes of mitochondria, chloroplasts, and Gram-negative bacteria. These transmembrane proteins (TMPs) execute a wide variety of tasks. For example, they can serve as transporters, receptors, membrane-bound enzymes, as well as adhesion, structural, and signaling elements. In addition, multimeric β barrels are common structural scaffolds among many pore-forming toxins. Significant progress has been made in understanding the functional, structural, biochemical, and biophysical features of these robust and versatile proteins. One frequently encountered fundamental trait of all β barrels is their voltage-dependent gating. This process consists of reversible or permanent conformational transitions between a large-conductance, highly permeable open state and a low-conductance, solute-restrictive closed state. Several intrinsic molecular mechanisms and environmental factors modulate this universal property of β barrels. This review article outlines the typical signatures of voltage-dependent gating. Moreover, we discuss recent developments leading to a better qualitative understanding of the closure dynamics of these TMPs. Full article

(This article belongs to the Special Issue Membrane Channels: Mechanistic Insights)

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