Permeable Cation Research Articles

A critical role for altered red cell cation permeability in pathogenesis of sickle cell disease and other haemolytic anaemias.

The aetiology of sickle cell disease is well known, but pathogenesis is complicated and details remain uncertain. A thorough understanding may suggest novel ways for designing more effective therapies. One area of importance, covered here in Nader et al., is the altered cation permeability of sickle cells and how the co-ordinated operation of a number of membrane transport proteins contributes to disease progression, all driven by the initial event of HbS polymerisation. There are echoes here of the cation leaks of hereditary stomatocytosis. Nader et al. propose a central role for PIEZO1, a novel mechanosensitive channel found in red cells, which may be aberrantly activated in sickle cells following HbS polymerisation and which may have potential as a novel target for future chemotherapies. Commentary on: Nader et al. Piezo1 activation augments sickling propensity and the adhesive properties of sickle red blood cells in a calcium-dependent manner. Br J Haematol 2023;202:657-668.

Structural basis of sensory receptor evolution in octopus.

Chemotactile receptors (CRs) are a cephalopod-specific innovation that allow octopuses to explore the seafloor via 'taste by touch'1. CRs diverged from nicotinic acetylcholine receptors to mediate contact-dependent chemosensation of insoluble molecules that do not readily diffuse in marine environments. Here we exploit octopus CRs to probe the structural basis of sensory receptor evolution. We present the cryo-electron microscopy structure of an octopus CR and compare it with nicotinic receptors to determine features that enable environmental sensation versus neurotransmission. Evolutionary, structural and biophysical analyses show that thechannel architecture involved in cation permeation and signal transduction is conserved. By contrast, the orthosteric ligand-binding site is subject to diversifying selection, thereby mediating the detection of new molecules. Serendipitous findings in the cryo-electron microscopy structure reveal that the octopus CR ligand-binding pocket is exceptionally hydrophobic, enabling sensation of greasy compounds versus the small polar molecules detected by canonical neurotransmitter receptors. These discoveries provide a structural framework for understanding connections between evolutionary adaptations at the atomic level and the emergence of new organismal behaviour.

Mechanistic Insights on Permeation of Water over Iron Cations in Nanoporous Silicon Oxide Films for Selective H2 and O2 Evolution

Electrocatalysts encapsulated by an ultrathin and semipermeable oxide layer offer a promising avenue for efficient, selective, and cost-effective production of hydrogen through photoelectrochemical water splitting. This architecture is especially attractive for Z-scheme water splitting, for which a nanoporous oxide film can be leveraged to mitigate undesired, yet kinetically facile, reactions involving redox shuttles, such as aqueous iron cations, by limiting transport of these species to catalytically active sites. In this work, molecular dynamics simulations were combined with electrochemical measurements to provide a mechanistic understanding of permeation of water and Fe(III)/Fe(II) redox shuttles through nanoporous SiO2 films. It is shown that even for SiO2 pores with a width as small as 0.8 nm, water does not experience any energy barrier for permeating into the pores due to a favorable interaction with hydrophilic silanol groups on the oxide surface. In contrast, permeation of Fe(III) and Fe(II) into microporous SiO2 pores is limited due to high energy barriers, which stem from a combination of distortion and dehydration of the second and third ion solvation shells. Our simulations and experimental results show that SiO2 coatings can effectively mitigate undesired Fe(III)/Fe(II) redox reactions at underlying electrodes by attenuating permeation of iron cations, while allowing water to permeate and thus participate in water splitting reactions. In a broader context, our study demonstrates that selectivity of solvated cations can be manipulated by controlling the pore size and surface chemistry of oxide films.

A cooperative knock-on mechanism underpins Ca2+-selective cation permeation in TRPV channels.

The selective exchange of ions across cellular membranes is a vital biological process. Ca2+-mediated signaling is implicated in a broad array of physiological processes in cells, while elevated intracellular concentrations of Ca2+ are cytotoxic. Due to the significance of this cation, strict Ca2+ concentration gradients are maintained across the plasma and organelle membranes. Therefore, Ca2+ signaling relies on permeation through selective ion channels that control the flux of Ca2+ ions. A key family of Ca2+-permeable membrane channels is the polymodal signal-detecting transient receptor potential (TRP) ion channels. TRP channels are activated by a wide variety of cues including temperature, small molecules, transmembrane voltage, and mechanical stimuli. While most members of this family permeate a broad range of cations non-selectively, TRPV5 and TRPV6 are unique due to their strong Ca2+ selectivity. Here, we address the question of how some members of the TRPV subfamily show a high degree of Ca2+ selectivity while others conduct a wider spectrum of cations. We present results from all-atom molecular dynamics simulations of ion permeation through two Ca2+-selective and two non-selective TRPV channels. Using a new method to quantify permeation cooperativity based on mutual information, we show that Ca2+-selective TRPV channel permeation occurs by a three-binding site knock-on mechanism, whereas a two-binding site knock-on mechanism is observed in non-selective TRPV channels. Each of the ion binding sites involved displayed greater affinity for Ca2+ over Na+. As such, our results suggest that coupling to an extra binding site in the Ca2+-selective TRPV channels underpins their increased selectivity for Ca2+ over Na+ ions. Furthermore, analysis of all available TRPV channel structures shows that the selectivity filter entrance region is wider for the non-selective TRPV channels, slightly destabilizing ion binding at this site, which is likely to underlie mechanistic decoupling.

Ion permeation pathway within the internal pore of P2X receptor channels.

P2X receptor channels are trimeric ATP-activated ion channels expressed in neuronal and non-neuronal cells that are attractive therapeutic targets for human disorders. Seven subtypes of P2X receptor channels have been identified in mammals that can form both homomeric and heteromeric channels. P2X1-4 and P2X7 receptor channels are cation-selective, whereas P2X5 has been reported to have both cation and anion permeability. P2X receptor channel structures reveal that each subunit is comprised of two transmembrane helices, with both N-and C-termini on the intracellular side of the membrane and a large extracellular domain that contains the ATP binding sites at subunit interfaces. Recent structures of ATP-bound P2X receptors with the activation gate open reveal the unanticipated presence of a cytoplasmic cap over the central ion permeation pathway, leaving lateral fenestrations that may be largely buried within the membrane as potential pathways for ions to permeate the intracellular end of the pore. In the present study, we identify a critical residue within the intracellular lateral fenestrations that is readily accessible to thiol-reactive compounds from both sides of the membrane and where substitutions influence the relative permeability of the channel to cations and anions. Taken together, our results demonstrate that ions can enter or exit the internal pore through lateral fenestrations that play a critical role in determining the ion selectivity of P2X receptor channels.

Lipid peroxidation increases membrane tension, Piezo1 gating, and cation permeability to execute ferroptosis

The ongoing metabolic and microbicidal pathways that support and protect cellular life generate potentially damaging reactive oxygen species (ROS). To counteract damage, cells express peroxidases, which are antioxidant enzymes that catalyze the reduction of oxidized biomolecules. Glutathione peroxidase 4 (GPX4) is the major hydroperoxidase specifically responsible for reducing lipid peroxides; this homeostatic mechanism is essential, and its inhibition causes a unique type of lytic cell death, ferroptosis. The mechanism(s) that lead to cell lysis in ferroptosis, however, are unclear. We report that the lipid peroxides formed during ferroptosis accumulate preferentially at the plasma membrane. Oxidation of surface membrane lipids increased tension on the plasma membrane and led to the activation of Piezo1 and TRP channels. Oxidized membranes thus became permeable to cations, ultimately leading to the gain of cellular Na+ and Ca2+ concomitant with loss of K+. These effects were reduced by deletion of Piezo1 and completely inhibited by blocking cation channel conductance with ruthenium red or 2-aminoethoxydiphenyl borate (2-APB). We also found that the oxidation of lipids depressed the activity of the Na+/K+-ATPase, exacerbating the dissipation of monovalent cation gradients. Preventing the changes in cation content attenuated ferroptosis. Altogether, our study establishes that increased membrane permeability to cations is a critical step in the execution of ferroptosis and identifies Piezo1, TRP channels, and the Na+/K+-ATPase as targets/effectors of this type of cell death.

Variant spectrum of PIEZO1 and KCNN4 in Japanese patients with dehydrated hereditary stomatocytosis

Hereditary stomatocytosis (HSt) is a type of congenital hemolytic anemia caused by abnormally increased cation permeability of erythrocyte membranes. Dehydrated HSt (DHSt) is the most common subtype of HSt and is diagnosed based on clinical and laboratory findings related to erythrocytes. PIEZO1 and KCNN4 have been recognized as causative genes, and many related variants have been reported. We analyzed the genomic background of 23 patients from 20 Japanese families suspected of having DHSt using a target capture sequence and identified pathogenic/likely pathogenic variants of PIEZO1 or KCNN4 in 12 families.

Insight into the role of ionicity in the desalination and separation of a graphene oxide membrane

To provide a comprehensive understanding on the role of ionicity of solutes and polarity of the graphene oxide (GO) surface in permeabilities, a near-practical lab-scale cross-flow-type apparatus was used herein to systematically examine the separation behaviors of representative inorganic cations (alkaline and alkaline earth cations and the counter anions) and dye molecules through GO membrane (GOM). It was found that cation separations through a dried GOM of tens nanometer thickness receives strong influence from electrostatic interactions. As the result, both cationic permeation rate and water flux obey a linear increasing relationship with the strength of cationic power whereas each of the monovalent (Li+, Na+, K+, Rb+, Cs+) and bivalent (Mg2+, Ca2+, Sr2+) cation groups have a different increasing gradient. Accumulation adsorption through ion-exchange in the GO interlayers induces an increase in penetration rejection of organic dyes at the cost of water flux due to membrane blockade. The cation separation mechanism was understood through intercalation structure analysis and detailed using molecular dynamic simulations. The research results disclosed a dominant ion-exchange mechanism and the crucial role of ionicity of solutes in separation. These findings fill up the gap in understanding differences of salts and dyes migration within GOM, being worthwhile for future membrane material design and the references of various water treatment applications using GOM.

Dynamic communication between TRPC3 and IP3R shape Ca2+ signaling signatures.

Physical and functional coupling between TRPC channels and IP3 receptors (IP3R) has been proposed as a pivotal mechanism in cellular Ca2+ signaling. Here we set out to explore the impact of TRPC3-IP3R communication at the level of spatiotemporal organisation of Ca2+ signaling. The low affinity Ca2+ sensor R-GECO1.2 was fused as a reporter to the C-terminus of TRPC3 and the channel function of this construct was modified specifically by point mutations. Cytoplasmic Ca2+ changes at the TRPC3 channels expressed in HEK293 cells originated from both the entry of Ca2+ through the TRPC channel and Ca2+ mobilization via IP3R. Local Ca2+ changes within the TRPC3 signaling domain were predominantly biphasic with IP3R-dependent initial Ca2+ transients, while exclusively monophasic signals were recorded when all three IP3R isoforms were lacking. Abrogation of Ca2+ entry through TRPC3 by mutational impairment of Ca2+ permeability (E630Q), cation permeation (E630K), or DAG sensitivity (G652A), promoted local Ca2+ oscillations. Ca2+ signals at E630Q, E630K, and G652A channels featured initial Ca2+ transients along with oscillatory activity. By contrast, oscillations and initial Ca2+ transients, were virtually lacking, when the TRPC3 channels were sensitized by subthreshold stimulation of phospholipase C. TIRF imaging provided evidence for dynamic colocalization of TRPC3 and IP3R. Our results suggest that TRPC3-mediated Ca2+ entry controls IP3R activity at ER-PM junctions to shape Ca2+ signaling signatures and enable specificity of downstream signaling.

Identification of specific sphingolipid-bama interactions using coarse-grained molecular dynamics simulation of the C. crescentus outer membrane.

Sphingolipids (SLs) have been primarily studied in eukaryotic cell membranes, where they modulate fundamental cellular processes ranging from apoptosis to cytoskeletal reorganization. It has recently been discovered that many bacteria also synthesize sphingolipids, but their role in bacterial membranes is poorly understood. One example of a gram-negative bacteria with sphingolipids in its outer membrane is C. crescentus. Previous studies showed that the absence of SLs in the outer membrane increases resistance to cationic polymyxin antibiotics, permeability, and susceptibility to antimicrobials like bacitracin while inducing an unfolded-protein stress response. The machinery responsible for the folding of many outer membrane proteins (OMPs) is the β-barrel assembly machinery (BAM complex). In the present study, we used coarse-grained molecular dynamics (CG-MD) simulations to understand the possible interactions between SLs and the BAM complex to determine if SLs regulate BamA activity by direct interaction. The C. crescentus BamA protein was homology modeled in two conformations: open- and closed-gate. Martini models of the rough lipopolysaccharide (RLPS), smooth lipopolysaccharide (SLPS), and anionic SLs of C. crescentus were built. We carried out CG-MD simulations of BamA embedded in a model of a C. crescentus bacterial membrane in the closed and open conformations. Afterward, the density distribution of SLs with respect to BamA was analyzed, using the CG-MD analysis toolkit Nougat from Brannigan Lab.

Investigation of nonselective cation permeation through CNGA1 channels by MD simulations.

Mammalian cyclic nucleotide-gated (CNG) channels are crucial components of numerous signal transduction pathways, most classically in the visual and olfactory sensory systems. The transmembrane domain of CNG acts as nonselective cation channels, which are opened by the binding of cGMP in the ligand binding domain. Recently, high-resolution cryo-EM structures of human CNGA1 and heteromeric CNGA1/CNGB1 channels became available, but the detailed mechanism governing non-selective cation permeation in CNG remains elusive.

Analysis of cation permeation mechanism of ChR2 by molecular dynamics simulation.

The biomembranes separate intra- and extra-cellular. Permeation of small molecules via the biomembrane is mainly regulated by transport proteins. The ion channel transports ions along an electrochemical potential gradient. Channelrhodopsin-2 (ChR2) is a light-activated cation channel and the first widely adopted optogenetic tool. The structure of ChR2 is a homo-dimer of 7-transmembrane G-protein with covalently bound retinal. ChR2 absorbs blue light with maximum absorption and action spectrum at 480 nm. When the all-trans-retinal complex absorbs a photon, it changes into 13-cis-retinal and induces a conformation change of the protein to permeate cations. It is also known that a mutant ChR2 changes ion permeability. However, the ion permeation mechanism of ChR2 has not been clarified yet. In this study, we have performed molecular dynamics simulations of ChR2 with different conditions and compared them to examine the mechanism of ion permeability.

Cu(II)-Triggered Ion Channel Properties of a 2,2'-Bipyridine-Modified Amphotericin B.

The development of stimuli-responsive synthetic channels that open and close in response to physical and chemical changes in the surrounding environment has attracted attention because of their potential bioapplications such as sensing, drug release, antibiotics, and molecular manipulation tools to control membrane transport in cells. Metal coordination is ideal as a stimulus for stimuli-responsive channels because it allows for reversible gating behavior through the addition and removal of metal ions and fine-tuning of channel structure through coordination geometry defined by the type of the metal ion and ligand. We have previously reported on transition metal-ion dependent ion permeability control of Amphotericin B (AmB) modified with a metal coordination site, 2,2'-bipyridine ligand (bpy-AmB). AmB is one of the polyene macrolide antibiotics, and it is known that the interaction between AmB and ergosterol molecules is required for AmB channel formation. In contrast, the Cu2+ coordination to the bpy moiety of bpy-AmB induces formation of Ca2+ ion-permeable channels in the ergosterol-free POPC membrane. However, the details of bpy-AmB properties such as channel stability, ion selectivity, pore size, and the effect of ergosterol on channel formation remain unclear. Here, we investigate bpy-AmB channels triggered by transition metal coordination in POPC or ergosterol-containing POPC liposomes using an HPTS assay, electrophysiological measurements, and time-resolved UV-vis spectral measurements. These analyses reveal that bpy-AmB channels triggered by Cu2+ ions are more stable and have larger pore sizes than the original AmB channels and enable efficient permeation of various cations. We believe that our channel design will lead to the construction of metal coordination-triggered synthetic ion channels.

HC067047 as a potent TRPV4 inhibitor repairs endotoxemia colonic injury

Colonic injury causes severe inflammation during systemic infections in patients with endotoxemia. The prevention of colonic injury could effectively reduce the progression of endotoxemia. We investigated the protective effects and detailed mechanisms of the TRPV4 inhibitor HC067047 in the treatment of colonic injury caused by endotoxemia. An LPS-induced endotoxemia colonic injury model was used to assess the in vivo effects of HC067047. Colon slices were detected by hematoxylin and eosin (HE) staining and immunofluorescence assays. Spectrophotometry was used to determine the levels of MDA, calcium, GSH, and GSSG. Alterations in oxidative stress/mitophagy/inflammatory pyroptosis-related markers were evaluated by Q-PCR and western blot assays. HC067047 reduced the body weight loss and spleen weight index of endotoxemic mice and partly recovered the normal morphology of the colonic mucous layer. As an inhibitor of the calcium permeant cation channel, HC067047 suppressed the phosphorylation of the CAMKIIɑ protein and levels of MDA and calcium, upregulated the ratio of GSH/GSSG, shortened the expression of oxidative stress-related proteins, and enhanced the expression of the anti-oxidative protein CAT in damaged colon tissues. Additionally, HC067047 maintained normal mitochondrial functions in endotoxemia colons by promoting mitochondrial fusion and biosynthesis and suppressing mitochondrial fission and the PINK/Parkin/mitophagy pathway. HC067047 potently blocked inflammatory pyroptosis and protected the colonic tight junction barrier. HC067047 restores endotoxemia colons against oxidative stress, mitophagy, inflammatory pyroptosis, and colonic barrier dysfunction. Hence, HC067047 therapy may be potentially useful in the treatment of colonic injury in endotoxemia.

The SARS-CoV-2 accessory protein Orf3a is not an ion channel, but does interact with trafficking proteins.

The severe acute respiratory syndrome associated coronavirus 2 (SARS-CoV-2) and SARS-CoV-1 accessory protein Orf3a colocalizes with markers of the plasma membrane, endocytic pathway, and Golgi apparatus. Some reports have led to annotation of both Orf3a proteins as viroporins. Here, we show that neither SARS-CoV-2 nor SARS-CoV-1 Orf3a form functional ion conducting pores and that the conductances measured are common contaminants in overexpression and with high levels of protein in reconstitution studies. Cryo-EM structures of both SARS-CoV-2 and SARS-CoV-1 Orf3a display a narrow constriction and the presence of a positively charged aqueous vestibule, which would not favor cation permeation. We observe enrichment of the late endosomal marker Rab7 upon SARS-CoV-2 Orf3a overexpression, and co-immunoprecipitation with VPS39. Interestingly, SARS-CoV-1 Orf3a does not cause the same cellular phenotype as SARS-CoV-2 Orf3a and does not interact with VPS39. To explain this difference, we find that a divergent, unstructured loop of SARS-CoV-2 Orf3a facilitates its binding with VPS39, a HOPS complex tethering protein involved in late endosome and autophagosome fusion with lysosomes. We suggest that the added loop enhances SARS-CoV-2 Orf3a's ability to co-opt host cellular trafficking mechanisms for viral exit or host immune evasion.

Synthetic K+ Channels Constructed by Rebuilding the Core Modules of Natural K+ Channels in an Artificial System.

Different types of natural K+ channels share similar core modules and cation permeability characteristics. In this study, we have developed novel artificial K+ channels by rebuilding the core modules of natural K+ channels in artificial systems. All the channels displayed high selectivity for K+ over Na+ and exhibited a selectivity sequence of K+ ≈Rb+ during the transport process, which is highly consistent with the cation permeability characteristics of natural K+ channels. More importantly, these artificial channels could be efficiently inserted into cell membranes and mediate the transmembrane transport of K+ , disrupting the cellular K+ homeostasis and eventually triggering the apoptosis of cells. These findings demonstrate that, by rebuilding the core modules of natural K+ channels in artificial systems, the structures, transport behaviors, and physiological functions of natural K+ channels can be mimicked in synthetic channels.

Synthetic K+Channels Constructed by Rebuilding the Core Modules of Natural K+Channels in an Artificial System

AbstractDifferent types of natural K+channels share similar core modules and cation permeability characteristics. In this study, we have developed novel artificial K+channels by rebuilding the core modules of natural K+channels in artificial systems. All the channels displayed high selectivity for K+over Na+and exhibited a selectivity sequence of K+≈Rb+during the transport process, which is highly consistent with the cation permeability characteristics of natural K+channels. More importantly, these artificial channels could be efficiently inserted into cell membranes and mediate the transmembrane transport of K+, disrupting the cellular K+homeostasis and eventually triggering the apoptosis of cells. These findings demonstrate that, by rebuilding the core modules of natural K+channels in artificial systems, the structures, transport behaviors, and physiological functions of natural K+channels can be mimicked in synthetic channels.

Clinical Practice Guidelines for the Use of Transcranial Direct Current Stimulation in Psychiatry.

Clinical Practice Guidelines for the Use of Transcranial Direct Current Stimulation in Psychiatry.

Ion dehydration using magnetic fields and impacts on permeability across RO membranes

Magnetic fields have been used as a mitigation process for controlling mineral scaling in diverse applications, including membrane systems. Changes in scale formation have been attributed, in part, to changes in ion hydration. The objective of this study was to evaluate how passage through a multi-directional magnetic field affected the permeability of relevant cations and anions through a seawater reverse osmosis membrane (SW30HR). Changes in ion permeability were evaluated as a function of flow velocity through the magnetic system and described in terms of changes in the ion hydration and in turn changes in the energy penalty that a given ion must incur to permeate across the RO membrane. Ion permeability across the SW30HR membrane increased after having passed through the magnetic system. Increases in ion permeability became most pronounced once a flow velocity through the magnetic system was ≥35 cm/s and were a function of the ion charge density for ions of the same valence. Increases in ion permeability were attributed to dehydration of the ions by the magnetic system resulting in a reduction in the energy penalty required for the relevant ions to transport across the membrane barrier.

Janus membrane with tailored upper and lower surface charges for ion penetration manipulation in high-performance nanofiltration

Conventional homogeneously charged nanofiltration (NF) membranes showed limited capabilities in ion transmembrane behavior control, which hindered the ion separation performances. In this study, novel Janus membranes were developed to manipulate the penetration of monovalent and divalent ions in NF. Graphene oxide nanosheets with enhanced positive charge (PGO) and negative charge (NGO) were successfully synthesized and assembled in designed orders and proportions to construct the Janus membranes with desired upper and lower surface charges. The positively charged upper surface and the negatively charged lower surface of the PN Janus membrane showed high retention of divalent cations but high permeation of monovalent cations; when the mass ratio of PGO to NGO was 1–5, the PN-1/5 membrane indicated a significant Li+/Mg2+ separation factor at 23.5. On the other hand, the NP-4/2 Janus membrane composed of negatively charged upper layer and positively charged lower layer indicated the most promising indiscriminate divalent ion removal efficiency (>80%) as compared with the literature. The molecular dynamics simulation theoretically proved the ion penetration behavior manipulation by the Janus membranes were feasible. This study proposed a facile strategy to dramatically improve the performances of NF in Mg2+/Li+ separation and indiscriminate divalent ion removal.