Synthetic Ion Channels Research Articles

Transmembrane Ion Channels: From Natural to Artificial Systems.

Natural channel proteins allow the selective permeation of ions, water or other nutritious entities across bilayer membranes, facilitating various essential physiological functions in living systems. Inspired by nature, chemists endeavor to simulate the structural features and transport behaviors of channel proteins through biomimetic strategies. In this review, we start from introducing the inherent traits of channel proteins such as their crystal structures, functions and mechanisms. Subsequently, different kind of synthetic ion channels including their design principles, dynamic regulations and therapeutic applications were carefully reviewed. Finally, the potential challenges and opportunities in this research field were also carefully discussed. It is anticipated that this review could provide some inspiring ideas and future directions towards the construction of novel bionic ion channels with higher-level structures, properties, functions and practical applications.

Transmembrane Ion Channels: From Natural to Artificial Systems

Natural channel proteins allow the selective permeation of ions, water or other nutritious entities across bilayer membranes, facilitating various essential physiological functions in living systems. Inspired by nature, chemists endeavor to simulate the structural features and transport behaviors of channel proteins through biomimetic strategies. In this review, we start from introducing the inherent traits of channel proteins such as their crystal structures, functions and mechanisms. Subsequently, different kind of synthetic ion channels including their design principles, dynamic regulations and therapeutic applications were carefully reviewed. Finally, the potential challenges and opportunities in this research field were also carefully discussed. It is anticipated that this review could provide some inspiring ideas and future directions towards the construction of novel bionic ion channels with higher‐level structures, properties, functions and practical applications.

Recent Advances in Artificial Anion Channels and Their Selectivity.

Nature performs critical physiological functions using a series of structurally and functionally diverse membrane proteins embedded in cell membranes, in which native ion protein channels modify the electrical potential inside and outside the cell membrane through charged ion movements. Consequently, the cell responds to external stimuli, playing an essential role in various life activities, such as nerve excitation conduction, neurotransmitter release, muscle movement, and control of cell differentiation. Supramolecular artificial channels, which mimic native protein channels in structure and function, adopt unimolecular or self-assembled structures, such as crown ethers, cyclodextrins, cucurbiturils, column arenes, cyclic peptide nanotubes, and metal-organic artificial channels, in channel construction strategies. Owing to the various driving forces involved, artificial synthetic ion channels can be divided into artificial cation and anion channels in terms of ion selectivity. Cation selectivity usually originates from ion coordination, whereas anion selectivity is related to hydrogen bonding, ion pairing, and anion-dipole interactions. Several studies have been conducted on artificial cation channels, and several reviews have summarized them in detail; however, the research on anions is still in the initial stages, and related reviews have rarely been reported. Hence, this article primarily focuses on the recent research on anion channels.

Photo‐activation of Tolane‐based Synthetic Ion Channel for Transmembrane Chloride Transport

While natural channels respond to external stimuli to regulate ion concentration across cell membranes, creating a synthetic version remains challenging. Here, we present a photo‐responsive uncaging technique within an artificial ion channel system, which activates the ion transport process from a transport‐inactive o‐nitrobenzyl‐based caged system. From the comparative ion transport screening, 1b emerged as the most active transporter. Interestingly, its bis(o‐nitrobenzyl) derivative, i.e., protransporter 1b' was inefficient in transporting ions. Detailed transport studies indicated that compound 1b is an anion selective transporter with a prominent selectivity towards chloride ions by following the antiport mechanism. Compound 1b' did not form an ion channel, but after the o‐nitrobenzyl groups were photocleaved, it released 1b, forming a transmembrane ion channel. The channel exhibited an average diameter of 6.5 ± 0.2 Å and a permeability ratio of PCl‐⁄PK+ = 7.3 ± 1.5. The geometry‐optimization of protransporter 1b' indicated significant non‐planarity, corroborating its inefficient self‐assembly. In contrast, the crystal structure of 1b demonstrates strong self‐assembly via the formation of an intermolecular H‐bond. Geometry optimization studies revealed the plausible self‐assembled channel model and the interactions between the channel and chloride ion.

Photo-activation of Tolane-based Synthetic Ion Channel for Transmembrane Chloride Transport.

While natural channels respond to external stimuli to regulate ion concentration across cell membranes, creating a synthetic version remains challenging. Here, we present a photo-responsive uncaging technique within an artificial ion channel system, which activates the ion transport process from a transport-inactive o-nitrobenzyl-based caged system. From the comparative ion transport screening, 1b emerged as the most active transporter. Interestingly, its bis(o-nitrobenzyl) derivative, i.e., protransporter 1b' was inefficient in transporting ions. Detailed transport studies indicated that compound 1b is an anion selective transporter with a prominent selectivity towards chloride ions by following the antiport mechanism. Compound 1b' did not form an ion channel, but after the o-nitrobenzyl groups were photocleaved, it released 1b, forming a transmembrane ion channel. The channel exhibited an average diameter of 6.5 ± 0.2 Å and a permeability ratio of PCl-⁄PK+ = 7.3 ± 1.5. The geometry-optimization of protransporter 1b' indicated significant non-planarity, corroborating its inefficient self-assembly. In contrast, the crystal structure of 1b demonstrates strong self-assembly via the formation of an intermolecular H-bond. Geometry optimization studies revealed the plausible self-assembled channel model and the interactions between the channel and chloride ion.

Using Nanoscale Passports To Understand and Unlock Ion Channels as Gatekeepers of the Cell.

Natural ion channels are proteins embedded in the cell membrane that control many aspects of cell and human physiology by acting as gatekeepers, regulating the flow of ions in and out of cells. Advances in nanotechnology have influenced the methods for studying ion channels in vitro, as well as ways to unlock the delivery of therapeutics by modulating them in vivo. This review provides an overview of nanotechnology-enabled approaches for ion channel research with a focus on the synthesis and applications of synthetic ion channels. Further, the uses of nanotechnology for therapeutic applications are critically analyzed. Finally, we provide an outlook on the opportunities and challenges at the intersection of nanotechnology and ion channels. This work highlights the key role of nanoscale interactions in the operation and modulation of ion channels, which may prompt insights into nanotechnology-enabled mechanisms to study and exploit these systems in the near future.

Artificial transmembrane channel constructed from shape-persistent covalent organic molecular cages capable of ion and small molecule transport

Shape-persistent arylene ethynylene molecular cages have been investigated as transmembrane channels for ions and small molecules. The molecular cages were obtained starting from tetrayne monomers through alkyne metathesis cyclooligomerization. We found these porphyrin-based rigid molecular cages can insert into the lipid bilayer and efficiently transport ions and small molecules (e.g., calcein). Our study reveals longer hydrophobic alkyl chains on the cage molecule promote the channeling efficiency, while shorter and/or more polar side chains impair such activity. Kinetic analysis shows linear correlation between the rate of proton transport and the concentration of the cage, suggesting the active species is likely a monomeric cage. We found that C70-encapsulated cages are nearly inactive for transmembrane ion transportation, indicating that ions are likely transported through the internal cavity of the cage. Discrete shape-persistent organic cages represent highly stable synthetic ion channels or pores, which could have interesting applications in biomimetic signaling and drug delivery.

Redox-Regulated Synthetic Channels: Enabling Reversible Ion Transport by Modulating the Ion-Permeation Pathway.

Natural redox-regulated channel proteins often utilize disulfide bonds as redox sensors for adaptive regulation of channel conformations in response to diverse physiological environments. In this study, we developed novel synthetic ion channels capable of reversibly switching their ion-transport capabilities by incorporating multiple disulfide bonds into artificial systems. X-ray structural analysis and electrophysiological experiments demonstrated that these disulfide-bridged molecules possess well-defined tubular cavities and can be efficiently inserted into lipid bilayers to form artificial ion channels. More importantly, the disulfide bonds in these molecules serve as redox-tunable switches to regulate the formation and disruption of ion-permeation pathways, thereby achieving a transition in the transmembrane transport process between the ON and OFF states.

Redox‐Regulated Synthetic Channels: Enabling Reversible Ion Transport by Modulating the Ion‐Permeation Pathway

AbstractNatural redox‐regulated channel proteins often utilize disulfide bonds as redox sensors for adaptive regulation of channel conformations in response to diverse physiological environments. In this study, we developed novel synthetic ion channels capable of reversibly switching their ion‐transport capabilities by incorporating multiple disulfide bonds into artificial systems. X‐ray structural analysis and electrophysiological experiments demonstrated that these disulfide‐bridged molecules possess well‐defined tubular cavities and can be efficiently inserted into lipid bilayers to form artificial ion channels. More importantly, the disulfide bonds in these molecules serve as redox‐tunable switches to regulate the formation and disruption of ion‐permeation pathways, thereby achieving a transition in the transmembrane transport process between the ON and OFF states.

Confirming Silent Translocation through Nanopores with Simultaneous Single-Molecule Fluorescence and Single-Channel Electrical Recordings.

Most of what is known concerning the luminal passage of materials through nanopores arises from electrical measurements. Whether nanopores are biological, solid-state, synthetic, hybrid, glass-capillary-based, or protein ion channels in cells and tissues, characteristic signatures embedded in the flow of ionic current are foundational to understanding functional behavior. In contrast, this work describes passage through a nanopore that occurs without producing an electrical signature. We refer to the phenomenon as "silent translocation." By definition, silent translocations are invisible to the standard tools of electrophysiology and fundamentally require a simultaneous ancillary measurement technique for positive identification. As a result, this phenomenon has been largely unexplored in the literature. Here, we report on a derivative of Cyanine 5 (sCy5a) that passes through the α-hemolysin (αHL) nanopore silently. Simultaneously acquired single-molecule fluorescence and single-channel electrical recordings from bilayers formed over a closed microcavity demonstrate that translocation does indeed take place, albeit infrequently. We report observations of silent translocation as a function of time, dye concentration, and nanopore population in the bilayer. Lastly, measurement of the translocation rate as a function of applied potential permits estimation of an effective energy barrier for transport through the pore as well as the effective charge on the dye, all in the absence of an information-containing electrical signature.

Mapping metabolic perturbations induced by glutathione activatable synthetic ion channels in human breast cancer cells

Ion channels and transporters play key roles in various biological processes, including cell proliferation and programmed cell death. Recently, we reported that 2,4-dinitrobenzene-sulfonyl-protected N1,N3-dihexy-2-hydroxyisophthalamide (1) forms ion channels upon activation by glutathione (GSH) and results in the induction of apoptosis by depleting the intracellular GSH reservoir in cancer cells. However, the detailed molecular events leading to the induction of apoptosis by these synthetic transport systems in cancer cells still need to be uncovered. Along these lines, we investigated the alterations in cellular metabolites and the associated metabolic pathways by performing untargeted global metabolic profiling of breast cancer cells – MCF-7 – using 1H NMR-based metabolomics. The evaluation of spectral profiles from MCF-7 cells exposed to 1 and their comparison with those corresponding to untreated (control) cells identified 14 significantly perturbed signature metabolites. These metabolites belonged mostly to antioxidant defence, energy metabolism, amino acid biosynthesis, and lipid metabolism pathways and included GSH, o-phosphocholine, malate, and aspartate, to name a few. These results would help us gain deeper insights into the molecular mechanism underlying 1-mediated cytotoxicity of MCF-7 cells and eventually help identify potential novel therapeutic targets for more effective cancer management.

Covalently Linked Hexakis(m-Phenylene Ethynylene) Macrocycles as Molecular Nanotubes.

The construction of nanotubular structures with non-deformable inner pores is of both fundamental and practical significance. Herein we report a strategy for creating molecular nanotubes with defined lengths. Macrocyclic (MC) units based on shape-persistent hexakis(m-phenylene ethynylene) (m-PE) macrocycle MC-1, which are known to stack into hydrogen-bonded tubular assemblies, are tethered by oligo(β-alanine) linkers to give tubular stacks MC-2 and MC-4 that have two and four MC units, respectively. The covalently linked MC units in MC-2 and MC-4 undergo face-to-face stacking through intramolecular non-covalent interactions that further results in the helical stacks of these compounds. Oligomer MC-4 can form potassium and proton channels across lipid bilayers, with the channels being open continuously for over 60 seconds, which is among the longest open durations for synthetic ion channels and indicates that the thermodynamic stability of the self-assembling channels can be drastically enhanced by reducing the number of molecular components involved. This study demonstrates that covalently tethering shape-persistent macrocyclic units is a feasible and reliable approach for building molecular nanotubes that otherwise are difficult to create de novo. The extraordinarily long lifetimes of the ion channels formed by MC-2 and MC-4 suggest the likelihood of constructing the next-generation synthetic ion channels with unprecedented stability.

Covalently Linked Hexakis(m‐Phenylene Ethynylene) Macrocycles as Molecular Nanotubes

AbstractThe construction of nanotubular structures with non‐deformable inner pores is of both fundamental and practical significance. Herein we report a strategy for creating molecular nanotubes with defined lengths. Macrocyclic (MC) units based on shape‐persistent hexakis(m‐phenylene ethynylene) (m‐PE) macrocycle MC‐1, which are known to stack into hydrogen‐bonded tubular assemblies, are tethered by oligo(β‐alanine) linkers to give tubular stacks MC‐2 and MC‐4 that have two and four MC units, respectively. The covalently linked MC units in MC‐2 and MC‐4 undergo face‐to‐face stacking through intramolecular non‐covalent interactions that further results in the helical stacks of these compounds. Oligomer MC‐4 can form potassium and proton channels across lipid bilayers, with the channels being open continuously for over 60 seconds, which is among the longest open durations for synthetic ion channels and indicates that the thermodynamic stability of the self‐assembling channels can be drastically enhanced by reducing the number of molecular components involved. This study demonstrates that covalently tethering shape‐persistent macrocyclic units is a feasible and reliable approach for building molecular nanotubes that otherwise are difficult to create de novo. The extraordinarily long lifetimes of the ion channels formed by MC‐2 and MC‐4 suggest the likelihood of constructing the next‐generation synthetic ion channels with unprecedented stability.

Crown Ether-Based Ion Transporters in Bilayer Membranes.

Bilayer membranes that enhance the stability of the cell are essential for cell survival, separating and protecting the interior of the cell from its external environment. Membrane-based channel proteins are crucial for sustaining cellular activities. However, dysfunction of these proteins would induce serial channelopathies, which could be substituted by artificial ion channel analogs. Crown ethers (CEs) are widely studied in the area of artificial ion channels owing to their intrinsic host-guest interaction with different kinds of organic and inorganic ions. Other advantages such as lower price, chemical stability, and easier modification also make CE a research hotspot in the field of synthetic transmembrane nanopores. And numerous CEs-based membrane-active synthetic ion channels were designed and fabricated in the past decades. Herein, the recent progress of CEs-based synthetic ion transporters has been comprehensively summarized in this review, including their design principles, functional mechanisms, controllable properties, and biomedical applications. Furthermore, this review has been concluded by discussing the future opportunities and challenges facing this research field. It is anticipated that this review could offer some inspiration for the future fabrication of novel CEs-derived ion transporters with more advanced structures, properties, and practical applications.

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.

Nontoxic Artificial Chloride Channel Formation in Epithelial Cells by Isophthalic Acid-Based Small Molecules.

Artificial channels capable of facilitating the transport of Cl- ions across cell membranes while being nontoxic to the cells are rare. Such synthetic ion channels can mimic the functions of membrane transport proteins and, therefore, have the potential to treat channelopathies by replacing defective ion channels. Here we report isophthalic acid-based structurally simple molecules 1 a and 2 a, which self-assemble to render supramolecular nanochannels that allow selective transport of Cl- ions. As evident from the single-crystal X-ray diffraction analysis, the self-assembly is governed by intermolecular hydrogen bonding and π-π stacking interactions. The MD simulation studies for both 1 a and 2 a confirmed the formation of stable Cl- channel assembly in the lipid membrane and Cl- transport through them. The MQAE assay showed the efficacy of the compounds in delivering Cl- ions into cells, and the MTT assays proved that the compounds are nontoxic to cells even at a concentration of 100 μM.

Engineering Polymeric Nanofluidic Membranes for Efficient Ionic Transport: Biomimetic Design, Material Construction, and Advanced Functionalities.

Design elements extracted from biological ion channels guide the engineering of artificial nanofluidic membranes for efficient ionic transport and spawn biomimetic devices with great potential in many cutting-edge areas. In this context, polymeric nanofluidic membranes can be especially attractive because of their inherent flexibility and benign processability, which facilitate massive fabrication and facile device integration for large-scale applications. Herein, the state-of-the-art achievements of polymeric nanofluidic membranes are systematically summarized. Theoretical fundamentals underlying both biological and synthetic ion channels are introduced. The advances of engineering polymeric nanofluidic membranes are then detailed from aspects of structural design, material construction, and chemical functionalization, emphasizing their broad chemical and reticular/topological variety as well as considerable property tunability. After that, this Review expands on examples of evolving these polymeric membranes into macroscopic devices and their potentials in addressing compelling issues in energy conversion and storage systems where efficient ion transport is highly desirable. Finally, a brief outlook on possible future developments in this field is provided.

Structure and Computational Electrophysiology of Ac-LS3, a Synthetic Ion Channel.

Computer simulations are reported on Ac-LS3, a synthetic ion channel, containing 21 residues with a Leu-Ser-Ser-Leu-Leu-Ser-Leu heptad repeat, which forms ions channels upon application of voltage. A hexameric, coiled-coil bundle initially positioned perpendicular to the membrane settled into a stable, tilted structure after 1.5 μs, most likely to improve contacts between the non-polar exterior of the channel and the hydrophobic core of the membrane. Once tilted, the bundle remained in this state during subsequent simulations of nearly 10 μs at voltages ranging from 200 to -100 mV. In contrast, attempts to identify a stable pentameric structure failed, thus supporting the hypothesis that the channel is a hexamer. Results at 100 mV were used to reconstruct the free energy profiles for K+ and Cl- in the channel. This was done by way of several methods in which results of molecular dynamics (MD) simulations were combined with the electrodiffusion model. Two of them developed recently do not require knowledge of the diffusivity. Instead, they utilize one-sided density profiles and committor probabilities. The consistency between different methods is very good, supporting the utility of the newly developed methods for reconstructing free energies of ions in channels. The flux of K+, which accounts for most of the current through the channel, calculated directly from MD matches well the total measured current. However, the current of Cl- is somewhat overestimated, possibly due to a slightly unbalanced force field involving chloride. The current-voltage dependence was also reconstructed by way of a recently developed, efficient method that requires simulations only at a single voltage, yielding good agreement with the experiment. Taken together, the results demonstrate that computational electrophysiology has become a reliable tool for studying how channels mediate ion transport through membranes.

Ion Channels and Transporters as Therapeutic Agents: From Biomolecules to Supramolecular Medicinal Chemistry.

Ion channels and transporters typically consist of biomolecules that play key roles in a large variety of physiological and pathological processes. Traditional therapies include many ion-channel blockers, and some activators, although the exact biochemical pathways and mechanisms that regulate ion homeostasis are yet to be fully elucidated. An emerging area of research with great innovative potential in biomedicine pertains the design and development of synthetic ion channels and transporters, which may provide unexplored therapeutic opportunities. However, most studies in this challenging and multidisciplinary area are still at a fundamental level. In this review, we discuss the progress that has been made over the last five years on ion channels and transporters, touching upon biomolecules and synthetic supramolecules that are relevant to biological use. We conclude with the identification of therapeutic opportunities for future exploration.

Polymerizable channel-like stacks derived from cyclic tetrameric diacetylenes

A small series of cyclic diacetylenes has been synthesized and tested by scanning electron microscopy (SEM), single crystal X-ray diffraction, and atomic force microscopy (AFM) to examine their ability to form arrangements of discrete molecules in the solid state. Examining solid state structures clearly demonstrated propensity of diacetylene tetramers to form channel-like stacks. Remarkably, cyclotetradiyne DA-4 monomer tends to produce two types of nano-tubular motifs (i.e., the channel-like arrangement with solvent matrix inside and unprecedented flattened nano-tube incorporated no solvent guest molecules) which is indicative of the weak C–H/π interactions dominating and directing the stacking in such simplistic diacetylene cyclic systems. Noteworthy, DA-4 molecules may be crystallized from organic solvents into birefringent branchy patterns revealed by Polarized Optical Microscopy (POM) and twisted ribbons of both handedness as evidenced by SEM data. We hypothesize that all these secondary structures may arise from the bundling the individual stacks discovered by single crystal X-ray analysis. Overall, engineering the elongated motifs may open up new opportunities for the future applications such as a rational design of synthetic ion channels and carriers, synthesis of the covalent polymeric nanotubes, etc.