Interaction of polypeptide neurotoxins with a receptor site associated with voltage-sensitive sodium channels

Anthopleurin A, a polypeptide toxin from the Pacific sea anemone Anthopleura xanthogrammica, enhances persistent activation of voltage-sensitive sodium channels by the alkaloid toxins veratridine and batrachotoxin with K0.5 = 20 nM. This effect is inhibited by depolarization. There is a close correlation between enhancement of sodium channel activation and block of [125I]scorpion toxin binding by unlabeled scorpion toxin, sea anemone toxin II from Anemonia sulcata, and Anthopleurin A, indicating that these three polypeptide toxins interact with a common receptor site in modifying sodium channel function. Photo-activable derivatives of scorpion toxin label a single Mr approximately 250,000 polypeptide chain at the polypeptide toxin receptor site. Labeling is blocked by unlabeled scorpion toxin or depolarization and is not observed in variant neuroblastoma clones, which lack sodium channels. These results identify a protein component of the polypeptide toxin receptor site of voltage-sensitive sodium channels.     

Halogen Bonding Heteroditopic Materials for Cooperative Sodium Iodide Binding and Extraction

A series of novel heteroditopic halogen bonding (XB) receptor functionalised silica based materials, containing mono- and bis-iodotriazole benzo-15-crown-5 groups are investigated for the cooperative binding and extraction of sodium halide ion-pair species from aqueous solution. Characterisation of the XB materials by CHN elemental analysis, ¹³C CP/MAS NMR and ATR-FTIR spectroscopies confirms and quantifies the successful incorporation of the ion-pair receptor frameworks to the silica material. ICP-MS solid-liquid extraction studies demonstrate the bidentate XB functionalised material is capable of NaI extraction from water. Importantly, cooperative XB-mediated sodium halide ion-pair binding is determined to be crucial to the material's extraction capabilities, impressively demonstrating a two-fold enhancement in sodium iodide extraction efficiency relative to a heteroditopic hydrogen bonding receptor functionalised silica material analogue.     

Chalcogen Bond Mediated Enhancement of Cooperative Ion-Pair Recognition

A series of heteroditopic receptors containing halogen bond (XB) and unprecedented chalcogen bond (ChB) donors integrated into a 3,5-bis-triazole pyridine structure covalently linked to benzo-15-crown-5 ether motifs exhibit remarkable cooperative recognition of halide anions. Multi-nuclear ¹H, ¹³C, ¹²⁵Te and ¹⁹F NMR, ion pair binding investigations reveal sodium cation–benzo-crown ether binding dramatically enhances the recognition of bromide and iodide halide anions, with the chalcogen bonding heteroditopic receptor notably displaying the largest enhancement of halide binding strength of over two hundred-fold, in comparison to the halogen bonding and hydrogen bonding heteroditopic receptor analogues. DFT calculations suggest crown ether sodium cation complexation induces a polarisation of the sigma hole of ChB and XB heteroditopic receptor donors as a significant contribution to the origin of the unique cooperativity exhibited by these systems.     

An electron-rich three dimensional receptor based on a calixarene-tetrathiafulvalene assembly

The synthesis of a calix[4]arene scaffold persubstituted with four redox-active tetrathiafulvalene (TTF) moieties at the lower rim is described. This assembly strongly binds sodium cation, and the binding process is accompanied by a conformational change of the receptor, as shown from NMR titration and by an X-ray diffraction led on the complex. This dynamic behavior remarkably results in a modification of the electrochemical response of TTF probes, which behave independently after sodium complexation.     

Acetylcholine Sensor Based on Ion Sensitive Field Effect Transistor and Acetylcholine Receptor

Abstract

A new type of acetylcholine sensor was made with an Ion Sensitive Field Effect Transistor (ISFET) and acetylcholine receptor. The acetylcholine receptor was fixed on a polyvinylbutyral membrane which covered the ISFET gate. When acetylcholine was injected into this system, the differential gate output voltage gradually Shifted to the positive side and reached a constant value. This response was due to the positive charge of acetylcholine. A linear relationship was obtained between the initial rate of the differential gate output voltage change and the logarithmic value of the acetylcholine concentration. Acetylcholine was fixed in the range 0.1-10μM. When the acetylcholine receptor was immobilized with the lipid membrane, the response was amplified with both the positive charge of acetylcholine and sodium ion flux through the acetylcholine receptor's channel. Therefore, the difference in the differential voltage between the acetylcholine receptor-ISFET systems with and without the lipid membrane was caused by sodium ion flux through the acetylcholine receptor's channel.     

Chalcogen Bond Mediated Enhancement of Cooperative Ion‐Pair Recognition

A series of heteroditopic receptors containing halogen bond (XB) and unprecedented chalcogen bond (ChB) donors integrated into a 3,5‐bis‐triazole pyridine structure covalently linked to benzo‐15‐crown‐5 ether motifs exhibit remarkable cooperative recognition of halide anions. Multi‐nuclear ¹H, ¹³C, ¹²⁵Te and ¹⁹F NMR, ion pair binding investigations reveal sodium cation–benzo‐crown ether binding dramatically enhances the recognition of bromide and iodide halide anions, with the chalcogen bonding heteroditopic receptor notably displaying the largest enhancement of halide binding strength of over two hundred‐fold, in comparison to the halogen bonding and hydrogen bonding heteroditopic receptor analogues. DFT calculations suggest crown ether sodium cation complexation induces a polarisation of the sigma hole of ChB and XB heteroditopic receptor donors as a significant contribution to the origin of the unique cooperativity exhibited by these systems.     

Sodium and pH responsive hydrogel formation by the supramolecular system calix[4]pyrrole derivative/tetramethylammonium cation

We report and rationalize the formation of gels, responsive to sodium cations or pH changes, in aqueous media by a supramolecular system constituted of an aryl extended calix[4]pyrrole receptor and the tetramethylammonium guest.     

A halogen-bonding-based heteroditopic receptor for alkali metal halides

A new heteroditopic receptor for alkali metal halides has been designed and synthesized. It is comprised of a well-established motif for cation binding and a motif for halogen-bonding-based anion recognition processes. The single-crystal X-ray structure of the complex between the heteroditopic receptor and sodium iodide is reported. Thanks to the cooperativity of metal coordination and the strong I-...I halogen bonding, the ion pair is fully separated. The boosting effect of the binding of the anion through halogen bonding on the coordination of the cation by the receptor has been proved also in solution by NMR experiments. The selectivity of the new heterotopic receptor toward different alkali metal halides has been tested by ESI mass experiments.     

Dependence of turbidity oscillations of self-oscillating polymers on the selective recognition of the crown ether receptors contained in the polymer chain

The turbidity oscillations of self-oscillating polymers in the Belousov-Zhabotinsky (BZ) reaction system depending on the crown ether receptors contained in the polymer network have been studied. The three monomers are copolymerized, namely, N-isopropylacrylamide, the metal catalyst monomer for the BZ reaction, and the crown ether receptor monomer, to prepare the self-oscillating polymers used in this study. The turbidity oscillations are characterized by monitoring the transmittance of the polymer solution in the BZ reaction system at a specific wavelength of 570 nm. The oscillations are varied by crown ether receptors used in the polymerization process, i.e., BCAm(6) or BCAm(5), for the selective recognition of specific cations between potassium and sodium ions in the solution. The selective recognition of the BCAm receptors in the polymer chain for the two ions has brought out a variation in the turbidity oscillations by a change in the hydrophilicity of the polymer chain. The oscillations of the polymer solution composed of the BCAm(5) receptor are more influenced by sodium ion, while the polymer solution of BCAm(6) receptor is affected by potassium ion. However, the oscillation patterns of the redox changes obtained by these solution systems look much alike despite the differences in the polymer chain by crown ether receptors and cations of bromate used for the BZ reaction.     

Facilitated transport of sodium or potassium chloride across vesicle membranes using a ditopic salt-binding macrobicycle

A synthetic receptor, with an ability to bind sodium or potassium chloride as a contact ion-pair, is shown to effectively transport either salt across vesicle membranes. Significant transport is observed even when the transporter: phospholipid ratio is as low as 1:2500. Chloride efflux from unilamellar vesicles is monitored using a chloride selective electrode. Mechanistic studies indicate that the facilitated efflux is due to the uncomplexed transporter diffusing into the vesicle and the transporter-salt complex diffusing out. Vesicle influx experiments are also reported, where the facilitated influx of chloride and sodium ions into vesicles is observed directly by 35Cl and 23Na NMR, respectively.     

Synthesis,structutre and biological activity of substituted [16α,17α]cyclopropapregn-4-ene-3,20-diones

Aryldiazomethanes generated by vacuum thermolysis of arenecarbaldehyde tosylhydrazone sodium salts react stereospecifically with 16-dehydropregnenolone acetate. Subsequent decomposition of the pyrazoline adducts yielded hitherto unknown substituted [16α,17α]-cyclopropaprogesterones bearing the aryl substituents at the position 3′. Interaction of such cyclopropaprogesterones with progesterone receptor from rat uterine cytosol was studied.     

A combined extraction and DFT study on the complexation of H3O+ with a hexaarylbenzene-based receptor

Abstract  

By means of theoretical calculations at density functional level, the complex structure of a hexaarylbenzene-based receptor with Na⁺ was derived. In the resulting complex having C ₃ symmetry, the sodium cation synergistically interacts with the hydrophilic polar ethereal oxygen fence and with the central hydrophobic benzene bottom of the parent receptor via cation–π interaction.     

Lithium cation enhances anion binding in a tripodal phosphine oxide-based ditopic receptor

A tripodal ditopic receptor presents H-bond donors and a phosphine oxide to potential guests. In the idealized binding conformation, an endohedral P=O functionality provides enhanced halide binding in the presence of lithium with the greatest ΔΔG° observed for bromide, while minimal changes in K(a) are observed in the presence of sodium.     

New Bis- and Tris- Benzo-Crown Ether Receptor Molecules Containing Respectively Ter-Anisyl and Tren Bridging Linkages

Abstract

New bis- and tris- benzo-crown ether receptor molecules containing respectively a rigid ter-anisyl bridge and a Schiff base tren linkage have been prepared. Preliminary coordination studies of these ligands with potassium, sodium and copper guest cations have led to the formation of monometallic and hetero-polymetallic complexes.     

A carbohydrate-conjugated deep cavitand permits observation of caviplexes in human serum

A deep cavitand was covalently modified with carbohydrates to provide solubility in biologically relevant environments and to investigate its receptor function. Specifically, a tetrakis(β-D-glucosyl) cavitand (1) that was soluble in neutral water or acid/base-buffered solutions was synthesized, and it formed complexes with hydrophobic small molecules. Extraction of the cavitand into aqueous sodium dodecyl sulfate micelles as simple membrane mimetics increased the scope of guests bound by 1 beyond that observed in only aqueous media. Complex formation was also detected in human serum. The findings show the functional compatibility of the receptor in both micelle-bound and serum-soluble forms.     

Improved detection of the histamine receptor on human peripheral blood mononuclear cells by crosslinking using tritiated as compared with radioiodinated histamine

In order to detect histamine receptors on the surface of human peripheral blood mononuclear cells, the cells were incubated in the presence of radiolabelled histamine and then the bifunctional crosslinker disuccimidyl suberate was added in various concentrations. They were then solubilized with sodium dodecyl sulphate, boiled, reduced and the lysate separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Both 3H and 125I-radiolabelled ligands bound to a 16 kDa band, to be defined although a much clearer and obviously unequivocal signal was obtained with 3H-labelled histamine. This molecule migrated with the same mass on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as a 16 kDa subunit which had been purified on a histamine affinity column from Triton X-100 solubilized mononuclear cells, indicating it to be the ligand-binding subunit for the histamine receptor on these cells. For 3H, fluorography with Entensify was required to obtain an autoradiographic signal. Although 3H took much longer to give a signal than 125I, the considerable background, artefacts and heavy lane trailing seen with [125I] histamine were completely abrogated when [3H]histamine was used. In addition, the distinction between specific and nonspecific binding was more clearly seen using [3H]histamine. The modifications reported here which improve signal detection for 3H should encourage the use of tritiated ligands in radioreceptor crosslinking, particularly those of low molecular weight which might otherwise undergo steric modification due to iodination, this having the potential for interfering with receptor ligand binding.     

The N-Terminal End Truncated Mu-Opioid Receptor: from Expression to Circular Dichroism Analysis

In order to evaluate the biochemical, biophysical, and pharmacological implication of the N-terminal domain of the human mu-opioid receptor (HuMOR), deletion mutants lacking 64 amino acids from the amino terminus of HuMOR were constructed and expressed in the yeast Pichia pastoris. The recombinant proteins differed with respect to the presence of the Saccharomyces cerevisiae α-factor prepropeptide and the enhanced green fluorescent protein fused to the N terminus of the receptor. Pharmacological studies indicated that deletion of the N-terminal domain produced little effect on ligand affinities. The N-terminal end truncated and c-myc/6his-tagged receptor was subsequently purified to homogeneity and a yield of 5 mg/l was obtained after purification. The N-terminal end truncated receptor was further characterized by circular dichroism in trifluoroethanol and showed a characteristic pattern of α-helical structure. A pH effect on the structure of the receptor was observed when it was solubilized in sodium dodecyl sulfate micelles, with an increase of helicity at low pH.     

Affinity chromatography in purification of A1 adenosine receptors.

Purification of A1 adenosine receptor of rat brain membranes was performed using a newly developed affinity gel employing xanthine amine congener (XAC) as an immobilized ligand. The A1 adenosine receptor was solubilized with digitonin-cholate from brain membranes and then purified by a sequential use of affinity chromatography on XAC-agarose, hydroxyapatite chromatography and reaffinity chromatography on XAC-agarose. The A1 adenosine receptor was purified ca. 45,000-fold with a yield of 5%. The final receptor preparation gave a single broad band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a Mr approximately 34,000. This band was also shown to be specifically labelled with an affinity labelling reagent for A1 adenosine receptors. This purification method was also applicable for the complete purification of A1 adenosine receptors from rat testis and human brain membranes.     

LCZ696: a dual-acting sodium supramolecular complex

LCZ696 displays several surprising features, such as an intrinsic stabilization of the supramolecular complex by monovalent multi-coordinated sodium cations and water molecules. In addition, to the best of our knowledge, the compound is the first example of a dual-acting pharmaceutical built as a supramolecular complex delivering two pharmacologic effects—angiotensin receptor 1 (AT1) blockage and neprilysin (NEP) inhibition.     

Bioinspired Artificial Nanodecoys for Hepatitis B Virus

A facile route is presented for fabricating a new class of nanomimics that overexpress hepatitis B virus (HBV) receptor by a natural biosynthetic procedure against HBV infection. A nine‐transmembrane HBV‐specific receptor, human sodium taurocholate co‐transporting polypeptide (hNTCP), was engineered to naturally immobilize it onto the cellular surface and subsequently trigger the budding of hNTCP‐anchoring membrane vesicles (hNTCP‐MVs) that favor the HBV virion. hNTCP‐MVs could rapidly block HBV infection in cell models. Furthermore, hNTCP‐MVs treatment could effectively prevent viral infection, spreading, and replication in a human‐liver‐chimeric mouse model of HBV infection. Our findings demonstrate the receptor‐mediated antiviral effect of hNTCP‐MVs to trick HBV and offer novel opportunities for further development of antiviral strategies in nanomedicine.