Molecular umbrella-assisted transport of glutathione across a phospholipid membrane

A di-walled molecular umbrella (1a) has been synthesized by acylation of the terminal amino groups of spermidine with cholic acid, followed by condensation with bis(3-O-[N-1,2,3-benzotriazin-4(3H)-one]yl)-5,5'-dithiobis-2-nitrobenzoate (BDTNB), and displacement with glutathione (gamma-Glu-Cys-Gly, GSH). Replacement of the sterol hydroxyls with sulfate groups, prior to displacement with GSH, afforded a hexasulfate analogue 1b. Both conjugates have been found to enter large unilamellar vesicles (200 nm diameter, extrusion) of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and to react with entrapped GSH to form oxidized glutathione (GSSG). Evidence for vesicular entry has come from the formation of oxidized glutathione (GSSG) within the interior of the vesicle, the appearance of the thiol form of the umbrella (USH), and the absence of release of GSH into the external aqueous phase. Results that have been obtained from monolayer experiments, together with the fact that the heavily sulfated conjugate is able to cross the phospholipid bilayer, have yielded strong inferential evidence for an "umbrella-like" action of these molecules as they cross the lipid bilayer.     

Nitrate and chloride transport through a smart membrane

To develop membranes having ionic selective properties under control of external stimuli is a challenge of the membrane and material scientific community. Conducting polymers swell and shrink under electrochemical control, so they are good candidates to prepare such smart membranes. The ionic transport through a new free-standing polypyrrole film working as a membrane in a diffusion cell was studied. The driving forces were transversal electric fields or concentration gradients across the film. The obtained ionic conductivity was dependent on both the electrolyte nature and concentration, as well as on the oxidation degree of the film, which was controlled by the applied external electric potential. Reverse and continuous changes of up to one order of magnitude on the transversal ionic conductivity are obtained when the membrane is in stationary oxidation states attained by polarisations at a constant potential in the range between −0.6 V and +0.4 V, respectively. A prevalent conductivity of anions (t₋ = 0.94) was obtained from Donnan potential measurements. The experimental results indicate that the oxidised film behaves as a nanoporous membrane highly permeable to nitrate ions, while the rejection of these ions is very high in the reduced film. The free-standing polypyrrole film works then as a smart membrane selective to nitrate ions under concentration gradient.     

Supported phospholipid membrane interactions with 1-butyl-3-methylimidazolium chloride

Quartz crystal microbalance with dissipation measurements were conducted for a 98/2 mole ratio of 1,2-dielaidoylphosphocholine (DEPC) and 1,2-dimyristoylphosphoglycerol (DMPG) on silica, gold, and a self-assembled monolayer of 11-mercapto-1-undecanol (11MU) and 11-mercaptoundecanoic acid (11MUA) at 50 mol % each. This study demonstrates that vesicles composed of DEPC and DMPG at 98 and 2 mol %, respectively, formed a supported bilayer with unruptured vesicles present when adsorbed onto the self-assembled monolayer. Also, the partially formed supported bilayer apparently deadsorbed in the presence of 1-butyl-3-methylimidazolium chloride, suggesting that surface-bilayer interactions are weaker on a hydrophilic modified gold surface composed of 50/50 11MU/11MUA than the surface-bilayer interactions on silica.     

Structure-activity relationships in cholapod anion carriers: enhanced transmembrane chloride transport through substituent tuning

Chloride transport by a series of steroid-based "cholapod" receptors/carriers was studied in vesicles. The principal method involved preincorporation of the cholapods in the vesicle membranes, and the use of lucigenin fluorescence quenching to detect inward-transported Cl-. The results showed a partial correlation between anion affinity and transport activity, in that changes at the steroidal 7 and 12 positions affected both properties in concert. However, changes at the steroidal 3-position yielded irregular effects. Among the new steroids investigated the bis-p-nitrophenylthiourea 3 showed unprecedented activity, giving measurable transport through membranes with a transporter/lipid ratio of 1:250 000 (an average of <2 transporter molecules per vesicle). Increasing transporter lipophilicity had no effect, and positively charged steroids had low activity. The p-nitrophenyl monourea 25 showed modest but significant activity. Measurements using a second method, requiring the addition of transporters to preformed vesicle suspensions, implied that transporter delivery was problematic in some cases. A series of measurements employing membranes of different thicknesses provided further evidence that the cholapods act as mobile anion carriers.     

Molecular umbrella-assisted transport of an oligonucleotide across cholesterol-rich phospholipid bilayers

A series of molecular umbrella conjugates, derived from cholic acid, deoxycholic acid, spermidine, lysine, and 5-mercapto-2-nitrobenzoic acid, have been synthesized and found capable of transporting an attached 16-mer oligonucleotide (S-dT16) across liposomal membranes made from 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidyldglycerol (POPG), and cholesterol [POPC/POPG/cholesterol (65/5/30; mol/mol/mol, v/v/v)] at 37 degrees C. Those molecular umbrellas containing four choloyl (or deoxycholoyl) groups resulted in significantly faster rates of transport as compared to those containing only two such moieties. A model that accounts for these membrane transport processes is proposed.     

Electric mass transport of sodium chloride through cation-exchange membrane MK-40 in dilute sodium chloride solutions: A rotating membrane disk study

The polarization properties of an electromembrane system consisting of an MK-40 membrane and a dilute sodium chloride solution are investigated with an experimental apparatus, which includes a rotating membrane disk with a horizontally positioned membrane. For the electrochemical systems of MK-40/0.01 M NaCl and MK-40/0.001 M NaCl, effective ion transport numbers and partial current-voltage curves are determined for sodium and hydrogen ions, and limiting-current densities and the diffusion-layer thickness are calculated as functions of the rotation rate of the membrane disk. The space-charge distribution in the diffusion layer and in the membrane is calculated for various current densities and rotation rates of the membrane. It is shown that when electric-current densities are greater than the limiting value, ion fluxes of the salt increase as a result of a decrease in the effective thickness of the diffusion layer. This decrease is caused by the development of space charge, electroconvection, water dissociation, and the exaltation effect in the region near the membrane. It has been established that in dilute solutions the limiting current is not purely electrodiffusive in nature.     

A needle-and-thread approach to bilayer transport: permeation of a molecular umbrella-oligonucleotide conjugate across a phospholipid membrane

A di-walled molecular umbrella, composed of two choloyl groups, one spermidine moiety, and a 5-thiol(2-nitrobenzoyl) "handle", was covalently attached to a 16-mer oligonucleotide (S-dT16) through a disulfide bond. Incubation of this conjugate (1) with vesicles made from 1-palmitoyl-2-oleyol-sn-glycero-3-phosphocholine and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (95/5, mol/mol) (200 nm diameter, extrusion) that contained entrapped glutathione (GSH) at 37 degrees C resulted in the liberation of the oligonucleotide and the umbrella-bound 5-mercapto(2-nitrobenzoyl) moiety (USH) via thiolate-disulfide interchange. The appearance of USH, together with the absence of leakage of entrapped GSH and a vesicular capture of the oligonucleotide that matches the extent of USH formation, provides compelling evidence for delivery of S-dT16 into the aqueous compartment of these vesicles. In a sense, the molecular umbrella functions like a "needle" in providing a pathway for the oligonucleotide (the "thread") to cross the membrane.     

A synthetic, chloride-selective channel that alters chloride transport in epithelial cells

An Ussing chamber was used to demonstrate that synthetic amphiphilic anion transporters function as chloride transporters in mammalian airway epithelial cells.     

Effect of the structure of functionalized phosphoryl carriers on the membrane transport of proton-donor substrates

A series of phosphoryl compounds functionalized in the side chain were synthesized, and their membrane-transport properties with respect to proton-donor substrates of various acidity were studied. It was found that the efficiency of phase transport of the strong monobasic perchloric acid correlates with the basicity of the phosphoryl carriers in a series of carriers containing oxygen-containing functional groups. The transport flow sharply increases in going to phosphorylated amines, whereas phosphoramidates in their efficiency are closer to phosphonates than to amines. The efficiency of transport of dibasic acids (oxalic and tartaric) is low, since the hydroxy and carboxy groups not bound to the carrier make ionic associates highly hydrophilic. Fine details of the structure-transport acitivity relationship in the series of phosphorus compounds were discussed. Three-dimensional correlation analysis was used to compare the structure of the carriers with their characteristics: basicity of amine centers, atomic charges of oxygen and nitrogen, and hardness and hydrophobicity parameters.     

Aggregation and supramolecular membrane interactions that influence anion transport in tryptophan-containing synthetic peptides

Self-assembly is a desired property in supramolecular chemistry, but extensive aggregation may be counterproductive. Rigid systems typically have better organization, but are inherently less dynamic. This work shows that ion transport by amphiphilic heptapeptides (synthetic anion transporters or SATs) is affected by aggregation of the monomers in the bulk aqueous phase to which they are added and within the bilayer. Ion transport was assessed for all compounds by assay of Cl(-) release from liposomes. The mechanism of ion transport was confirmed by planar bilayer conductance studies for two compounds at opposite ends of the efficacy scale. Dynamic light scattering, the Langmuir trough, transmission electron microscopy, ion release from liposomes, and planar bilayer conductance studies were used to assess the importance of self-assembly versus aggregation in ion transport. Generally, greater aggregation was has an adverse effect on the transport, although at least dimerization is required for amphiphilic heptapeptides to readily transport Cl(-). Anion transport in these systems was found to be sensitive to changes in the C-terminal portion of the (Gly)(3)Pro(Gly)(3) sequence. Moreover, a significant difference in transport efficacy was apparent when L-Trp was replaced by D-Trp in the same position.     

Synergetic transport of europium through a contained supported liquid membrane using trioctylamine and tributyl phosphate as carriers

Contained supported liquid membrane (CSLM) system has been explored for synergic liquid membranes. Different experimental variables such as concentration of acid, carrier, metal ion and complexing agents, stirring speed, etc. have been investigated. Trioctylamine (TOA) + tributyl phosphate (TBP) system has been explored for europium.     

Solvent-polymer interaction: I. Molecular transport of some selected organic liquids in polymer membrane

Methods and microtechniques for determining solubility, diffusivity, thermodynamic properties, and kinetic parameters of 12 selected organic liquid solvents in polyurethane membrane by thermogravimetry (TG) are described. TG provides a simple, sensitive, rapid, and accurate microtechnique for measuring a minute change in weight (or mass) of a substance as a function of time at isothermal condition or as a function of temperature at dynamic manner. Thus from a single isothermal TG-desorption experiment, solubility and diffusivity of solvent molecules in polyurethane membrane were obtained simultaneously. Furthermore, by a dynamic TG-desorption run, kinetic parameter such as activation energy of desorption of solvent molecules from polyurethane membrane was determined. In addition, much other useful information such as equilibrium sorption constant, the changes in standard enthalpy and standard entropy of sorption, permeability, the activation energy of diffusion and so forth for solvents in polyurethane membrane are also evaluated and discussed. Finally, the correlation between the microscopic molecular structure and macroscopic properties of solvent molecules in polymer membrane is interpreted in terms of linear free energy relationships.     

Wet poly(vinyl chloride) membrane as a support: Sorption and transport of low-molecular-weight organic compounds and proteins

Characteristics of sorption and transport behavior of “wet” (PVC) membranes prepared by a poly(vinyl chloride) casting method were studied. It was found that wet PVC membranes adsorbed enzymes and proteins, whilst they did not adsorb low-molecular-weight compounds. The diffusivity of acetylcholine iodide as a penetrant through the wet PVC membrane was approximately 10^?7 cm²/sec, although it depended strongly on membrane thickness and slightly on PVC concentrations in the casting solution. Activation energy of acetylcholine iodide diffusivity was 2.6 kcal/mol. These results were explained by hydrophobic interaction and minute pore dimensions. Wet PVC membrane was suggested as a support for immobilization and was compared with collagen on the basis of transport behavior.     

Generation and transport of charge carriers in polyvinylcarbazole

The processes of generation and transport of excess charge carriers in polyvinylcarbazole at room temperature and at 353 K were experimentally studied. The polymer was charged with pulses of electrons at energies of 7 and 50 keV, which differed in both linear energy transfer and track structure. A universal method of investigation based on the combination of the time-of-flight technique in the both (surface and bulk) modes with the measurement of radiation-induced conductivity was used. The radiation-chemical yield of free charges was measured using two independent procedures. It was shown that the radiation chemical yield of free charges at 293 K was somewhat smaller for 7-keV than for 50-keV electrons; in the latter case, G = 1.1 in an electric field of 2 × 10⁷ V/m. The parameters of the generalized physical model were determined for polyvinylcarbazole.     

Photogeneration and transport of charge carriers in polysilylenes

The attachment of π-conjugated chromophores that absorb the radiation with long wavelengths to poly(methylphenylsilylene) ( 1 ) via reactions of its formylated derivative is described. Some of the polymers obtained show improved photostability and higher quantum photogeneration efficiency in comparison with the parent polymer. Photoconductive ultra-thin layers can be prepared from polar derivatives of ( 1 ) by the Langmuir–Blodgett technique.     

Supramolecular membrane transport: From biomimics to membrane sensors

A stable, long-lived membrane sensor for dissolved oxygen is reported. A conventional amperometric Clark cell was augmented through the addition of an ion-exchange carrier to the membrane to permit export of hydroxide in exchange for chloride in the sample solution. The choice of a suitable carrier was determined from two types of supramolecular principles: (1) the characteristic flux as a function of the magnitude of the two-phase ion exchange equilibrium constant (K_ex) for an antiport transport cycle as derived for biomimetic ion transport studies and (2) the use of guanidinium ion exchangers to provide hydrogen-bonding in addition to electrostatic recognition for enhanced hydroxide/chloride selectivity. The membrane of the sensor supports the anticipated ion-exchange as it continues to provide stable current beyond the point where the initial internal chloride would be entirely consumed. As a consequence, stable and responsive sensors can be fabricated using planar techniques such as screen printing.     

Liquid membrane transport: a survey

The literature pertaining to facilitated transport and liquid membrane separations is reviewed and summarized, especially work reported since 1977. Liquid membranes of all geometries are discussed, including immobilized liquid membranes and liquid surfactant or emulsion liquid membranes. Emphasis is placed on facilitated, or carrier-mediated transport in both configurations although other mechanisms such as coupled-transport and transport due to solubility differences are discussed. Mathematical modeling and analytical solutions for facilitated transport models are summarized. The possibility of industrial application of liquid membrane technology is mentioned and the most important experimental techniques for liquid membrane research are discussed. Also, directions for future research are recommended.     

New fluorescent probes reveal that flippase-mediated flip-flop of phosphatidylinositol across the endoplasmic reticulum membrane does not depend on the stereochemistry of the lipid

Glycerophospholipid flip-flop across biogenic membranes such as the endoplasmic reticulum (ER) is a fundamental feature of membrane biogenesis. Flip-flop requires the activity of specific membrane proteins called flippases. These proteins have yet to be identified in biogenic membranes and the molecular basis of their action is unknown. It is generally believed that flippase-facilitated glycerophospholipid flip-flop across the ER is governed by the stereochemistry of the glycerolipid, but this important issue has not been resolved. Here we investigate whether the ER flippase stereochemically recognizes the glycerophospholipids that it transports. To address this question we selected phosphatidylinositol (PI), a biologically important molecule with chiral centres in both its myo-inositol headgroup and its glycerol-lipid tail. The flip-flop of PI across the ER has not been previously reported. We synthesized fluorescence-labeled forms of all four diastereoisomers of PI and evaluated their flipping in rat liver ER vesicles, as well as in flippase-containing proteoliposomes reconstituted from a detergent extract of ER. Our results show that the flippase is able to translocate all four PI isomers and that both glycerol isomers of PI flip-flop across the ER membrane at rates similar to that measured for fluorescence-labeled phosphatidylcholine. Our data have important implications for recent hypotheses concerning the evolution of distinct homochiral glycerophospholipid membranes during the speciation of archaea and bacteria/eukarya from a common cellular ancestor.     

Stabilized phospholipid membranes in chromatography: toward membrane protein-functionalized stationary phases

Transmembrane protein (TMP)-functionalized materials have resulted in powerful new methods in chemical analysis. Of particular interest is the development of high-throughput, TMP-functionalized stationary phases for affinity chromatography of complex mixtures of analytes. Several natural and synthetic phospholipids and lipid mimics have been used for TMP reconstitution, although the resulting membranes often lack the requisite chemical and temporal stability for long-term use, a problem that is exacerbated in flowing separation systems. Polymerizable lipids with markedly increased membrane stability and TMP functionality have been developed over the past two decades. More recently, these lipids have been incorporated into a range of analytical methods, including separation techniques, and are now poised to have a significant impact on TMP-based separations. Here, we describe current methods for preparing TMP-containing stationary phases and examine the potential utility of polymerizable lipids in TMP affinity chromatography.     

The synthesis and membrane transport characteristics of macrocyclic polyether ligands composed of 1,10-phenanthroline as carriers for primary amine species

Four macrocyclic polyether derivatives of o-phenanthroline were synthesized and used as neutral carriers for preparing poly(vinyl chloride) (PVC) membrane electrodes to sense primary amine species. The potentiometric response characteristics of the electrodes prepared were investigated. The electrode sensitive to benzyl amine as model analyte showed a linear response range of 8.0 x 10(-6)-0.1 mol/l with a detection limit of 8.9 x 10(-7) mol/l and a slope of 56.5 mV/decade. The linear potentiometric response of the mexiletine-sensitive electrode was 4.7 x 10(-6)-0.1 mol/l, and the detection limit was 5.0 x 10(-7)1 mol/l with a slope of 59.0 mV/decade. The transfer behavior of amines and ammonium ions through an organic phase was investigated by means of the bulk liquid membrane transport experiment. The effects of pH, counter anions and other factors on the transfer of the amine and ammonium species were studied. The mass transfer rates of the test species facilitated by macrocyclic polyether derivatives of o-phenanthroline were determined and the following sequence was found: benzyl amine > ethyl amine > tetramethyl ammonium > triethyl amine > diethyl amine > K(+) > ammonium > Na(+) > Ca(2+) > Mg(2+). This was exactly the potentiometric selectivity sequence of the membrane electrodes prepared by using these carriers. The mechanism of transfer of benzyl amine through a membrane phase induced by the carriers has been elucidated on the basis of experimental observations.