Structural and mechanistic motifs in membrane proteins: the three-dimensional modelling of rhodopsin, band 3, and the nicotinic acetylcholine receptor

Because so little is known about the structure of membrane proteins, an attempt has been made in this work to develop techniques by which to model them in three dimensions. The procedures devised rely heavily upon the availability of several sequences of a given protein. The modelling procedure is composed of two parts. The first identifies transmembrane regions within the protein sequence on the basis of hydrophobicity, β-turn potential, and the presence of certain amino acid types, specifically, proline and basic residues. The second part of the procedure arranges these transmembrane helices within the bilayer based upon the evolutionary conservation of their residues. Conserved residues are oriented toward other helices and variable residues are positioned to face the surrounding lipids. Available structural information concerning the protein's helical arrangement, including the lengths of interhelical loops, is also taken into account. Rhodopsin, band 3, and the nicotinic acetylcholine receptor have all been modelled using this methodology, and mechanisms of action could be proposed based upon the resulting structures.

Specific residues in the rhodopsin and iodopsin sequences were identified, which may regulate the proteins' wavelength selectivities. A hinge-like motion of helices M3, M4, and M5 with respect to the rest of the protein was proposed to result in the activation of transducin, the G-protein associated with rhodopsin. A similar mechanism is also proposed for signal transduction by the muscarinic acetylcholine and β-adrenergic receptors.

The nicotinic acetylcholine receptor was modelled with four trans-membrane helices per subunit and with the five homologous M2 helices forming the cation channel. Putative channel-lining residues were identified and a mechanism of channel-opening based upon the concerted, tangential rotation of the M2 helices was proposed.

Band 3, the anion exchange protein found in the erythrocyte membrane, was modelled with 14 transmembrane helices. In general the pathway of anion transport can be viewed as a channel composed of six helices that contains a single hydrophobic restriction. This hydrophobic region will not allow the passage of charged species, unless they are part of an ion-pair. An arginine residue located near this restriction is proposed to be responsible for anion transport. When ion-paired with a transportable anion it rotates across the barrier and releases the anion on the other side of the membrane. A similar process returns it to its original position. This proposed mechanism, based on the three-dimensional model, can account for the passive, electroneutral, anion exchange observed for band 3. Dianions can be transported through a similar mechanism with the additional participation of a histidine residue. Both residues are located on M10.

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

This study examined the toxic effects of microcystins on mitochondria of liver and heart of rabbit in vivo. Rabbits were injected i.p. with extracted microcystins (mainly MC-RR and -LR) at two doses, 12.5 and 50 MCLReq. mu g/kg bw, and the changes in mitochondria of liver and heart were studied at 1, 3,12, 24 and 48 h after injection. MCs induced damage of mitochondrial morphology and lipid peroxidation in both liver and heart. MCs influenced respiratory activity through inhibiting NADH dehydrogenase and enhancing succinate dehydrogenase (SDH). MCs altered Na+-K+-ATPase and Ca2+-Mg2+-ATPase activities of mitochondria and consequently disrupted ionic homeostasis, which might be partly responsible for the loss of mitochondrial membrane potential (MMP). MCs were highly toxic to mitochondria with more serious damage in liver than in heart. Damage of mitochondria showed reduction at 48 h in the low dose group, suggesting that the low dose of MCs might have stimulated a compensatory response in the rabbits. (C) 2008 Elsevier Inc. All rights reserved.

Proteomic analysis of the sarcosine-insoluble outer membrane fraction of Flavobacterium columnare

Outer membrane proteins (OMPs) of bacteria are key molecules interacting with the host environment. Flavobacterium columnare, a pathogen-causing columnaris disease of fish worldwide, was studied in order to understand the composition of its OMPs. The sarcosine-insoluble membrane fraction of the OMPs was analysed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with reverse-phase high-performance liquid chromatography-tandem mass spectrometry (RP-HPLC MS/MS). Thirty-six proteins were identified, including proteins involved in cell wall/membrane biogenesis, specific transport of various nutrients and in essential metabolism. The present study is the first report on the OMPs of F. columnare, and may serve as the basis for understanding the pathogenesis of the bacterium.

Guiding of 150 keV O6+ ions through nanocapillaries in an uncoated Al2O3 membrane: special time dependence of the transmission profile width

This paper reports that the transmission of O6+ ions with energy of 150keV through capillaries in an uncoated Al2O3 membrane was measured, and agreements with previously reported results in general angular distribution of the transmitted ions and the transmission fractions as a function of the tilt angle well fitted to Gaussian-like functions were observed. Due to using an uncoated capillary membrane, our c is larger than that using a gold-coated one with a smaller value of E-p/q, which suggests a larger equilibrium charge Q(infinity) in our experiment. The observed special width variation with time and a larger width than that using a smaller E-p/q were qualitatively explained by using mean-field classical transport theory based on a classical-trajectory Monte Carlo simulation.

Blends based on sulfonated poly[bis(benzimidazobenzisoquinolinones)] and poly(vinylidene fluoride) for polymer electrolyte membrane fuel cell

A new blend system consisting of an amorphous sulfonated poly[bis(benzimidazobenzisoquinolinones)] (SPBIBI) and the semi-crystalline poly(vinylidene fluoride) (PVDF) was prepared for proton exchange membranes. The miscibility behavior of a series of blends of SPBIBI with PVDF at various weight ratios was studied by WXRD, DSC and FTIR. The properties of the blend membranes were investigated, and it was found that the introduction of PVDF in the SPBIBI matrix altered the morphological structure of the blend membranes, which led to the formation of improved connectivity channels. For instance, the conductivity of the blend membrane containing 10 wt% PVDF displayed the highest proton conductivity (i.e., 0.086 S cm(-1)) at room temperature, a value almost twofold that of the pristine SPBIBI membranes (i.e., 0.054S cm(-1)) under identical conditions.

Synthesis and characterization of soluble poly(amideimide)s bearing triethylamine sulfonate groups as gas dehumidification membrane material

A series of soluble poly(amide-imide)s (PAIs) bearing triethylammonium sulfonate groups were synthesized directly using trimellitic anhydride chloride (TMAC) polycondensation with sulfonated diamine such as 2,2'-benzidinedisulfonic acid (BDSA), 4,4'-diaminodiphenyl ether-2,2'-disulfonic acid (ODADS), and nonsulfonated diamine 4,4-diaminodiphenyl methane in the presence of triethylamine. The resulting copolymers exhibited high molecular weights (high inherent viscosity), and a combination of desirable properties such as good solubility in dipolar aprotic solvents, film-forming capability, and good mechanical properties. Wide-angle X-ray diffraction revealed that the polymers were amorphous. These copolymers showed high permeability coefficients of water vapor because of the presence of the hydrophilic triethylammonium sulfonate groups. The water vapor permeability coefficients (P-w) and permselectivity coefficients of water vapor to nitrogen and methane [alpha(H2O/N-2) and (alpha(H2O/CH4)] Of the films increased with increasing the amount of the triethylammonium sulfonated groups.

A new flow field and its two-dimension model for polymer electrolyte membrane fuel cells (PEMFCs)

A new flow field was designed to search flow fields fitting polymer electrolyte membrane fuel cells (PEMFCs) better due its extensible. There are many independent inlets and outlets in the new flow field. The new flow field we named NINO can extend to be more general when pressures at the inlet and outlet vary and some usual flow fields will be obtained. A new mathematical model whose view angle is obverse is used to describe the flow field.

A new kind of potassium sensor based on capacitance measurement of mimic membrane

A novel type of potassium sensor based on the capacitance change of valinomycin-incorporated bilayer supported on a gold electrode has been developed and characterized. The lipid membrane was Formed by painted method and monitored simultaneously by capacitance variation. The capacitance of the electrode-supported membrane was found to be modulated by different concentrations of K+. Investigating the capacitance change allows a simple and specific technique for the measurement of potassium ion in solution. Especially, the homemade capacitance meter is, to our knowledge, used to monitor the bilayer membrane formation and detect K+ for the first time. It has been proved that this capacitance measurement is a very useful technique because it is simple and sensitive compared to the other methods.

Proton transfer across conducting polypyrrole membrane separated by two electrolyte solutions

Electrochemical behavior of the transfer of H+ across polypyrrole membrane (PPM) was studied. The transfer process was quasi-reversible and mainly diffusion-controlled. PPM electropolymerized in water solution has better reversibility than that in CH3CN solution for the transfer of H+. The transfer process of H+ across the two kinds of PPM indicated that the PPM electrochemically polymerized was of asymmetry.

Permeation of nitrogen and water vapor through sulfonated polyetherethersulfone membrane

The novel polyetherethersulfone (PES-C) prepared from phenol-phthalein in our institute is an amorphous, rigid, tough material with good mechanical properties over a wide temperature range. To improve its water vapor permeability for the application of gas drying, the PES-C was sulfonated with concentrated sulfuric acid and transferred in sodium, cupric, and ferric salt forms. The sulfonation degree can be regulated by controlling the temperature and reaction time. Characterization of sulfonated PES-C in sodium form was made by IR. Some properties of the sulfonated PES-C, such as solubility, glass transition temperature, thermal stability, mechanical properties, and transport properties to nitrogen and water vapor have also been discussed. (C) 1997 John Wiley & Sons, Inc.

Novel poly(aryl ether ketone)s containing various pendant groups .2. Gas-transport properties

The gas transport of hydrogen, oxygen, nitrogen, carbon dioxide, and methane gases in a series of poly(aryl ether ketone)s was examined. These polymer membranes have a wide range of permeability coefficients and permselectivity coefficients, showing excellent gas-transport properties. The enhanced interchain interaction in the polymers due to intermolecular hydrogen bonds and ionic bonds results in a considerable increase in permselectivity but a decrease in permeability. On the contrary, the polymers with bulky arkyl substituents show significantly increased permeability. The causes of this trend are interpreted in terms of the free volume, interchain distance, and glass transition temperature together with the respective contribution of gas solubility and diffusivity to the overall permeability. Of interest is the observation that the ionomer IMPEK-K+, which simultaneously contains bulky isopropyl substituents and pendant carboxylate groups, exhibits over twice higher CO2 permeability and 15% higher CO2/CH4 permselectivity than those of bisphenol-A p'olysulfone (PSF). The possibility of using the new synthesized poly(aryl ether ketone)s in gas separation membrane application is also discussed. (C) 1997 John Wiley & Sons, Inc.

Electrochemical studies of lipophilic ion transport through BLM. The influence of sterols on its transport

Cyclic voltammetry was employed to study the influence of sterols on the lipophilic ion transport through the BLM. The mole fraction of the sterols (cholesterol, oxidized cholesterol). as referred to total lipid, was varied in a range of 0-0.8. Data demonstrate that a thin-layer model is suitable to this BLM system. By this model, the number of charges transported per lipophilic ion, the concentration of the ion in the membrane bulk phase and the aqueous/membrane phase partition coefficient could be calculated. These parameters proved that sterols had an obvious influence on the lipophilic ion transport. Cholesterol had a stronger influence on the ion transport than oxidized cholesterol. Its incorporation into egg lecithin membranes increased the partition coefficient beta of the ion up to more than 3-fold. Yet, oxidized cholesterol incorporated into egg lecithin membranes only increased the beta up to less than 2-fold, and the beta had no great variation at different oxidized cholesterol mole fractions. The higher beta obtained was partly due to the trace amount of solvent existing in the core of the lipid bilayers. At the different sterol mole fractions, combining the change of beta with the change of peak current, we also concluded that sterols had somewhat inhibiting effect on the ion transport at the higher sterols mole fraction (>0.4). These results are explained in terms of the possible change of dipole potential of the membrane produced by sterols and the decrease of the membrane fluidity caused by the condensation effect of sterols and the thinning effect caused by sterols. The substituting group (in the oxidized cholesterol) had some inhibiting effects on the ion transport at higher mole fractions (oxidized cholesterol mole fraction >0.4).

The transfer of chloride ion across an anion exchange membrane

The transfer of chloride ions into a low resistance anion exchange membrane (AEM) was investigated by cyclic voltammetry (CV) and electrochemical impedance spectra. In all cases, concentration polarization of Cl- ions is exterior to the membrane. It controls the flux and produces the limiting currents: either steady state or transient (peak type) current. In CV experiments, when the size of the holes in the membrane was much smaller than the distance between membrane holes, the Cl- anion transfer showed steady state voltammetric behavior. Each hole in the membrane can be regarded as a microelectrode and the membrane was equivalent to a microelectrode array in this condition. When the hole in the membrane was large or the distance between membrane holes was small, the CV curve of the Cl- anion transfer across the membrane showed a peak shape, which was attributed to linear diffusion. In AC impedance measurement, the impedance spectrum of the membrane system was composed of two semicircles at low DC bias, corresponding to the bulk characteristics of the membrane and the kinetic process of ion transfer, respectively. The bulk membrane resistance increases with increasing DC bias and only one semicircle was observed at higher DC bias. The parameters related to kinetic and membrane properties were discussed.

STUDY ON MASS-TRANSPORT IN EASTMAN-AQ POLYMER FILM BY USING NON-STEADY-STATE CHRONOAMPEROMETRY AT AN ULTRAMICRODISK ELECTRODE

Non-steady-state chronoamperometry of ultramicroelectrodes is a powerful method for the study of mass transport in polymer films. This method has many advantages over the conventional methods at a macroelectrode and the steady state method at an ultramicroelectrode, which yield the most information. The apparent diffusion coefficient, D(app), and the concentration of reactant in the film, c(f), can be determined from a single experiment without knowing the thickness of the film. We studied the transport of several species such as Ru(NH3)63+, Ru(bpy)3(2+), NR and MV2+ in Eastman-AQ polymer film coated ultramicroelectrodes by using this method.

AN EXPERIMENTAL-STUDY ON MEMBRANE DISTILLATION FOR TREATMENT OF GINSENG DEW AND GINSENG WASHING WATER

Membrane distillation is a new membrane separation process which has been developed in the last few years. When a piece of microporous hydrophobic membrane separates two kinds of aqueous solutions different in temperature, the solutions cannot transport through the pores of membrane in any directions because of the hydrophobicity of membrane. However, vapor can readily penetrate through the