Functional immobilisation of the nicotinic acetylcholine receptor in tethered lipid membranes

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We have examined the properties of nicotinic acetylcholine receptors in embryonic chick retinal ganglion cells. Ganglion cells, identified according to morphological and physiological criteria, displayed spontaneous or induced action potentials. In 94/99 cells acetylcholine pulses evoked responses. In current clamp mode, acetylcholine provoked membrane depolarization and triggered action potentials. Under voltage clamp conditions, acetylcholine evoked inward currents that were readily blocked by d-tubocurar

The GABA-synthesizing enzyme GAD65 is synthesized as a soluble cytosolic protein but undergoes post-translational modification(s) to become anchored to the cytosolic face of Golgi membranes before targeting to synaptic vesicle membranes in neuroendocrine cells. Palmitoylation of cysteines 30 and 45 in GAD65 is not required for targeting to Golgi membranes but is crucial for post-Golgi trafficking to presynaptic clusters in neurons. Here, we show that palmitoylated GAD65 colocalizes with the small GTP- bindi

A new type of Nafion/Fe structured membrane ensuring faster kinetics, higher efficiency, and mechanical properties has been prepared and will be compared in its performance with the Fe-exchanged commercial Dupont 117 Nafion/Fe membrane during the abatement of model organic compounds. During the casting of the laboratory Nafion sample, the iron ions were introduced directly into the Nafion oligomer solution. This novel laboratory Nafion/Fe was tested as an immobilized catalyst in the degradation of several t

High signal-to-noise Fourier transform IR (FTIR) spectra of the 5-hydroxytryptamine (serotonin) receptor (5-HT3R) and the nicotinic acetylcholine receptor (nAChR) were obtained by microscope FTIR spectroscopy using micrometer-sized, fully hydrated protein films. Because this novel procedure requires only nanogram quantities of membrane proteins, which is 4-5 orders of magnitude less than the amt. of protein typically used for conventional FTIR spectroscopy, it opens the possibility to access the structure a

Trinuclear metallamacrocycles can be obtained by reactions of (arene)Ru, CpRh and CpIr complexes with pyridonate ligands. Using 3-oxy-pyridonate as the bridging ligand, organometallic analogues of 12-crown-3 are formed, which display an extremely high affinity and selectivity for lithium and sodium ions. In the presence of guest molecules, the redox-potential of the metallamacrocyclic hosts is shifted by more than 300 mV towards positive potential suggesting potential applications as chemosensors. The aff

Accurate online monitoring of biomass is an enduring problem. Of the various methods available, one of the more satisfactory is with dielec. spectroscopy (often referred to as capacitance monitoring). However, as this technique measures total -biovolume', large changes in cellular morphol. or population distribution will alter the calibration with respect to cell wt. or no. In an attempt to minimize this problem, the full dielec. spectrum was scanned every 10-20 min, and then multivariate techniques were us

Polyelectrolyte complex membrane formation has been studied, online, using the synthetic boundary technique of an Optima XL-I anal. ultracentrifuge. This recent method, established by the first author, has now been evaluated concerning its sensitivity to monitor differences in the membrane formation if the membrane components and exptl. conditions are varied. For this purpose the membrane formation of sodium alginate with three polycations, poly(vinylamine)hydrochloride, poly(L-lysine)hydrochloride, and chi

The detn. of residual dipolar couplings (RDCs) by quant. J spectroscopy methods such as heteronuclear single quantum correlation with phase encoded coupling (HSQC-PEC) is prone to systematic errors that may be caused by differential attenuation during the conversion of orthogonal d. operator components into observable terms. The attenuation may be caused by miscalibration of r.f. pulses and by relaxation effects. A simple method is presented that allows one to remove most of these systematic errors without

Fluorescence Correlation Spectroscopy (FCS) is a suitable technique to investigate biomol. interactions. The advantage of FCS lies in its wide dynamic range in the time and concn. domain, its sensitivity, and thus in the small amt. of material needed. The characteristic time of the processes that can be measured spans more than eleven orders of magnitude, between several nanoseconds up to hours; the measurable concn. range extends from 0.1 nM to several mM. Here we report on FCS expts. to investigate the pr

The binding of the fluorescein-labeled antagonist GR-flu ([1,2,3,9-tetrahydro-3-[(5-methyl-1H-imidazol-4-yl)methyl]-9-(3-amino- (N-fluoresceinthiocarbamoyl)propyl)-4H-carbazol-4-one]) to a purified, detergent-solubilized ligand-gated ion channel, the type-3 serotonin (5-hydroxytryptamine, 5HT) receptor (5HT3R), was characterized by frequency-domain time-resolved fluorescence spectroscopy (TRFS). Detailed understanding of how ligands interact with the homopentameric receptor was obtained. While a 1:1 stoichi

While the existence of cholesterol/sphingolipid (raft) membrane domains in the plasma membrane is now supported by strong experimental evidence, the structure of these domains, their size, their dynamics, and their molecular composition remain to be understood. Raft domains are thought to represent a specific physical state of lipid bilayers, the liquid-ordered phase. Recent observations suggest that in the mammalian plasma membrane small raft domains in ordered lipid phases are in a dynamic equilibrium wit

We present a new technique for computing the electromagnetic field that propagates and is scattered in three-dimensional structures formed by bodies embedded in a stratified background. This fully vectorial technique is based on the Green's tensor associated with the stratified background. Its advantage lies in the fact that only the scatterers must be discretized, the stratified background being accounted for in the Green's tensor. Further, the boundary conditions at the different material interfaces as we

Functional immobilisation of the nicotinic acetylcholine receptor in tethered lipid membranes

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Ganglion cells from chick retina display multiple functional nAChR subtypes

Palmitoylation controls trafficking of GAD65 from Golgi membranes to axon-specific endosomes and a Rab5a-dependent pathway to presynaptic clusters

Synthesis, Testing, and Characterization of a Novel Nafion Membrane with Superior Performance in Photoassisted Immobilized Fenton Catalysis

Downscaling Fourier Transform Infrared Spectroscopy to the Micrometer and Nanogram Scale: Secondary Structure of Serotonin and Acetylcholine Receptors

Self-assembled organometallic receptors for small ions

On-line biomass monitoring with scanning dielectric spectroscopy

Study of polyelectrolyte complex formation applying the synthetic boundary technique of analytical ultracentrifugation

Accurate measurement of residual dipolar couplings in anisotropic phase

Characterization of a transmembrane receptor protein by fluorescence correlation spectroscopy

In vitro and in vivo ligand binding to the 5HT3 serotonin receptor characterised by time-resolved fluorescence spectroscopy

Raft membrane domains: from a liquid-ordered membrane phase to a site of pathogen attack

Light propagation and scattering in stratified media: a Green's tensor approach