Rates and Equilibria for a Photoisomerizable Antagonist at the Acetylcholine Receptor of Electrophorus Electroplaques

Voltage-jump and light-flash experiments have been performed on isolated Electrophorus electroplaques exposed simultaneously to nicotinic agonists and to the photoisomerizable compound 2,2'-bis-[α-(trimethylammonium)methyl]-azobenzene (2BQ). Dose-response curves are shifted to the right in a nearly parallel fashion by 2BQ, which suggests competitive antagonism; dose-ratio analyses show apparent dissociation constants of 0.3 and 1 µM for the cis and trans isomers, respectively. Flash-induced trans → cis concentration jumps produce the expected decrease in agonist-induced conductance; the time constant is several tens of milliseconds. From the concentration dependence of these rates, we conclude that the association and dissociation rate constants for the cis-2BQ-receptor binding are approximately ~ 10^8 M^(-1) s^(-1) and 60 s^(-1) at 20ºC; the Q_(10) is 3. Flash-induced cis → trans photoisomerizations produce molecular rearrangements of the ligand-receptor complex, but the resulting relaxations probably reflect the kinetics of buffered diffusion rather than of the interaction between trans-2BQ and the receptor. Antagonists seem to bind about an order of magnitude more slowly than agonists at nicotinic receptors

A covalently bound photoisomerizable agonist. Comparison with reversibly bound agonists at electrophorus electroplaques

After disulphide bonds are reduced with dithiothreitol, trans-3-(alpha-bromomethyl)-3’-[alpha-(trimethylammonium)methyl]azobenzene (trans-QBr) alkylates a sulfhydryl group on receptors. The membrane conductance induced by this “tethered agonist” shares many properties with that induced by reversible agonists. Equilibrium conductance increases as the membrane potential is made more negative; the voltage sensitivity resembles that seen with 50 [mu]M carbachol. Voltage- jump relaxations follow an exponential time-course; the rate constants are about twice as large as those seen with 50 mu M carbachol and have the same voltage and temperature sensitivity. With reversible agonists, the rate of channel opening increases with the frequency of agonist-receptor collisions: with tethered trans-Qbr, this rate depends only on intramolecular events. In comparison to the conductance induced by reversible agonists, the QBr-induced conductance is at least 10-fold less sensitive to competitive blockade by tubocurarine and roughly as sensitive to “open-channel blockade” bu QX-222. Light-flash experiments with tethered QBr resemble those with the reversible photoisomerizable agonist, 3,3’,bis-[alpha-(trimethylammonium)methyl]azobenzene (Bis-Q): the conductance is increased by cis {arrow} trans photoisomerizations and decreased by trans {arrow} cis photoisomerizations. As with Bis-Q, ligh-flash relaxations have the same rate constant as voltage-jump relaxations. Receptors with tethered trans isomer. By comparing the agonist-induced conductance with the cis/tans ratio, we conclude that each channel’s activation is determined by the configuration of a single tethered QBr molecule. The QBr-induced conductance shows slow decreases (time constant, several hundred milliseconds), which can be partially reversed by flashes. The similarities suggest that the same rate-limiting step governs the opening and closing of channels for both reversible and tethered agonists. Therefore, this step is probably not the initial encounter between agonist and receptor molecules

A photoisomerizable muscarinic antagonist. Studies of binding and of conductance relaxations in frog heart

These experiments employ the photoisomerizable compound, 3,3'-bis- [alpha-(trimethylammonium)methyl]azobenzene (Bis-Q), to study the response to muscarinic agents in frog myocardium. In homogenates from the heart, trans-Bis-Q blocks the binding of [3H]-N-methylscopolamine to muscarinic receptors. In voltage-clamped atrial trabeculae, trans- Bis-Q blocks the agonist-induced potassium conductance. The equilibrium dose-response curve for carbachol is shifted to the right, suggesting competitive blockade. Both the biochemical and electrophysiological data yield a dissociation constant of 4-5 microM for trans-Bis-Q; the cis configuration is severalfold less potent as a muscarinic blocker. Voltage-clamped preparations were exposed simultaneously to carbachol and Bis-Q and were subjected to appropriately filtered flashes (less than 1 ms duration) from a xenon flashlamp. Trans leads to cis and cis leads to trans photoisomerizations cause small (less than 20%) increases and decreases, respectively, in the agonist-induced current. The relaxation follows an S-shaped time course, including an initial delay or period of zero slope. The entire waveform is described by [1 - exp(-kt)]n. At 23 degrees C, k is approximately 3 s-1 and n is 2. Neither k nor n is affected when: (a) [Bis-Q] is varied between 5 and 100 microM; (b) [carbachol] is varied between 1 and 50 microM; (c) carbachol is replaced by other agonists (muscarine, acetylcholine, or acetyl-beta-methylcholine); or (d) the voltage is varied between the normal resting potential and a depolarization of 80 mV. However, in the range of 13-30 degrees C, k increases with temperature; the Q10 is between 2 and 2.5. In the same range, n does not change significantly. Like other investigators, we conclude that the activation kinetics of the muscarinic K+ conductance are not determined by ligand-receptor binding, but rather by a subsequent sequence of two (or more) steps with a high activation energy

Trace metal accumulation in hair and skin of the harbour seal, Phoca vitulina

Skin and hair samples of harbour seals (Phoca vitulina) found dead along the west coast of northern Germany in 1988 were analysed for total mercury, cadmium and lead. Cadmium and lead concentrations were below the detection limit in most skin samples. The mercury content of the hair (median: 23.1 μg g−1 wet wt) was several magnitudes higher than the mercury content of the skin (0.27 μg g−1 wet wt). Mercury concentrations in hair samples were significantly higher than lead concentrations (0.54 μg g−1 wet wt), which significantly exceeded cadmium levels (0.09 μg g−1 wet wt). Female seals revealed lower cadmium concentrations in the hair than male seals. Accumulations of metals with age were observed for cadmium and lead in hair samples and for mercury in skin samples of male seals. A connection between metal accumulation and pigmentation or rather moult was clearly recognizable

Editorial

Gemeenskappe se prioriteite in gesondheidsdiensThere are other human retroviruses too - a note on myelopathy associated with HTLV-IIs there a need for surgical oncologists' in South Africa

Ground state representations of loop algebras

Let g be a simple Lie algebra, Lg be the loop algebra of g. Fixing a point in S^1 and identifying the real line with the punctured circle, we consider the subalgebra Sg of Lg of rapidly decreasing elements on R. We classify the translation-invariant 2-cocycles on Sg. We show that the ground state representation of Sg is unique for each cocycle. These ground states correspond precisely to the vacuum representations of Lg.Comment: 22 pages, no figur

Classification of minimal actions of a compact Kac algebra with amenable dual

We show the uniqueness of minimal actions of a compact Kac algebra with amenable dual on the AFD factor of type II$_1$. This particularly implies the uniqueness of minimal actions of a compact group. Our main tools are a Rohlin type theorem, the 2-cohomology vanishing theorem, and the Evans-Kishimoto type intertwining argument.Comment: 68 pages, Introduction rewritten; minor correction

Fragmentable Heterogeneous Cocatalysts for the Metallocene-Catalyzed Polymerization of Olefins, I. Surface Modification of Silica and Characterization of the Resulting Carriers

Surface modification of pre-dried spherical silica nano-particles with diameters of 235 nm and 10 to 20 nm and of commercially available non-spherical silica materials using various aminoalkyltrialkoxysilanes and α-ω-bis(alkoxysilyl)organyls (organyl = alkanediyl, aminoalkanediyl, polyether, polysiloxane) has been performed in suspension in solvents with water. The quantity of water has a dominating influence, as compared to the amounts and the ratios of silanes employed, on the surface morphology of the modified silicas. The morphologies observed range from weakly linked aggregates of spherical particles to large agglomerates covered by thick irregular layers of organopolysiloxane, as demonstrated by scanning electron microscopy. These carriers can be modified further with organoaluminium compounds to yield heterogeneous cocatalysts for the polymerization of ethylene.BMBF, 03C0295, Verbundprojekt: Heterogene und homogene Cokatalysatoren und Katalysatoren für die OlefinpolymerisationDFG, GRK 352, Synthetische, mechanistische und reaktionstechnische Aspekte von Metallkatalysatore

Fragmentable Heterogeneous Cocatalysts for the Metallocene-Catalyzed Polymerization of Olefins, II [1]. Preparation, Characterization and Testing of the Cocatalysts and Microscopic Evaluation of the Polyethylene

Surface modified silica were reacted with different aluminiumalkyls AlR2R’ (R = Me, Et, i-Bu, R’ = H, Me, Et, i-Bu), oligomeric methylaluminoxane (MAO) and combinations of both, to yield heterogeneous cocatalysts. These cocatalyts were employed to polymerize ethylene using zirconocene dichloride as the catalyst. The polymerization activity profiles have been recorded and compared with the information gained from the scanning electron microscopy (SEM) images of the polymers. The fragmentation of the heterogeneous cocatalyts upon polymerization has been demonstrated. The degree of fragmentation and the polymerization activity depend on the preparation of the silica supports and on the preparation of the heterogeneous cocatalysts using these supports. The most reactive, fragmentable heterogeneous cocatalysts show polymerization activities slightly higher than MAO in homogeneous solution and almost 1.5 times higher than commercially available MAO on silica (=MAO on Sylopol).BMBF, 03C0295, Verbundprojekt: Heterogene und homogene Cokatalysatoren und Katalysatoren für die OlefinpolymerisationDFG, GRK 352, Synthetische, mechanistische und reaktionstechnische Aspekte von Metallkatalysatore