valinomycin and hydroxide-ion

This work explores the proton/hydroxide permeability (PH+/OH-) of membranes that were made of synthetic extremophile-inspired phospholipids with systematically varied structural elements. A fluorescence-based permeability assay was optimized to determine the effects on the PH+/OH- through liposome membranes with variations in the following lipid attributes: transmembrane tethering, tether length, and the presence of isoprenoid methyl groups on one or both lipid tails. All permeability assays were performed in the presence of a low concentration of valinomycin (10 nM) to prevent buildup of a membrane potential without artificially increasing the measured PH+/OH-. Surprisingly, the presence of a transmembrane tether did not impact PH+/OH- at room temperature. Among tethered lipid monolayers, PH+/OH- increased with increasing tether length if the number of carbons in the untethered acyl tail was constant. Untethered lipids with two isoprenoid methyl tails led to lower PH+/OH- values than lipids with only one or no isoprenoid tails. Molecular dynamics simulations revealed a strong positive correlation between the probability of observing water molecules in the hydrophobic core of these lipid membranes and their proton permeability. We propose that water penetration as revealed by molecular dynamics may provide a general strategy for predicting proton permeability through various lipid membranes without the need for experimentation.

Topics: Archaea; Biomimetic Materials; Fluorescent Dyes; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Hydroxides; Ionophores; Liposomes; Membrane Lipids; Membrane Potentials; Methacrylates; Microscopy, Atomic Force; Molecular Dynamics Simulation; Permeability; Protons; Unilamellar Liposomes; Valinomycin; Water

Neutral NaCl absorption is the predominant Na+ absorptive process in rat distal colon. Whether this neutral NaCl absorptive process represents Na(+)-Cl- cotransport or dual ion exchanges of Na(+)-H+ and Cl(-)-OH- has been uncertain. Recent studies using rat colonic brush-border membrane vesicles (BBMVs) have described a Na(+)-H+ exchange mechanism and have proposed that net NaCl absorption occurs via a dual ion exchange process. To date, however, an anion exchanger on the colonic apical membrane has not been identified. To determine whether a Cl(-)-OH- exchange process is present, 36Cl uptake was evaluated across rat distal colonic BBMVs. A pH gradient (7.7 in/5.5 out) stimulated active Cl- uptake. Cl- uptake was not significantly affected by the presence of a valinomycin-induced K+ diffusion potential (inside positive), suggesting that a Cl- conductive pathway is not present in these membranes. The pH gradient-stimulated Cl- uptake was a saturable function of the Cl- concentration with a Km of 14.3 +/- 5.0 mM and a Vmax of 20.4 +/- 5.5 nmol.mg protein-1.30 s-1 and was almost completely inhibited by 1 mM concentrations of SITS and DIDS, inhibitors of anion exchange processes in other epithelia. Inward gradients of Na+, K+, or Na+ and K+ did not further stimulate initial Cl- uptake, suggesting that coupling of Na+ and Cl- movement does not occur by a cotransport mechanism. Thus a Cl(-)-OH- exchange process is present in rat distal colonic BBMVs.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; 4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; Animals; Cell Membrane; Chlorides; Colon; Hydroxides; Intestinal Absorption; Kinetics; Male; Microvilli; Models, Biological; Muscle, Smooth; Rats; Rats, Inbred Strains; Sodium Chloride; Valinomycin