Biomedical subjects

R I Macey

Publications and source records attributed to R I Macey.

At least 37 records · Page 2Linked to original sources

PubMed1985

Apparent activation volumes of hydrophobic ions and carriers in planar lipid bilayers.

A gas-free high-pressure cell has been developed to measure planar bilayer conductances induced by hydrophobic ions and ionophores as a function of hydrostatic pressure. Plots of log conductance versus pressure for valinomycin and nonactin-mediated potassium transport in egg phosphatidyl cholinedecane membranes are essentially linear over a pressure range of 1 to 818 atm. Calculated activation volumes give similar results for both nonactin and valinomycin yielding values of + 48 and + 42 cc/mole, respectively. The valinomycin activation volume agrees reasonably well with the results obtained by Johnson and Miller (Biochim. Biophys. Acta 375:286-291, 1975) for K+-valinomycin transport in liposomes. In contrast to the activation volumes for nonactin and valinomycin, relaxation measurements of tetraphenyl boron (TPB) and dipicrylamine (DPA) give very small values of less than 5 cc/mole for the translocation rate constant, ki. Similarly, steady-state conductance measurements on tetraphenyl arsonium (TPA) and carbonylcyanide m-chlorophenylhydrazone (CCCP), give small values of 6 and 7 cc/mole, respectively. These low figures do not support transport theories based on the formation of bilayer holes or kinks (H. Träuble, J. Membrane Biol. 4:193-208, 1971). The low values for TPB and TPA are especially interesting because their cross-sectional areas are not much different than those of valinomycin and nonactin. Pressure-induced changes in membrane dielectric constant and thickness which lower the bilayer electrostatic barrier could explain the low values for the hydrophobic ions. Additionally, larger activation volumes might be expected for carriers such as nonactin and valinomycin that undergo significant rearrangement and change in hydration during surface complexation of cations.

Electric Conductivity↗

PubMed1985

Transport of hydrophobic ions in erythrocyte membrane: I. Zero membrane potential properties.

The permeability and partition coefficients of tetraphenylarsonium (TPA) and several other organic cations were studied in the human erythrocyte using an ion-selective electrode. The permeability constant for the different cations could be explained quite well by differences in oil/water partition coefficients. No evidence for facilitated transport could be found. Binding of the organic ions occurred to both the cell membrane and to intracellular contents. Partitioning to the membrane remained relatively constant despite variation from ion intracellular binding with blood samples from different donors. TPA flux is stimulated by substoichiometric amounts of tetraphenylboron and other organic anions, suggesting an ion-pairing mechanism.

Arsenicals↗

PubMed1984

Anion exchange in human erythrocytes has a large activation volume.

Sulfate equilibrium exchange in human red cells has an activation volume of +150 +/- 20 cm3/mol over the pressure range 0.1 to 83 MPa (15 to 12000 lb/in2) at 30 degrees C. This value greatly exceeds the expected contribution from sulfate binding to the anion exchanger. We suggest that the activation volume reflects conformational changes during the transport cycle.

Anion Transport Proteins↗

PubMed1984

Thermodynamics of all-or-none water channel closure in red cells.

The relation of osmotic to diffusional water permeability of human red blood cells was compared after treating the cells with different concentrations of PCMBS (p-chloromercuribenzene sulfonate). After subtracting the PCMBS-insensitive permeability (presumably the water permeability of the lipid bilayer) from each, the ratio of osmotic to diffusional permeability remains invariant (approximately equal to 11) as more and more water channels are inhibited by increasing concentrations of PCMBS. This result implies that the channels close in an all-or-none way and suggests a two-state model. Analysis of the dependence of osmotic water permeability on PCMBS concentration in terms of the model reveals a 1:1 stoichiometry and a dissociation constant for the PCMBS/membrane receptor complex of about 0.019 mM at 37 degrees C. Temperature dependence studies show that the reaction is entropically driven (delta H degrees approximately equal to 25 kcal/mol, delta S degrees approximately equal to 100 cal/mol-deg) and suggest the involvement of hydrophobic interactions.

Cell Membrane Permeability↗

PubMed1984

Transport of water and urea in red blood cells.

Evidence for water channels in red blood cells is reviewed. In an entropically driven reaction, organic mercurials decrease water permeability, elevate the activation energy, and reduce the ratio of osmotic to diffusional water permeabilities to unity so that water transport properties of red blood cells are hardly distinguishable from lipid bilayers. It is concluded that mercurials close the water channels. A variety of kinetic, pharmacological, and comparative evidence converges on the conclusion that urea and other solutes are excluded from water channels. Urea apparently permeates the red cell membrane via a facilitated diffusion system, which plays an important role when red blood cells traverse the renal medulla; rapid urea transport helps preserve the osmotic stability and deformability of the cell, and it helps prevent dissipation of extracellular osmotic gradients. Water apparently traverses the channel via a single-file mechanism; the very low channel permeability of H+ is explained if the channel contains fixed charge, or alternatively, if the mobile water molecules within the channel do not form a continuum. An alternative unitary pore hypothesis for simultaneous transport of water, ions, and small solutes is also discussed.

Biological Transport↗

PubMed1983

Determination of the electric potential at the external and internal bilayer-aqueous interfaces of the human erythrocyte membrane using spin probes.

Partitioning of oppositely charged amphipathic spin probes indicates that the electric potential at the external bilayer-aqueous interface of the human erythrocyte is insignificant, and that protruding sialic acids do not contribute to this potential. This potential at the surface is distinguished from the electrokinetic potential due to all charged groups within the hydrodynamic surface of shear. By contrast, using inside-out erythrocyte membrane vesicles, a substantial potential is observed at the cytoplasmic membrane surface. This can be attributed to the asymmetric distribution of acidic phospholipids on the two sides of the erythrocyte membrane bilayer.

Electron Spin Resonance Spectroscopy↗

PubMed1980

The permeability of the human red cell to deuterium oxide (heavy water).

Using a stopped-flow device, the osmotic water permeability of human red cells to D2O and H2O was studied as a function of temperature and under the influence of the sulfhydryl reagent paracholoromercuribenzene sulfonic acid (PCMBS), an inhibitor of water transport. The ratio, permeability (D2O)/permeability (H2O) at each temperature can be predicted simply by assuming that permeability varies inversely with macroscopic viscosity. When water permebility is inhibited with PCMBS, this dependency on viscosity vanishes; the inhibited permeabilities in D2O and H2O are indistinguishable.

Cell Membrane Permeability↗

PubMed1978

Shape and stability changes in human erythrocyte membranes induced by metal cations.

Asymmetric human erythrocyte ghost membranes behave as bilayer couple and exhibit a radius of curvature preference depending on the state of expansion or contraction of each side of the bilayer. The inside-out preference in the absence of added metal cations is gradually reduced as the K+ concentration is raised to 200 mM until a slight right-side-out preference may be exhibited Divalent cations (denoted 72+; Ca2+, Mg2+ and Mn2+) induce inside-out curvature at very low concentrations, right-side-out curvature at intermediate concentrations, and inside-out curvature again at high ones. This "triphasic" response is attributed to changes in the packing of acidic phospholipid (PL-) pairs in the A-face as a function of M2+ : PL- binding stoichiometry: 0 : 1 (PL- electrostatic repulsion and A-face expansion), 1 : 2 (PL2M crossbridging an contraction), and 1 : 1 (PLM+ repulsion and expansion). Generally increasing parent vesicle size is associated with higher cation concentrations. This is distinguished from the internal and external membrane blebbing preferred at different concentrations in accord with sidedness preference. Parent vesicle size was interpreted to be most closely associated with cation stabilisation (resistance to fragmentation) of the membrane, while sidedness and size (radius of curvature) of blebs were most closely correlated with packing of lipid molecules in the bilayer.

Adult↗

PubMed1978

Erythrocyte membrane potentials determined by hydrogen ion distribution.

If the extracellular fluid is left unbuffered, dynamic membrane potential changes in the red blood cell may be determined from external pH readings. For some types of experiments it is necessary to accelerate H+ equilibration by adding minute amounts of hydrogen carriers. The method is independent of hematocrit over a wide range of membrane potential changes. Membrane potential jumps produced by permeability changes or by changes in ionic composition may be measured. The method provides a convenient means of measuring parameters of both the conductive and non-conductive anion pathways in the red cell.

Carbonyl Cyanide m-Chlorophenyl Hydrazone↗

PubMed1977

Diffusional water permeability of red cells. Independence on osmolality.

The osmotic permeability coefficient (Lp) for human red cells has been reported to depend on the osmolality of the suspending solution. These results are also consistent with the view that the value of Lp depends on flow rectification. In this report an NMR method is used to measure the dependence of water exchange times at constant cell volume on osmolality. Our results indicate that the diffusion water permeability is constant over a large range of osmolality (300-1000 mosM) produced by the permeable solutes urea, methanol, ethanol, and glycerol. The results support the view that the apparent dependence of Lp on osmolality is due to flow rectification.

Biological Transport↗

PubMed1974